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1.
Rev. bras. med. esporte ; 28(5): 469-473, Set.-Oct. 2022. tab, graf
Article in English | LILACS-Express | LILACS | ID: biblio-1376709

ABSTRACT

ABSTRACT Introduction: The changes in brain structure caused by a sports-related concussion may initially be indistinguishable, however, the irreversible deleterious effects are noted in the long term. An early diagnosis may provide the patient with a better recovery chance and increased survival. For this purpose, this paper studies the feasibility of a diagnosis for concussion by microRNA (mi-RNA) biomarkers contained in the saliva of athletes. Objective: Verify whether salivary miRNAs could be considered good biomarkers for sports concussion. Methodology: Salivary mi-RNA levels were determined from 120 saliva samples of 120 players. There were 43 with a diagnosis of concussion and 77 without a diagnosis of concussion. Samples from players with a concussion were collected 30 minutes prior to activity, samples from individuals who did not engage in physical activity were also compared. Results: On the evaluation of 30 miRNA from individuals with a concussion between contact and non-contact sports there was high detection reliability(P<.05). Both miR-532-5p and miR-182-5p showed reduced amounts of physical activity. The miRNA-532-5p and miRNA-182-5p show significant results among 43 subjects from pre-exercise to post-exercise. The miRNA-4510 showed a significant result (p < 0.05) between contact and non-contact sport types. The amount of miRNA-4510 expanded in 20 individuals in the contact sport at post-exercise but remained normal in the non-contact sports group. Conclusion: The salivary miRNAs are reliable biomarkers for concussion. Evidence Level II; Therapeutic Studies - Investigating the results.


RESUMO Introdução: As alterações da estrutura cerebral provocadas por uma concussão relacionada ao esporte podem ser inicialmente indistinguíveis, porém os efeitos deletérios irreversíveis são notados a longo prazo. Um diagnóstico precoce poderá fornecer ao paciente uma chance maior de recuperação e aumento de sobrevida. Para tanto, estuda-se a viabilidade de um diagnóstico de concussão por biomarcadores de micro RNA (mi-RNA) contidos na saliva de esportistas. Objetivo: verificar se os miRNAs salivares são biomarcadores confiáveis para concussão esportiva. Metodologia: Os níveis de mi-RNA salivares foram determinados a partir de 120 amostras de saliva de 120 jogadores. Haviam 43 com diagnóstico de concussão e 77 sem diagnóstico de concussão. Amostras de jogadores com concussão foram coletadas 30 minutos antes da atividade, amostras de indivíduos que não praticaram atividade física também foram comparadas. Resultados: Na avaliação de 30 mi-RNA de indivíduos com concussão entre esportes de contato e sem contato houve grande confiabilidade de detecção(P<,05). Tanto o miR-532-5p quanto o miR-182-5p mostraram quantidades reduzidas na atividade física. O miRNA-532-5p e o miRNA-182-5p mostram resultados significativos entre 43 indivíduos desde o pré-exercício até o pós-exercício. O miRNA-4510 mostrou um resultado significativo (p < 0,05) entre os tipos de esporte com contato e sem contato. A quantidade de mi-RNA-4510 expandiu-se em 20 pessoas no esporte com contato no pós-exercício, mas permaneceu normal no grupo de esporte sem contato. Conclusão: Conclui-se que os miRNAs salivares são biomarcadores confiáveis para concussão. Nível de evidência II; Estudos Terapêuticos - Investigação de Resultados.


RESUMEN Introducción: Los cambios en la estructura cerebral causados por una conmoción encefálica relacionada con el deporte pueden ser inicialmente indistintos, sin embargo, los efectos deletéreos irreversibles se manifiestan a largo plazo. Un diagnóstico anticipado puede proporcionar al paciente una mayor posibilidad de recuperación y una mayor supervivencia. Por lo tanto, se estudia la viabilidad de un diagnóstico de conmoción encefálica mediante los biomarcadores de microARN (mi-ARN) contenidos en la saliva de los deportistas. Objetivo: verificar si los miARN salivales son biomarcadores fiables para la conmoción encefálica deportiva. Metodología: Se determinaron los niveles de mi-ARN salival a partir de 120 muestras de saliva de 120 jugadores. Había 43 con diagnóstico de conmoción encefálica y 77 sin diagnóstico de conmoción encefálica. Las muestras de los jugadores con conmoción encefálica se recogieron 30 minutos antes de la actividad, también se compararon las muestras de los individuos que no practicaban actividad física. Resultados: En la evaluación de 30 miRNA de individuos con conmoción encefálica entre deportes de contacto y sin contacto hubo una alta fiabilidad de detección (P<.05). Tanto miR-532-5p como miR-182-5p mostraron cantidades reducidas en la actividad física. El miRNA-532-5p y el miRNA-182-5p muestran resultados significativos entre los 43 sujetos desde antes del ejercicio hasta después del mismo. El miRNA-4510 mostró un resultado significativo (p < 0,05) entre los tipos de deporte de contacto y de no contacto. La cantidad de miARN-4510 se expandió en 20 individuos en el deporte de contacto en el momento posterior al ejercicio, pero se mantuvo normal en el grupo de deporte sin contacto. Conclusión: Llegamos a la conclusión de que los miARN salivales son biomarcadores fiables para la conmoción encefálica. Nivel de evidencia II; Estudios terapéuticos - Investigación de resultados.

2.
An. bras. dermatol ; 97(4): 458-466, July-Aug. 2022. tab, graf
Article in English | LILACS-Express | LILACS | ID: biblio-1383604

ABSTRACT

Abstract Background: UVB irradiation can cause acute damage such as sunburn, or photoaging and melanoma, all of which are major health threats. Objective: This study was designed to investigate the mechanism of skin photoaging induced by UVB radiation in mice through the analysis of the differential expression of miRNAs. Methods: A UVB irradiation photoaging model was constructed. HE and Masson special stains were used to examine the modifications in the epidermis and dermis of mice. The miRNA expression profiles of the mouse skin model exposed to UVB radiation and the normal skin of mice were analyzed using miRNA-sequence analysis. GO and Pathway analysis were employed for the prediction of miRNA targets. Results: A total of 23 miRNAs were evaluated for significantly different expressions in comparison to normal skin. Among them, 7 miRNAs were up-regulated and 16 were down-regulated in the skin with photoaging of mice exposed to UVB irradiation. The differential expression of miRNA is related to a variety of signal transduction pathways, among which mmu-miR-195a-5p and mitogen-activated protein kinase (MAPK) signal pathways are crucial. There was a significant differential expression of miRNA in the skin of normal mice in comparison with the skin with photoaging induced by UVB irradiation. Study limitations: Due to time and energy constraints, the specific protein level verification, MAPK pathway exploration, and miR-195a-5p downstream molecular mechanism need to be further studied in the future. Conclusions: UVB-induced skin photoaging can be diagnosed and treated using miRNA.

