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1.
Article in Chinese | WPRIM | ID: wpr-1027467

ABSTRACT

Objective:To investigate the relationship between serum miR-501 and miR-195 levels and sensitivity to concurrent chemoradiotherapy in patients with locally advanced cervical cancer (LACC).Methods:Clinical data of 96 patients with LACC admitted to Nanyang Central Hospital from January 2020 to June 2022 were retrospectively analyzed and compared with those of 96 healthy subjects during physical examination in our hospital during the same period to compare the differences of serum miR-501 and miR-195 levels. Tumor status was reviewed at 6 months after concurrent chemoradiotherapy. Patients were divided into the sensitive and resistant groups according to the evaluation criteria of solid tumor efficacy. The relationship between serum miR-501 and miR-195 levels and the sensitivity to concurrent chemoradiotherapy in LACC patients was analyzed by univariate and multivariate analyses. The receiver operating characteristic (ROC) curve was also drawn to predict differential efficacy of concurrent chemoradiotherapy sensitivity in LACC patients. Multivariate analysis was conducted by binary logistic regression analysis. P<0.05 indicated statistically significant differences. Results:In LACC patients, serum miR-501 level was significantly higher, whereas serum miR-195 level was significantly lower than those in physical examination subjects (both P<0.05). Univariate analysis showed that serum miR-501 level at admission in the resistant group was significantly higher, whereas serum miR-195 level was significantly lower than those in the sensitive group (both P<0.05). Multivariate analysis showed that serum miR-501 and miR-195 levels were significantly correlated with the sensitivity to concurrent chemoradiotherapy in LACC patients. The area under the ROC curve (AUC) was 0.736 and 0.913, respectively. Conclusions:The higher the serum miR-501 level and the lower the serum miR-195 level before treatment, the higher the probability of resistance to concurrent chemoradiotherapy in LACC patients. Serum miR-501 and miR-195 levels of LACC patients before treatment have certain predictive value for the sensitivity to concurrent chemoradiotherapy.

2.
Article in Chinese | WPRIM | ID: wpr-989861

ABSTRACT

Objective:To investigate the effect and mechanism of miR-195 regulating FOXK1 gene and PI3K/Akt pathway on stomach adenocarcinoma proliferation, invasion and migration ability.Methods:Public database samples were employed to analyze the expression differences and prognostic significance of miR-195 in stomach adenocarcinoma. After overexpression of mir-195-5p in two cell lines, MGC803 and AGS, altered cell proliferation, invasion, and migration abilities were detected by Alamar Blue, Wound healing, and Transwell assays. The potential target genes and binding sites of miR-195 were predicted by the starBase. Western blot was used to detect the expression levels of foxk1 and phosphorylation sites in the PI3K/Akt pathway of target genes after overexpression of mir-195-5p. A Dual-luciferase reporter assay was used to verify the relationship between mir-195-5p and foxk1. Statistical analyses were performed with IBM SPSS 22 software and R 4.0.3.Results:Our results showed a significant over-expression of miR-195 in the tumor tissues, compared with the paired normal tissues ( P<0.001) , which could inhibit the proliferation and invasion of stomach carcinoma cells and significantly correlated with survival ( P=0.011) . Moreover, our study indicated that miR-195 depressed the expression of FOXK1 and significantly reduced the activation of the PI3K/Akt pathway, which had a negative effect on the proliferation and invasion of stomach carcinoma cells. The phosphorylated Akt (s473 site) expression in the PI3K/Akt pathway was significantly decreased after overexpression of miR-195. Conclusion:Overall, our studies clarify the important function of the miR-195 in the diagnosis and therapy of patients with stomach carcinoma and reveal the FOXK1 and PI3K/Akt pathway regulation by the miR-195, which are of important clinical significance in the differential diagnosis.