3.
Rev. bras. cir. cardiovasc ; 37(4): 447-453, Jul.-Aug. 2022. tab, graf
Article in English | LILACS-Express | LILACS | ID: biblio-1394729

ABSTRACT

Abstract Introduction: This study's objective is to investigate the effect of downregulation of micro ribonucleic acid (miR)-124a on myocardial injury after ischemia reperfusion (I/R) in rats. Methods: Sprague Dawley (SD) rats (n=20) were divided into four groups - sham, I/R, I/R+miR-124a antagomir (I/R+ant-miR-124a), and I/R+ant-normal control (NC). The pathomorphological and infarct size variance of injured myocardial tissues with IR were conducted with hematoxylin (HE) and triphenyltetrazolium chloride (TTC) staining. The expression levels of miR-124a, BAX, nuclear factor kappa B (NF-KB), Notch1, and Hes1 were examined by quantitative real-time polymerase chain reaction or Western blot in myocardium. The inflammatory cytokines interleukin (IL)-6, IL-1β, and tumor necrosis factor alpha (TNF-α) were detected by the enzyme-linked immunosorbent assay, as well as the activity of lactate dehydrogenase (LDH) and creatine kinase (CK) in serum by colorimetry. Results: The expression of miR-124a was increased in the I/R group. Compared with I/R and I/R+ant-NC groups, after downregulating miR-124a, the expression of IL-6, IL-1β, TNF-α, BAX, NF-KB, LDH, and CK were decreased, but the expression of Notch1 and Hes1 were increased. In HE staining, myocardial tissue edema, red blood cell exudation, and myocardial fiber arrangement disorder were accompanied by inflammatory cell infiltration and local necrosis in the I/R group. However, the pathological injury of myocardial tissue was alleviated after downregulating miR-124a. Additionally, TTC results showed that the myocardial infarction area was decreased in the I/R+ant-miR-124a group. Conclusion: Downregulation of miR-124a expression through Notch pathway can significantly reduce myocardial damage after 24 hours of I/R in SD rats. Therefore, miR-124a may become a potential therapeutic target for I/R injury.

4.
Rev. bras. cir. cardiovasc ; 37(4): 525-533, Jul.-Aug. 2022. tab, graf
Article in English | LILACS-Express | LILACS | ID: biblio-1394742

ABSTRACT

Abstract Introduction: This study investigated the correlation between the levels of long noncoding ribonucleic acids (lncRNAs) AF131217.1 and coronary slow flow (CSF). Methods: A total of 22 patients in the high-sensitivity C-reactive protein (hsCRP) group diagnosed with CSF from January 2018 to December 2018 were enrolled in this study. Coronary flow velocity was determined using the thrombolysis in myocardial infarction frame count (TFC) method. Results: LncRNA AF131217.1 expression in the CSF model was activated. Mean TFC was positively correlated with lncRNA AF131217.1 levels and hsCRP levels. LncRNA AF131217.1 induced inflammation factor levels in the in vitro model. Micro ribonucleic acid (miR)-128-3p is a target spot of lncRNA AF131217.1 on the inflammation in vitro model via Kruppel-like factor (KLF) 4. MiR-128-3p reduced inflammation factor levels (tumor necrosis factor alpha, interleukin [IL]-6, IL-1β, and IL-18). Conclusion: Thus, lncRNA AF131217.1 promoted inflammation in the regulated CSF via KLF4 by miR-128-3p.

6.
Braz. j. infect. dis ; 26(3): 102354, 2022. tab, graf
Article in English | LILACS-Express | LILACS | ID: biblio-1384132

ABSTRACT

ABSTRACT Introduction: One of the hallmarks of COVID-19 is overwhelming inflammation, which plays a very important role in the pathogenesis of COVID-19. Thus, identification of inflammatory factors that interact with the SARS-CoV-2 can be very important to control and diagnose the severity of COVID-19. The aim of this study was to investigate the expression patterns of inflammation-related non-coding RNAs (ncRNAs) including MALAT-1, NEAT-1, THRIL, and miR-155-5p from the acute phase to the recovery phase of COVID-19. Methods: Total RNA was extracted from Peripheral Blood Mononuclear Cell (PBMC) samples of 20 patients with acute COVID-19 infection and 20 healthy individuals and the expression levels of MALAT-1, NEAT-1, THRIL, and miR-155-5p were evaluated by real-time PCR assay. Besides, in order to monitor the expression pattern of selected ncRNAs from the acute phase to the recovery phase of COVID-19 disease, the levels of ncRNAs were re-measured 6-7 weeks after the acute phase. Result: The mean expression levels of MALAT-1, THRIL, and miR-155-5p were significantly increased in the acute phase of COVID-19 compared with a healthy control group. In addition, the expression levels of MALAT-1 and THRIL in the post-acute phase of COVID-19 were significantly lower than in the acute phase of COVID-19. According to the ROC curve analysis, these ncRNAs could be considered useful biomarkers for COVID-19 diagnosis and for discriminating between acute and post-acute phase of COVID-19. Discussion: Inflammation-related ncRNAs (MALAT-1, THRIL, and miR-150-5p) can act as hopeful biomarkers for the monitoring and diagnosis of COVID-19 disease.