3.
Article in Chinese | WPRIM | ID: wpr-1014893

ABSTRACT

AIM: To investigate the effect of miR-195-5p on ox-LDL-induced vascular endothelial cell injury and its mechanism. METHODS: The expression of miR-195-5p in HUVECs cells induced by ox-LDL was detected by RT-PCR. Cell proliferation, the expression of oxidative stress associated factors (MDA and LDH) and apoptosis were detected by CCK-8, ELISA and flow cytometry, respectively. The potential targets of miR-195-5p were determined by dual luciferase reporter assay. The expression of target protein in ox-LDL-induced HUVECs cells and the relationship between miR-195-5p and FBXW7 expression were detected by Western blotting. RESULTS: The expression of miR-195-5p in ox-LDL-induced HUVECs cells was significantly higher than that in normal HUVECs cells. Subsequently, miR-195-5p silencing in ox-LDL-induced HUVECs cells effectively promoted the proliferation of HUVECs cells, whereas suppressed the expression of oxidative stress associated factors (MDA and LDH) and apoptosis level. Luciferase reporter assay and Western blot results showed that miR-195-5p could directly target FBXW7 protein gene and negatively regulate its expression. Finally, miR-195-5p modulated the proliferation, oxidative stress and apoptosis of HUVECs cells induced by ox-LDL by regulating the expression of FBXW7. CONCLUSION: miR-195-5p is highly expressed in HUVECs cells induced by ox-LDL. In addition, miR-195-5p can aggravate ox-LDL-induced cytotoxicity, oxidative stress and apoptosis of HUVECs cells by inhibiting the expression of FBXW7.

4.
Article in Chinese | WPRIM | ID: wpr-930265

ABSTRACT

Objective:To investigate the effects of miR-195 on the migration, invasion and epithelial mesenchymal transition (EMT) of human gastric cancer cell line (HGC-27) and its mechanism.Methods:HGC-27 cells were cultured in vitro and they were randomly divided into control group, miR-195 negative control (NC) group and miR-195 mimics (mimics) group; the expressions of miR-195 and SALL4 mRNA were detected by real-time fluorescence quantitative PCR (RT-qPCR) ; The changes in cell morphology of each group were observed under a microscope; the expressions of SALL4, E-cadherin, N-cadherin and Vimentin in HGC-27 cells were detected by Western blot; MTT method was used to detect the change of HGC-27 cell survival rate; the abilities of migration and invasion in HGC-27 cells were detected by scratch test and invasion test (Transwell) ; and the targeting relationship between miR-195 and SALL4 was confirmed by double Luciferase Report.Results:Compared with those in control group and miR-195 NC group, the miR-195 expression level, apoptosis rate and E-cadherin protein expression level of HGC-27 cells in miR-195 mice group were significantly higher ( P<0.05) , the cell survival rate, scratch healing rate, invasion number, SALL4 mRNA and SALL4, N-cadherin, Vimentin protein expression levels were significantly lower ( P<0.05) , the morphology of HGC-27 cells returned to normal, and the epithelial mesenchymal behavior was significantly reduced; in addition, there was a binding site between miR-195 and SALL4 mRNA 3'UTR region, and compared with that in SALL4-3'UTR-WT + miR-195 NC group, the luciferase activity of SALL4-3'UTR-WT + miR-195 MICs group was lower ( P<0.05) . Conclusion:MiR-195 may inhibit the migration, invasion and epithelieal-mesenchymal transition of human gastric cancer cells by targetingly inhibiting the expression of SALL4.

5.
Clinics ; Clinics;76: e2502, 2021. tab, graf
Article in English | LILACS | ID: biblio-1339695

ABSTRACT

OBJECTIVES: Diagnosis and management of essential hypertension (EH) or type 2 diabetes mellitus (T2DM) by combining comprehensive treatment and classificatory diagnosis have been continuously improved. However, understanding the pathogenesis of EH patients with concomitant T2DM and subsequent treatment remain the major challenges owing to the lack of non-invasive biomarkers and information regarding the underlying mechanisms. METHODS: Herein, we collected 200 serum samples from EH and/or T2DM patients and healthy donors (N). Gene-expression profiling was conducted to identify candidate microRNAs with clinical significance. Then, a larger cohort of the aforementioned patients and 50 N were used to identify the correlation between the tumor suppressor miR-195-5p and EH and/or T2DM. The dual-luciferase reporter assay was used to explore the target genes of miR-195-5p. The suppressive effects of miR-195-5p on the 3′-UTR of the dopamine receptor D1 (DRD1) transcript in EH patients with concomitant T2DM were verified as well. RESULTS: Compared with that in other groups, serum miR-195-5p was highly downregulated in EH patients with concomitant T2DM. miR-195-5p overexpression efficiently suppressed DRD1 expression by binding to the two 3′-UTRs. Additionally, two single nucleotide polymorphisms, including 231T-A and 233C-G, in the miR-195-5p binding sites of the DRD1 3′-UTR were further identified. Collectively, we identified the potential clinical significance of DRD1 regulation by miR-195-5p in EH patients with concomitant T2DM. CONCLUSIONS: Our data suggested that miR-195-5p circulating in the peripheral blood served as a novel biomarker and therapeutic target for EH and T2DM, which could eventually help address major challenges during the diagnosis and treatment of EH and T2DM.


Subject(s)
Humans , Receptors, Dopamine D1/genetics , MicroRNAs/genetics , Diabetes Mellitus, Type 2/genetics , Essential Hypertension/genetics , Biomarkers , Polymorphism, Single Nucleotide
6.
Zhonghua nankexue ; Zhonghua nankexue;(12): 875-880, 2020.
Article in Chinese | WPRIM | ID: wpr-880285

ABSTRACT

Objective@#To investigate the regulatory effect of the transcription factor NF-kB1 on the expression of miR-195 in prostate cancer (PCa).@*METHODS@#We analyzed the possibility of NF-kB1 binding to the miR-195 promoter and the expression of NF-kB1 in PCa using the JASPAR and Oncomine databases, respectively, and determined the expressions of NF-kB1 and miR-195 in PCa cells by real-time quantitative PCR after inhibiting the former by interfering RNA targeting NF-kB1. We detected the activity of the luciferase reporter gene after constructing its gene plasmid in the miR-195 promoter region and having it co-transfected with the NF-kB1 plasmid. Then we analyzed the correlation between the expressions of miR-195 and NF-kB1 in the prostate tissue.@*RESULTS@#NF-kB1 was overexpressed in PCa. After inhibition of the expression of NF-kB1, that of miR-195 was increased in PC-3 and DU-145 cell lines, with a negative correlation between the NF-kB1 and miR-195 expressions in the PCa tissue. The results of luciferase reporter gene assay showed direct binding of NF-kB1 to the miR-195 promoter zone.@*CONCLUSIONS@#NF-kB1 regulates the expression of miR-195 in prostate cancer.


Subject(s)
Humans , Male , Cell Line, Tumor , Gene Expression Regulation, Neoplastic , MicroRNAs/genetics , NF-kappa B p50 Subunit/metabolism , Promoter Regions, Genetic , Prostatic Neoplasms/genetics , Transcription Factors/metabolism
7.
J Biosci ; 2019 Dec; 44(6): 1-9
Article | IMSEAR | ID: sea-214213

ABSTRACT

MicroRNAs (miRNAs) have been identified as potential biomarkers for endometrial carcinoma (EC) diagnosis, prognosisand therapy. The purpose of the present study was to investigate the detailed role and molecular mechanism of miR-195 inEC metastasis. qRT-PCR assay was performed to assess the expression of miR-195 and SRY-related high-mobility groupbox 4 (SOX4) mRNA in EC tissues and cells. The levels of N-cadherin, Vimentin, E-cadherin and SOX4 protein weredetermined by western blot. SOX4 protein expression in EC tissues was also determined by Immunohistochemistry (IHC)experiment. Transwell assay was used to analyze cell migration and invasion abilities. Dual-luciferase reporter assay andRNA Immunoprecipitation (RIP) assay were performed to confirm the targeted interaction between miR-195 and SOX4.Our data supported that miR-195 was downregulated and SOX4 was upregulated in EC tissues and cell lines. Upregulationof miR-195 inhibited migration, invasion and epithelial-mesenchymal transition (EMT) of EC cells. Moreover, SOX4 was adirect target of miR-195. MiR-195 overexpression-mediated anti-migration, anti-invasion and anti-EMT effects wereantagonized by SOX4 restoration in EC cells. In conclusion, our study suggested that miR-195 inhibited the migration,invasion and epithelial mesenchymal transition (EMT) of EC cells at least partly by targeting SOX4. Our study provided anovel underlying mechanism for EC metastasis and a promising therapeutic target for EC management.