7.
Braz. J. Pharm. Sci. (Online) ; 58: e19946, 2022. tab, graf
Article in English | LILACS | ID: biblio-1383979

ABSTRACT

Abstract The present study evaluated 56 patients diagnosed with Chronic Lymphocytic Leukemia (CLL) and a control group of 44 clinically healthy subjects with no previous history of leukemia. Genetic expressions of AKT and microRNAs were evaluated by quantitative PCR (qPCR). A significant increase in AKT gene expression in patients when compared to controls was observed (p = 0.017). When the patients were stratified according to Binet subgroups, a significant difference was observed between the subgroups, with this protein kinase appearing more expressed in the B+C subgroup (p = 0.013). Regarding miRNA expression, miR-let-7b and miR-26a were reduced in CLL patients, when compared to controls. However, no significant differences were observed in these microRNA expressions between the Binet subgroups (A versus B+C). By contrast, miR-21 to miR-27a oncogenes showed no expression difference between CLL patients and controls. AKT protein kinase is involved in the signaling cascade that occurs with BCR receptor activation, leading to increased lymphocyte survival and protection against the induction of cell death in CLL. Thus, increased AKT protein kinase expression and the reduction of miR-let-7b and miR-26a, both tumor suppressors, may explain increased lymphocyte survival in CLL patients and may be promising markers for the prognostic evaluation of this disease.


Subject(s)
Humans , Male , Female , Protein Kinases , Leukemia, Lymphocytic, Chronic, B-Cell/pathology , Patients , Gene Expression/genetics , Apoptosis , MicroRNAs/pharmacology , Healthy Volunteers
8.
Journal of Clinical Hepatology ; (12): 2091-2098, 2022.
Article in Chinese | WPRIM | ID: wpr-942665

ABSTRACT

Objective To investigate the effect of micro-ribonucleic acid-21(miR-21) on the malignant biological behavior of human cholangiocarcinoma cell line QBC939 by targeting the protein tyrosine phosphatase (PTEN)/inositol phosphate 3-kinase (PI3K)/protein kinase B (Akt) pathway. Methods The cholangiocarcinoma cell line QBC939 in the logarithmic growth phase was divided into empty vector group, blank control group, overexpression group, and silencing group.An inverted fluorescence microscope was used to observe transfection efficiency; MTT assay, flow cytometry, Transwell assay, and wound healing assay were used to measure cell proliferative activity, apoptosis rate, invasion activity, and migration activity.Quantitative reverse transcription PCR was used to measure the mRNA expression levels of miR-21, PTEN, PI3K, Akt, and mammalian target of rapamycin (mTOR); Western blotting was used to measure the protein expression levels of PTEN, PI3K, Akt, phosphorylated Akt (p-Akt), and mTOR; dual-luciferase reporter assay was used to verify the effect of miR-21 on PTEN.A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the SNK- q test was used for further comparison between two groups. Results Transfection efficiency was 90.27%±18.03% in the overexpression group, 91.43%±18.26% in the silencing group, and 92.16%±18.41% in the empty vector group.Compared with the blank control group and the empty vector group, the overexpression group had a significant increase in the proliferative activity of QBC939 cells (both P < 0.05) and a significant reduction in apoptosis rate (both P < 0.01);compared with the blank control group, the empty vector group, and the overexpression group, the silencing group had a significant reduction in proliferative activity ( P < 0.01) and a significant increase in apoptosis rate ( P < 0.01).Compared with the blank control group and the empty vector group, the overexpression group had significant increases in the migration rate of QBC939 cells and number of cells penetrating the membrane (all P < 0.01);compared with the blank control group, the empty vector group, and the overexpression group, the silencing group had significant reductions in migration rate and number of cells penetrating the membrane (all P < 0.01).Compared with the blank control group and the empty vector group, the overexpression group had significant increases in the mRNA expression levels of miR-21, PI3K, Akt, and mTOR and a significant reduction in the mRNA expression level of PTEN (all P < 0.05);compared with the blank control group, the empty vector group, and the overexpression group, the silencing group had significant reductions in the mRNA expression levels of miR-21, PI3K, Akt, and mTOR and a significant increase in the mRNA expression level of PTEN (all P < 0.05).Compared with the blank control group and the empty vector group, the overexpression group had significant increases in the protein expression levels of PI3K, Akt, p-Akt, and mTOR and a significant reduction in the protein expression level of PTEN (all P < 0.01);compared with the blank control group, the empty vector group, and the overexpression group, the silencing group had significant reductions in the protein expression levels of PI3K, Akt, p-Akt, and mTOR and a significant increase in the protein expression level of PTEN (all P < 0.01).Furthermore, miR-21 showed targeted regulation of PTEN expression. Conclusion MiR-21 silencing may inhibit the malignant biological behavior of human cholangiocarcinoma cell line QBC939 by targeting the PTEN/PI3K/Akt signaling pathway to upregulate the expression of PTEN and downregulate the expression of PI3K, Akt, mTOR, and p-Akt.

9.
Article in English | WPRIM | ID: wpr-939825

ABSTRACT

Mammalian bone is constantly metabolized from the embryonic stage, and the maintenance of bone health depends on the dynamic balance between bone resorption and bone formation, mediated by osteoclasts and osteoblasts. It is widely recognized that circadian clock genes can regulate bone metabolism. In recent years, the regulation of bone metabolism by non-coding RNAs has become a hotspot of research. MicroRNAs can participate in bone catabolism and anabolism by targeting key factors related to bone metabolism, including circadian clock genes. However, research in this field has been conducted only in recent years and the mechanisms involved are not yet well established. Recent studies have focused on how to target circadian clock genes to treat some diseases, such as autoimmune diseases, but few have focused on the co-regulation of circadian clock genes and microRNAs in bone metabolic diseases. Therefore, in this paper we review the progress of research on the co-regulation of bone metabolism by circadian clock genes and microRNAs, aiming to provide new ideas for the prevention and treatment of bone metabolic diseases such as osteoporosis.


Subject(s)
Animals , Circadian Clocks/genetics , Circadian Rhythm/genetics , Mammals/genetics , MicroRNAs/genetics , Osteogenesis/genetics , Osteoporosis/genetics
10.
Article in Chinese | WPRIM | ID: wpr-939510

ABSTRACT

OBJECTIVE@#To observe the effect of moxibustion on the regulation of nuclear factor-kappa B (NF-κB) and inflammatory factors by multiple microRNAs (miRNAs) in rats with diarrhea-predominant irritable bowel syndrome (IBS-D), and to explore the anti-inflammatory mechanism of moxibustion on IBS-D.@*METHODS@#Twelve of 52 newborn rats were randomly selected into a normal group. The remaining rats were made into IBS-D model. A total of 36 rats with successful model were randomly divided into a model group, a medication group and a moxibustion group, 12 rats in each group. The rats in the medication group were intraperitoneally injected with pyrrolidine dithiocarbamate (PDTC). The rats in the moxibustion group were treated with moxibustion at "Tianshu" (ST 25) and "Shangjuxu" (ST 37) for 20 min each time. All the intervention was given once a day for 7 days. Before and after modeling as well as after intervention, the body mass, loose stool rate and the minimum volume threshold of abdominal withdrawal reflex (AWR) were measured. After intervention, the contents of serum tumor necrosis factor α (TNF-α), interleukin (IL)-1β and IL-8 were detected by ELISA method; the morphology of colon tissues was observed by HE staining, and the expressions of miR-155, miR-125b, miR-29b, miR-31, miR-18a and NF-κB p65 mRNA in colon tissues were detected by real-time PCR. The expressions of NF-κB p65, TNF-α, IL-1β and IL-8 protein in colon tissues were detected by immunofluorescence.@*RESULTS@#After modeling, the body mass and the minimum volume threshold of AWR in the model group were lower than those in the normal group (P<0.01); the rates of loose stool in the model group were higher than those in the normal group (P<0.01); after intervention, in the model group, the inflammatory infiltration of colon tissues was obvious, and the serum levels of TNF-α, IL-1 β, IL-8 were higher than those in the normal group (P<0.05); the expression of miR-155, miR-125b, miR-29b, miR-31, miR-18a and NF-κB p65 mRNA in colon tissues was higher than that in the normal group (P<0.05); the protein expression of NF-κB p65, TNF-α, IL-1β, IL-8 was also higher than that in the normal group (P<0.01). After intervention, the body mass and the minimum volume threshold of AWR in the medication group and the moxibustion group were both higher than those in the model group (P<0.05); the loose stool rate in the medication group and the moxibustion group were lower than those in model group (P<0.05); the inflammatory cells infiltration in the colon tissues was less, the serum levels of TNF-α, IL-1β and IL-8 as well as the protein expression of NF-κB p65, TNF-α, IL-1β and IL-8 in the colon tissues in the medication group and the moxibustion group were lower than those in the model group (P<0.05, P<0.01). The expression of miR-125b, miR-31, miR-18a and NF-κB p65 mRNA in the medication group were lower than those in the model group (P<0.05). The expression of miR-155, miR-125b, miR-29b, miR-31, miR-18a and NF-κB p65 mRNA in the moxibustion group were lower than those in the model group (P<0.05). The miR-155, miR-125b, miR-29b, miR-31, miR-18a were positively correlated with NF-κB p65 mRNA (0<r<1, P<0.01).@*CONCLUSION@#The anti-inflammatory mechanism of moxibustion at "Tianshu" (ST 25) and "Shangjuxu" (ST 37) for IBS-D rats may be related to regulating multiple miRNAs to inhibit NF-κB signal pathway and reduce the expression of inflammatory factors.


Subject(s)
Animals , Diarrhea/therapy , Interleukin-8/genetics , Irritable Bowel Syndrome/therapy , MicroRNAs/genetics , Moxibustion , NF-kappa B/metabolism , RNA, Messenger , Rats , Rats, Sprague-Dawley , Signal Transduction , Tumor Necrosis Factor-alpha/genetics
11.
Article in Chinese | WPRIM | ID: wpr-935802

ABSTRACT

Arsenic is a non-metallic element, and the International Agency for Research on Cancer has identified arsenic and its compounds as carcinogens. Arsenic and its compounds can be absorbed through the respiratory tract, skin and digestive tract, distributed in the liver, kidney, lung and skin, and cause damage. Non-coding RNAs are closely related to arsenic-induced nervous system disorders, cell necrosis, reproductive toxicity, and carcinogenesis. In recent years, the network regulation of microRNAs (miRNAs) , long non-coding RNAs (lncRNAs) , and circular RNAs (circRNAs) among non-coding RNAs in various diseases induced by arsenic has become a new research field. This paper summarizes the existing scientific research results, and expounds the mechanism of miRNAs, lncRNAs and circRNAs in arsenic toxicity, and provides basic data and theoretical basis for the prevention and treatment of arsenic poisoning.


Subject(s)
Arsenic/toxicity , Arsenic Poisoning , Humans , MicroRNAs/genetics , RNA, Circular , RNA, Long Noncoding/genetics
12.
Cancer Research and Clinic ; (6): 171-175, 2022.
Article in Chinese | WPRIM | ID: wpr-934651