8.
Article in Chinese | WPRIM | ID: wpr-801069

ABSTRACT

Objective@#To investigate the effect and mechanism of LncRNA ANRIL on the radiosensitivity of HCT116 cells line and nude mouse transplant tumors.@*Methods@#The expression of LncRNA ANRIL in colorectal cancer cells was detected by qPCR. The negative control siRNA, ANRIL siRNA, miR-NC mimic, miR-195 mimic, miR-NC inhibitor and miR-195 inhibitor were transfected into HCT116 cells, and marked as negative control group, silencing ANRIL group, overexpressing miR-NC group, overexpressing miR-195 group, inhibiting miR-NC group and inhibiting miR-195 group, and the HCT116 cells without any treatment were marked as the blank control group. The clone formation assay was used to detect radiosensitivity of colorectal cancer cells, flow cytometry was used to detect apoptosis. The web site, StarBase, was used to predict the downstream miRNAs of ANRIL and dual luciferase reporter gene assay was used to further verify. Subcutaneous tumor transplantation assay was used to detect the effect of ANRIL on the growth of colorectal cancer cells after irradiation.@*Results@#After irradiation with 2, 4, 6 and 8 Gy, the cell survival fraction of silencing ANRIL group was significantly decreased when compared with that of negative control group (P<0.05), and the radiosensitivity ratio was 1.52. The apoptosis rate of the silencing ANRIL+ 4 Gy group was significantly higher than that of the negative control+ 4 Gy group ((27.86±2.78)% vs. (12.06±1.46)%, P<0.05). The results of the experiment on nude mouse transplant tumors showed that the tumor volume in the negative control group was lower than that of the silent ANRIL group on days 13, 16, 19, 22 and 25 ((234±66) mm3, (273±63) mm3, (296±72) mm3, (321±85) mm3 and (403±94) mm3 vs. (357±79) mm3, (485±124) mm3, (617±143) mm3, (764±174) mm3 and (985±221) mm3P<0.05). MiR-195 is a target gene of ANRIL, and inhibition of miR-195 can reverse the inhibitory effect of silencing ANRIL on radiosensitivity, apoptosis and xenografts of HCT116 cells.@*Conclusions@#LncRNA ANRIL regulates the radiosensitivity of colorectal cancer cells by miR-195, which may provide a new sensitizing target for clinical colorectal cancer radiotherapy.

9.
Article in Chinese | WPRIM | ID: wpr-798333

ABSTRACT

@# Objective: To explore the effect of miR-195/TLR4 axis on the proliferation, invasion and migration of liver cancer cells via regulating NF-κB pathway. Methods: Twenty-five pairs of liver cancer tissues and corresponding adjacent tissues surgically resected at the Second Affiliated Hospital of Kunming Medical University from March 2016 to January 2017 were collected for this study. Liver cancer HepG2 cells were cultured and then randomly divided into four groups: control group (NC), miR-195 mimic group (miR-195), TLR4 knockdown group (si-TLR4), and miR-195 inhibitor combined with TRL4 knockdown group (si-TLR4+miR-195 inhibitor). qRTPCR was used to detect the expression of miR-195 in liver cancer tissues and cell lines. CCK-8 assay was used to evaluate the cell viability of each group. Transwell and Wound healing assay were applied to detect the invasion and migration ability of HepG2 cells, respectively. Dual-luciferase reporter gene assay was used to verify the targeted regulation of TLR4 by miR-195. WB was applied to analyze the protein expressions of TLR4 and NF-κB p65. Results: miR-195 was down-regulated in the liver cancer tissues compared with adjacent tissues (P<0.01). Compared with human hepatic epithelial cells (THLE-3), the expression of miR-193 in liver cancer cell lines (HepG2 and Huh-7) was down-regulated (P<0.01), and the expression level in HepG2 cells was the lowest. The proliferation, invasion and migration of HepG2 cells was significantly suppressed after over-expression of miR-195 (all P<0.01). Moreover, over-expression of miR-195 significantly down-regulated TLR4 protein expression (P<0.05), and TLR4 was negatively correlated with miR-195 (R2= 0.602, P<0.0001). Furthermore, miR-195 over-expression inhibited proliferation, invasion and migration of HepG2 cells by targeting TLR4 expression and blocking NF-κB pathway (P<0.05 or P<0.01). Conclusion: miR-195 over-expression can inhibit the proliferation, invasion and migration of HepG2 cells. The mechanism may be related with targeting TLR4 and blocking the NF-κB pathway to affect cell biological behaviors.