ABSTRACT

Objective:To explore the effects of long non-coding RNA (lncRNA) FAM224A on the proliferation and migration of ovarian cancer cells by regulating the expression of miRNA-590-3p (miR-590-3p).Methods:Human ovarian cancer cell lines OC3, SKOV-3, HO-8910, A2780 and human normal ovarian epithelial cell line IOSE80 were selected, and the relative expression of FAM224A in each cell line was detected by real-time quantitative polymerase chain reaction (qRT-PCR). The cell line with the lowest relative expression level of FAM224A was screened for follow-up experiment. The cells were divided into FAM224A group (transfected with FAM224A mimic gene) and control group (transfected with control mimic gene). CCK-8 method and cell scratch test were used to detect the cell proliferation and migration ability of the two groups. The bioinformatics website LncBase v.2 predicted that the target gene that FAM224A might complementarily bind to was miR-590-3p. qRT-PCR was used to detect the relative expression levels of miR-590-3p and forkhead box protein A2 (FOXA2) mRNA, and the expressions of related proteins were detected by Western blot.Results:The relative expression levels of FAM224A in ovarian cancer cell lines OC3, SKOV-3, HO-8910, A2780 and normal ovarian epithelial cell line IOSE80 were 0.23±0.04, 0.65±0.05, 0.45±0.03, 0.63±0.08 and 1.02±0.11, respectively, and the difference was statistically significant ( F = 14.78, P < 0.01), and the cell line with the lowest relative expression level of FAM224A was OC3. The results of CCK-8 method showed that the proliferation ability of OC3 cells in the FAM224A group was lower than that in the control group on the 2nd, 3rd, 4th and 5th day of culture (all P < 0.05). The scratch healing rates of OC3 cells in the FAM224A group and the control group were (18.6±2.3)% and (71.7±7.2)%, respectively, and the difference was statistically significant ( t = 6.99, P < 0.01). The relative expression levels of FAM224A in OC3 cells in the FAM224A group and the control group was 12.36±1.45 and 1.14±0.24, respectively ( t = 13.08, P < 0.01); the relative expression levels of miR-590-3p were 0.19±0.06 and 1.04±0.20, respectively ( t = 4.01, P < 0.01); the relative expression levels of FOXA2 mRNA were 6.37±1.37 and 1.05±0.08, respectively ( t = 3.86, P < 0.01). Compared with the control group, the expression of FOXA2 protein in OC3 cells in the FAM224A group was increased, the expressions of cell proliferation protein cyclin-dependent kinase 2 (CDK2) and cyclin D3 were decreased, and the expression of cell migration protein Snail was decreased. Conclusions:FAM224A is low expressed in ovarian cancer cell lines. FAM224A reduces the proliferation and migration ability of ovarian cancer OC3 cells by inhibiting the expression of miR-590-3p.

13.
Cancer Research and Clinic ; (6): 161-165, 2022.
Article in Chinese | WPRIM | ID: wpr-934649

ABSTRACT

Objective:To explore the effects of scutellarein on the cell proliferation and migration of cervical cancer cell line HCC94 through miRNA-496 (miR-496) and stromal cell-derived factor 1 (SDF-1).Methods:HCC94 cells in the logarithmic growth phase were taken as objects, and 20 μmol/L scutellarin solution (scutellarin group) and dimethyl sulfoxide (DMSO) (control group) were added respectively. The CCK-8 method and Transwell experiment were used to detect the proliferation and migration ability of the two groups of HCC94 cells. The relative expression levels of miR-496 and SDF-1 mRNA were detected by real-time quantitative polymerase chain reaction (qRT-PCR). The target gene of miR-496 was predicted by the bioinformatics software TarBase and verified by the dual-luciferase reporter gene assay. Western blot was used to detect the expression of related proteins.Results:Compared with the control group, the proliferation ability of HCC94 cells in the scutellarin group was decreased on the 3rd, 4th and 5th day of culture (all P < 0.05). The number of HCC94 cells in the scutellarin group and the control group were 25±5 and 134±19, respectively, and the cell migration ability of the scutellarin group was lower than that of the control group ( t = 5.61, P < 0.01). The relative expressions of miR-496 in the control group and the scutellarin group were 1.07±0.12 and 11.24±2.75, respectively, and the difference was statistically significant ( t = 3.68, P < 0.01). The dual-luciferase reporter gene assay confirmed that SDF-1 was the downstream targeted gene of miR-496. The relative expressions of SDF-1 mRNA in the scutellarin group and the control group were 0.29±0.05 and 1.01±0.07, respectively, and the difference was statistically significant ( t = 7.22, P < 0.01). Compared with the control group, after scutellarin promoted the expression of miR-496, the expressions of SDF-1 protein, the cell proliferation protein cyclin-dependent kinase 3 (CDK3) and the cell migration proteins Slug and Zeb-2 were decreased. Conclusions:Scutellarin could inhibit the proliferation and migration of cervical cancer HCC94 cells through the miR-496-SDF-1 axis.

14.
Cancer Research and Clinic ; (6): 86-91, 2022.
Article in Chinese | WPRIM | ID: wpr-934634

ABSTRACT

Objective:To explore the effects of miRNA-373-3p (miR-373-3p) on the proliferation of nephroblastoma G401 cells through targeted regulation of CD44 expression.Methods:Bioinformatic method was used to predict the possible targeted genes of miR-373-3p based on bioinformatic databases including miRDB, miRanda, PITA and DIANA-microT. G401 cells were taken and transfected with miR-373-3p mimic, mimic negative control, miR-373-3p inhibitor or inhibitor negative control, respectively. Cell proliferation ability was detected by using CCK-8 assay. The number of clones was detected by using clone formation assay. The relative expression level of CD44 mRNA was detected by using real-time fluorescent quantitative polymerase chain reaction (qRT-PCR), and the expression level of CD44 protein was detected by using Western blotting. The dual luciferase gene reporter assay was carried out in HEK-293T cells to vertify the target gene of miR-373-3p.Results:Bioinformatic analysis indicated that CD44 was a targeted gene of miR-373-3p. After 24 h transfection, the proliferation activity of G401 cells in miR-373-3p mimic group was decreased compared with that in mimic negative control group (all P < 0.05). After 48 h transfection, the proliferation activity of tumor cells in miR-373-3p inhibitor group was increased compared with that inhibitor negative control group (all P < 0.05). The formed number of clones in miR-373-3p mimic group was reduced compared with that in the mimic negative control group (55.3±2.5 vs. 90.7±2.9), and the difference was statistically significant ( t = 14.57, P < 0.01). The formed number of clones in miR-373-3p inhibitor group was more than that in inhibitor negative control group (115.0±2.7 vs. 92.0±2.4), and the difference was statistically significant ( t = 8.86, P < 0.01). The dual-luciferase gene reporter assay showed that CD44 was a direct targeted gene of miR-373-3p. The relative expression levels of CD44 mRNA in miR-373-3P mimic and mimic negative control group were 0.62±0.03 and 1.00±0.01, respectively, and the difference was statistically significant ( t = 11.28, P < 0.01). The relative expression levels of CD44 mRNA in miR-373-3p inhibitor and inhibitor negative control group were 1.31±0.02 and 1.00±0.00, respectively, and the difference was statistically significant ( t = 12.65, P < 0.01). The CD44 protein expression was decreased in miR-373-3p mimic group, while increased in miR-373-3p inhibitor group. Conclusion:miR-373-3p can inhibit tumor cell proliferation by targeting CD44 in nephroblastoma.

15.
Cancer Research and Clinic ; (6): 81-85, 2022.
Article in Chinese | WPRIM | ID: wpr-934633

ABSTRACT

Objective:To investigate the effects of astragalin on the cell proliferation and cell cycle of prostate cancer cell line C4-2B through up-regulating the expression of miRNA-513 (miR-513).Methods:Prostate cancer cell line C4-2B cells were taken and treated with 125 μg/L of astragalin for 48 h (astragalin group), and untreated C4-2B cells were set as the control group. The methyl thiazolyl tetrazolium (MTT) method was used to detect the proliferation ability of C4-2B cells in the two groups, and cell cycle was detected by using flow cytometry. The miRNAMap prediction software was used to predict that the targeted gene of miR-513 was the forkhead box protein R2 (FOXR2), and the dual luciferase gene reporter assay was used to verify it. Real-time fluorescent quantitative polymerase chain reaction (qRT-PCR) was used to detect the relative expression levels of miR-513 and FOXR2 mRNA in the two groups of cells. Western blotting was used to detect the expressions of FOXR2, cyclin-dependent kinase 7 (CDK7), β-actin and cyclin H in the two groups of C4-2B cells.Results:Compared with the control group, the proliferation activity of C4-2B cells in the astragalin group was decreased from day 2 to day 5 (all P < 0.05). The proportions of S-phase cells in the control group and the astragalin group were (48.1±3.2)% and (36.0±2.1)%, respectively. The proportion of S-phase cells in the astragalin group was decreased ( t = 3.12, P = 0.021); the proportions of G 2-phase cells were (24.9±3.3)% and (11.8±2.4)%, respectively. The proportion of G 2-phase cells in the astragalin group was decreased ( t = 3.18, P = 0.019). The relative expression levels of miR-513 in C4-2B cells of the control group and the astragalin group were 1.01±0.22 and 6.55±0.61, respectively. The relative expression levels of miR-513 in C4-2B cells in the astragalin group was increased ( t = 7.70, P < 0.01). The dual luciferase reporter gene assay verified that FOXR2 was the targeted gene of miR-513. The relative expression level of FOXR2 mRNA in C4-2B cells of the control group and the astragalin group was 1.04±0.14 and 0.19±0.06, respectively, and the difference was statistically significant ( t = 5.53, P = 0.002), suggesting that after astragalin promoted the expression of miR-513, the FOXR2 mRNA expression was decreased. The relative expression levels of FOXR2, CDK7 and cyclin H protein in C4-2B cells in the astragalin group were all decreased compared with those in the control group. Conclusions:Astragalin inhibits the proliferation of prostate cancer C4-2B cells and induces cell cycle arrest by up-regulating the expression of miR-513.

16.
Cancer Research and Clinic ; (6): 51-55, 2022.
Article in Chinese | WPRIM | ID: wpr-934627

ABSTRACT

Objective:To investigate the effects of serum miRNA-126 (miR-126) and miRNA-449a (miR-449a) expression levels on the efficacy and prognostic evaluation of pemetrexed combined with cisplatin in treatment of non-small cell lung cancer (NSCLC).Methods:The data of 100 NSCLC patients admitted to Wuhan Hankou Hospital from January 2016 to January 2017 were retrospectively analyzed. Before treatment, the relative expression levels of miR-126 and miR-449a in serum of both groups were detected by using real-time fluorescence quantitative polymerase chain reaction. A total of 100 healthy volunteers during the same period were treated as the control group. The efficacy was evaluated after 2 courses of treatment, and the correlation of miR-126 and miR-449a relative expression levels with clinicopathological characteristics, efficacy and prognosis of patients was analyzed.Results:The relative expression levels of serum miR-126 and miR-449a in NSCLC patients were lower than those in the control group (1.23±0.34 vs. 2.22±0.57, 0.95±0.21 vs. 2.13±0.43), and the differences were statistically significant ( F = 14.92, 24.66; all P < 0.01). There were significant differences in the relative expression levels of serum miR-126 and miR-449a in NSCLC patients with different therapeutic effects ( F = 80.65, 22.43, all P < 0.01). Spearman correlation analysis showed that the relative expression levels of serum miR-126 and miR-449a were correlated with the therapeutic effect ( ρ = 0.782, 0.618, all P < 0.01). The relative expression levels of serum miR-126 and miR-449a in NSCLC patients with different gender and age had no statistically significant difference (all P > 0.05). The relative expression levels of serum miR-126 and miR-449a in NSCLC patients with different TNM staging, pathological type, tumor diameter, degree of differentiation showed significant differences (all P < 0.05). Univariate Cox regression analysis showed TNM staging, tumor diameter, degree of differentiation, relative expression levels of serum miR-126 and miR-449a were correlated with the survival of NSCLC patients (all P < 0.05). Multivariate Cox regression analysis revealed TNM staging, tumor diameter, degree of differentiation and the relative expression levels of miR-126 and miR-449a were independent factors affecting the survival of NSCLC patients after surgery. The overall survival of patients with the high expressions of miR-126 and miR-449a was better than that of patients with the low expressions (all P < 0.05). Conclusions:The relative expression levels of serum miR-126 and miR-449a can be used as a potential serologic indicator to analyze the prognosis of NSCLC patients treated with pemetrexed combined with cisplatin.