10.
Article in Chinese | WPRIM | ID: wpr-812027

ABSTRACT

@# Objective: To explore the molecular mechanism of miR-195-5p targeting FGF2 to inhibit the proliferation, apoptosis, invasion and migration of endometrial cancer HEC-1B cells. Methods: After culture and transfection, HEC-1B cells were divided into 4 groups: HEC-1B group, miR-195-5p mimic group, pLV-FGF2 group and miR-195-5p+FGF2 group. The expressions of miR-195-5p and mRNA levels of FGF2 were detected by qRT-PCR. The targeted relationship of miR-195-5p and FGF2 was verified by luciferase assay. The protein expression of FGF2 was examined by Western blotting; Proliferation of HEC-1B cells was measured by CCK-8; Apoptosis was tested by flow cytometry; HEC-1B cell invasion was detected by transwell, and migration was measured by scratch assay. Results: Compared with HEC-1B group, the expression of miR-195-5p in miR-195-5p mimic group was elevated while FGF2 mRNA level was declined (all P<0.01). Luciferase assay indicated that FGF2 was a target of miR-195-5p. Compared with HEC-1B group, the protein level of FGF2 in miR-195-5p mimic group was decreased, and the protein levels of FGF2 in pLV-FGF2 group were enhanced (P<0.01). The protein levels of FGF2 in miR-195-5p+FGF2 group were lower than that in pLV-FGF2 group (all P<0.01). The proliferation in miR-195-5p mimic group was lower than HEC-1B group (P<0.01), while the proliferation in pLV-FGF2 group was higher than that in HEC-1B group (all P<0.01). Compared with HEC-1B group, apoptosis in miR-195-5p mimic group was increased, and apoptosis in pLV-FGF2 group was decreased (P<0.01); moreover, apoptosis in miR-195-5p+FGF2 group was higher than that in pLV-FGF2 group (P<0.01). Compared with HEC-1B group, the number of invasive cells per field and the rate of wound healing in miR195-5p mimic group were decreased, while those in pLV-FGF2 group was enhanced (P<0.01); moreover, the number of invasive cells per field and the rate of wound healing in miR-195-5p+FGF2 group was lower than those in pLV-FGF2 group (all P<0.01). Conclusion: miR-195-5p inhibits proliferation, invasion and migration and promotes apoptosis of endometrial cancer HEC-1B cells by targeting FGF2, and could be used as a treatment target of endometrial cancer.

11.
Article in Chinese | WPRIM | ID: wpr-843667

ABSTRACT

Schizophrenia is a serious mental health illness with unknown etiology, high prevalence, high morbidity and heavy social burden. Some RNAs with regulatory functions may be involved in pathogenesis of many complex diseases, including schizophrenia. miRNAs are non-coding RNAs that negatively regulate gene expression. Differential miRNA expression has been found in the brain and peripheral blood of patients with schizophrenia. miR-195 is an important member of the miR-15/16/195/497 family. Previous studies have implicated that miR-195 may be involved in the pathogenesis of schizophrenia due to its differential expression. This review aimed to clarify the function of miR-195 and summarized the recent researches of miR-195 in schizophrenia.

12.
The Journal of Practical Medicine ; (24): 3698-3701, 2017.
Article in Chinese | WPRIM | ID: wpr-697507

ABSTRACT

Objective To investigate the effects of inhibition S6K1 by miR-195 on proliferation,apoptosis,invasion and metastasis of prostate cancer cells.Method The prostate cancer cell line DU145 and LNCaP were used to establish cell lines with overexpression or suppression of miR-195.S6K1 mRNA expression was determined by the reverse transcription PCR (RT-PCR) assay.Cell proliferation was detected by MTT assay,cell apoptosis was detected by flow cytometry,cell invasion and metastasis was detected by transwell invasion test and cell scratch test,respectively.Target gene of miR-195 was identified by using dual luciferase reporter assay.Results Cell proliferation rate in miR-195 overexpression group was significantly lower than that in the control group.Cell apoptosis in miR-195 overexpression group was higher than that in the control group.The invasive cell numbers and scratch distance in miR-195 overexpression group were less than those in the control group,but the invasive cell numbers and scratch distance were increased in miR-195 knockdown group.S6K1 expression could be inhibited by miR-195,and S6K1 was identified as a target gene of miR-195.Conclusion miR-195 regulates proliferation,apoptosis,invasion and metastasis of prostate cancer cell by targeting S6K1.

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