17.
Cancer Research and Clinic ; (6): 19-25, 2022.
Article in Chinese | WPRIM | ID: wpr-934621

ABSTRACT

Objective:To investigate the effect of miRNA-296-5p (miR-296-5p) on the migration and invasion of hypoxia-induced pancreatic cancer cells and its related mechanisms.Methods:Human pancreatic cancer cell line PANC-1 was selected. Pancreatic cancer tissues from 55 pancreatic cancer patients who underwent the resection and adjacent carcinoma normal pancreatic tissues from 10 patients at Shanghai Fengxian District Central Hospital and Bengbu Medical College First Affiliated Hospital between January 2010 and December 2014 were collected. The expression levels of hypoxia-inducible factor 1α (HIF-1α) and miR-296-5p in tissue microarray of pancreatic cancer and adjacent carcinoma normal pancreatic tissues were detected by using immunohistochemistry and in situ hybridization. The relationship between miR-296-5p and HIF-1α as well as their correlation with clinicopathological characteristics of patients were analyzed. PANC-1 cells were divided into hypoxic group and normoxic group. Transwell assay was used to detect the cell migration and invasion ability of both groups. Real-time quantitative polymerase chain reaction (qRT-PCR) was used to examine the expressions of HIF-1α and miR-296-5p under hypoxic environment of both groups. The expression of HIF-1α was interfered by transfecting small interfering RNA (siRNA). PANC-1 cells were divided into PANC-1 group (the empty control), PANC-1-NC group (the negative control) and PANC-1-siRNA group. The expression of miR-296-5p was measured. After co-transfecting miR-296-5p agonist and miR-296-5p inhibitor, the cells were divided into Agomir-miR-296-5p group (agonist group), Agomir-miR-296-5p-NC group (agonist negative control group), Antagomir-miR-296-5p group (inhibitor group) and Antagomir-miR-296-5p-NC group (inhibitor negative control group). Transwell assay was used to detect the cell migration and invasion ability of all groups. Luciferase reporter gene system was used to verify whether miR-296-5p promoter region had binding site of HIF-1α.Results:The high expression rate of HIF-1α in pancreatic cancer tissues was higher than that of adjacent carcinoma normal pancreatic tissues [81.8% (45/55) vs. 0 (0/10), P<0.01], and the high expression rate of miR-296-5p in pancreatic cancer tissues was lower than that of adjacent carcinoma normal pancreatic tissues [12.7% (7/55) vs. 90.0% (9/10), χ2 = 27.23, P<0.01]. The expression of HIF-1α was negatively correlated with that of miR-296-5p ( r = -0.53, P<0.01). The low expression of miR-296-5p was closely related with the tumor diameter, TNM staging, lymph node metastasis (all P<0.05). The number of PANC-1 invasion cell was 15.3±2.1 in normoxic group and 24.7±1.5 in hypoxic group, and the difference was statistically significant ( t = 0.26, P = 0.003). The number of PANC-1 migration cell was 20.7±3.8 in hypoxic group and 32.7±1.2 in normoxic group, and the difference was statistically significant ( t = 5.25, P = 0.006). The relative expression level of HIF-1α mRNA in PANC-1 cell of hypoxic group was higher than that of normoxic group [(1.00±0.01) vs. (0.30±0.02)], and the difference was statistically significant ( t = 56.45, P<0.01); the relative expression level of miR-296-5p in PANC-1 cell of hypoxic group was lower than that of normoxic group [(1.14±0.04) vs. (3.05±0.20)], and the difference was statistically significant ( t = 16.05, P<0.01). The number of invasion cells in PANC-1 group, PANC-1-NC group and PANC-1-siRNA group was 24.7±1.5, 25.7±1.5, 12.0±1.7, respectively, and the difference was statistically significant ( F = 68.13, P<0.01).The cell invasion ability in PANC-1-siRNA group was decreased compared with that in PANC-1 group ( t = 9.50, P = 0.001). The number of cell migration was 32.7±1.2, 37±1.0, 17.3±1.2, respectively in PANC-1 group, PANC-1-NC group and PANC-1-siRNA group, and the difference was statistically significant ( F = 262.09, P<0.01). The cell migration ability in PANC-1-siRNA group was decreased compared with that in PANC-1 group ( t = 16.26, P<0.01). The cell invasion and migration ability in Antagomir-miR-296-5p group was increased compared with that in PANC-1 group (all P<0.05); the cell invasion and migration ability in Agomir-miR-296-5p group was decreased compared with that in PANC-1 group (all P<0.05). The results of luciferase activity detected by luciferase reporter gene system showed that miR-296-5p had the target binding to HIF-1α. Conclusions:HIF-1α plays a key role in the invasion and migration of hypoxia-induced pancreatic cancer cells through negatively reducing miR-296-5p.

18.
Article in Chinese | WPRIM | ID: wpr-934272

ABSTRACT

Objective:To observe the inhibitory effect of lentivirus (LV)-mediated miR-191 on the proliferation and angiogenesis of human retinal vascular endothelial cells (hREC) cultured in vitro.Methods:The hREC cell lines were cultured in vitro and divided into control group, hypoxia group, LV-empty vector (LV-vector) group, and LV-miR-191 (LV-191) group. The LV-vector group and LV-191 group were transferred to the corresponding lentiviral vector respectively. Flow cytometry was used to detect cell transfection efficiency. Cell Counting Kit-8 (CCK-8) test was used to detect cell proliferation ability. Scarification test and invasion chamber (Transwell) test were used to detect cell migration ability. Matrigel test was used to detect cell lumen formation ability. Real-time quantitative polymerase chain reaction (qPCR) was used to detect the relative expression of miR-191 and relative mRNA expression of its downstream target genes p21, vascular endothelial growth factor (VEGF), cell division protein kinase (CDK) 6, cyclin-D1 (Cyclin D1). Independent sample t test was used for pairwise comparison. Results:The results of flow cytometry showed that the transfection efficiency of cells in the control group and the LV-191 group were 0.615% and 99.400%, respectively. The results of CCK-8, scarification, Transwell and Matrigel test showed that, compared with the control group, the number of cell proliferation ( t=6.130, 4.606), the cell mobility ( t=4.910, 6.702), the number of stained cells on the microporous membrane ( t=7.244, 6.724) and the lumen formation ability cells ( t=8.345, 9.859) were significantly increased in the hypoxia group and the LV-vector group ( P<0.01), while the LV-191 group showed completely opposite performance ( t=14.710, 6.245, 5.333, 5.892; P≤0.01). The qPCR test results showed that, compared with the control group and the LV-vector group, the relative expression of miR-191 mRNA in the cells of the LV-191 group was significantly up-regulated ( t=44.110, 42.680), the relative expression of Cyclin D1 mRNA ( t=29.940, 14.010) and CDK6 mRNA ( t=15.200, 7.645) decreased significantly, and the difference were statistically significant ( P<0.01); the relative expression of p21 mRNA increased, however, the difference was not statistically significant ( t=2.013, 2.755; P>0.05). There was no significant difference in the relative expression of VEGF mRNA in the 4 groups of cells ( F=0.966, P>0.05). Conclusions:LV-191 can inhibit the proliferation, migration and tubing of hREC by up-regulating p21 and down-regulating CDK6 and Cyclin D1.

19.
Chinese Journal of Nephrology ; (12): 336-343, 2022.
Article in Chinese | WPRIM | ID: wpr-933865

ABSTRACT

Objective:To investigate the protective effect and potential mechanisms of microRNA-26a-5p (miR-26a-5p) on podocyte injury in diabetic kidney disease (DKD).Methods:(1) In vivo experiment: Four-week-old db/db mice were divided into db/db group, db/db+agomir-NC group and db/db+miR-26a-5p agomir group according to random number table method, with 10 mice in each group, and 10 db/m mice of the same week-old were set as normal control group. At the age of 10 weeks, pathological changes were observed through light and electron microscopy. Kidney weight/body weight (KW/BW), urinary albumin to creatinine ratio (ACR), fasting blood glucose (FBG) and other biochemical indicators were also detected. The position and expression of miR-26a-5p in kidney tissue were determined through fluorescence in situ hybridization and quantitative real-time PCR, while the expressions of transient receptor potential cation channel-6 (TRPC6) and Nephrin in kidney tissue were determined by Western blotting and immunohistochemistry. (2) In vitro experiment: The immortalized mouse podocytes (MPC5) were divided into 5 groups: normal glucose group, high mannitol group, high glucose group, high glucose+miR-26a-5p mimic group, and high glucose+mimic-NC group. The expressions of miR-26a-5p, TRPC6 and Nephrin were detected. Luciferase reporter assay was conducted to research the relationship of miR-26a-5p and TRPC6. Results:(1) In vivo experiment: Compared with db/m group, db/db mice exhibited lower KW/BW and disrupted conditions of ACR, FBG, total cholesterol, triglycerides and low density lipoprotein cholesterol (all P<0.01). Increased glomeruli volume, more extracellular matrix deposition, thicker basement membrane and more foot process fusion were observed by light and electron microscope. Increased expression of TRPC6 protein as well as decreased expression of Nephrin protein and miR-26a-5p were detected in kidney tissues of db/db mice ( P<0.05). Compared with db/db+agomir-NC group, db/db mice transfected by miR-26a-5p agomir exhibited less albuminuria, with less protein expression of TRPC6 and more Nephrin in kidney tissue (all P<0.05). (2) In vitro experiment: Compared with normal glucose group, high glucose-treated podocytes exhibited increased expression of TRPC6 ( P<0.05), as well as decreased expression of Nephrin ( P<0.05) and miR-26a-5p ( P<0.01). Compared with high glucose+mimic-NC group, lower expression of TRPC6 and higher expression of Nephrin were detected in podocytes transfected by miR-26a-5p mimic (both P<0.05). Luciferase reporter assay confirmed that miR-26a-5p could regulate the expression of TRPC6 precisely. Conclusions:The expression of miR-26a-5p in podocytes is down-regulated in the context of high glucose and miR-26a-5p protects podocytes from injury via inhibiting the expression of TRPC6 in DKD.

20.
Article in Chinese | WPRIM | ID: wpr-933302

ABSTRACT

Objective:To evaluate the role of has_circ_0008039 and miR-484 in oxygen-glucose deprivation/reoxygenation (OGD/R) injury in SK-N-SH cells and the relationship with Fis1.Methods:SK-N-SH cells were cultured in vitro to logarithmic growth stage and divided into 5 groups ( n=25 each) according to the random number table method: control group (group C), OGD/R group, has_circ_0008039 siRNA group (group S), hsa_circ_0008039 over-expression group (group E) and has_circ_0008039 siRNA plus miR-484 inhibitor group (group S+ I). Cells were cultured in normal condition in group C. In S, E and S+ I groups, after the cells were transfected with hsa_circ_0008039 siRNA, has_circ_0008039 over-expression vector, hsa_circ_0008039 siRNA and miR-484 inhibitor, the cells were subjected to oxygen-glucose deprivation for 12 h followed by 24 h restoration of O 2-glucose supply to develop the OGD/R model.At 24 h of restoration of O 2-glucose supply, the cell viability and amount of lactic dehydrogenase (LDH) released were measured using CCK-8 assay, the expression of hsa_circ_0008039, miR-484 and Fis1 mRNA was detected using real-time polymerase chain reaction, and the expression of Fis1 was detected by Western blot.A dual-fluorescein experimental report was used to verify the targeting relationship between hsa_circ_0008039 and miR-484. Results:Compared with group C, the cell viability was significantly decreased, and the amount of LDH released was increased in the other 4 groups, the expression of hsa_circ_0008039 and Fis1 was significantly up-regulated, and the expression of miR-484 was down-regulated in OGD/R and E groups, the expression of hsa_circ_0008039 and Fis1 was significantly down-regulated, and miR-484 was up-regulated in group S, and the expression of hsa_circ_0008039 and miR-484 was significantly down-regulated, and the expression of Fis1 was up-regulated in group S+ I ( P<0.05). Compared with group OGD/R, the cell viability was significantly decreased, and the amount of LDH released was increased in E and S+ I groups, the cell viability was significantly increased, and the amount of LDH released was decreased in group S, the expression of hsa_circ_0008039 and Fis1 was significantly up-regulated, and the expression of miR-484 was down-regulated in group E, the expression of hsa_circ_0008039 and Fis1 was significantly down-regulated, and the expression of miR-484 was up-regulated in group S, and the expression of hsa_circ_0008039 and miR-484 was significantly down-regulated, and the expression of Fis1 was up-regulated in group S+ I ( P<0.05). Compared with group S, the cell viability was significantly decreased, the amount of LDH released was increased, the expression of miR-484 was down-regulated, and the expression of Fis1 was up-regulated in group S+ I ( P<0.01). The dual-fluorescein experimental report verified that miR-484 was the target of hsa_circ_0008039 which binded to miR-484 specifically. Conclusions:has_circ_0008039 is involved in OGD/R injury in SK-N-SH cells by targetedly binding to miR-484, which is associated with up-regulation of Fis1 expression.

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