The lncRNA-MALAT1/miR-330-3p Axis Promotes Growth and Metastasis of Gastric Cancer Through Regulation of Vascular Endothelial Growth Factor A: A Genetic and Cell Evaluation Study

SUMMARY: Overexpression of metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) in various tumor tissues and cell lines was found to promote tumor cell proliferation, migration, and invasion. However, the role of MALAT1 in gastric cancer (GC) is still unclear. We aimed to investigate the correlation between long-chain non-coding RNAs (lncRNAs), MALAT1, MicroRNAs (miRNA) and vascular endothelial growth factor A (VEGFA) in gastric cancer and to disclose underlying mechanism. The correlation between MALAT1 levels and clinical features was analyzed by bioinformatics data and human samples. The expression of MALAT1 was down regulated in AGS cells to detect the cell proliferation, migration, and invasion characteristics, as well as the effects on signal pathways. Furthermore, we validated the role of MALAT1/miR-330-3p axis in GC by dual luciferase reporter gene assays. Expression of MALAT1 was higher in cancer tissues than in para-cancerous tissues. The high MALAT1 level predicted malignancy and worse prognosis. Down-regulation of MALAT1 expression in AGS cells inhibited cell proliferation, migration, and invasion by targeting VEGFA. By dual luciferase reporter gene assay and miR-330-3p inhibitor treatment, we demonstrate that MALAT1 sponged miR-330-3p in GC, leading to VEGFA upregulation and activation of the mTOR signaling pathway. The MALAT1/miR-330-3p axis regulates VEGFA through the mTOR signaling pathway and promotes the growth and metastasis of gastric cancer.

Se descubrió que la sobreexpresión del transcrito 1 de adenocarcinoma de pulmón asociado a metástasis (MALAT1) en varios tejidos tumorales y líneas celulares promueve la proliferación, migración e invasión de células tumorales. Sin embargo, el papel de MALAT1 en el cáncer gástrico (CG) aún no está claro. Nuestro objetivo fue investigar la correlación entre los ARN no codificantes de cadena larga (lncRNA), MALAT1, los microARN (miARN) y el factor de crecimiento endotelial vascular A (VEGFA) en el cáncer gástrico y revelar el mecanismo subyacente. La correlación entre los niveles de MALAT1 y las características clínicas se analizó mediante datos bioinformáticos y muestras humanas. La expresión de MALAT1 se reguló negativamente en las células AGS para detectar las características de proliferación, migración e invasión celular, así como los efectos sobre las vías de señales. Además, validamos el papel del eje MALAT1/miR- 330-3p en GC mediante ensayos de genes indicadores de luciferasa dual. La expresión de MALAT1 fue mayor en tejidos cancerosos que en tejidos paracancerosos. El alto nivel de MALAT1 predijo malignidad y peor pronóstico. La regulación negativa de la expresión de MALAT1 en células AGS inhibió la proliferación, migración e invasión celular al apuntar a VEGFA. Mediante un ensayo de gen indicador de luciferasa dual y un tratamiento con inhibidor de miR-330-3p, demostramos que MALAT1 esponjaba miR-330-3p en GC, lo que lleva a la regulación positiva de VEGFA y la activación de la vía de señalización mTOR. El eje MALAT1/miR-330-3p regula VEGFA a través de la vía de señalización mTOR y promueve el crecimiento y la metástasis del cáncer gástrico.

Expression relationship and significance of NEAT1 and miR-27a-3p in serum and cerebrospinal fluid of patients with Alzheimer disease / 北京大学学报(医学版)

Objective:To explore the expression relationship and significance of long chain non-coding RNA nuclear-enriched abundant transcript 1(LncRNA NEAT1)and miR-27a-3p in serum and cerebro-spinal fluid of patients with Alzheimer disease(AD).Methods:Sixty-six AD patients received by the department of neurology of our hospital from October 2019 to September 2021 were gathered,according to the clinical dementia rating scale score,they were grouped into mild group(≤ 1 point,n=41)and moderate-to-severe group(＞1 point,n=25).Another 66 cases of serum and cerebrospinal fluid sam-ples from outpatient physical examination personnel were regarded as the control group.The general infor-mation on all subjects was recorded and cognition was assessed;real-time quantitative PCR was performed to measure the expression levels of miR-27a-3p and NEAT1 in serum and cerebrospinal fluid;enzyme-linked immunosorbent assay was performed to measure the protein levels of β-amyloid precursor protein cleaving enzyme 1(BACE1),β-amyloid(Aβ)40 and Aβ42 in cerebrospinal fluid;Spearman's method was performed to analyze the correlation of serum miR-27a-3p and NEAT1 levels with mini-mental state examination(MMSE)and montreal cognitive assessment(MoCA)scores;Pearson method was per-formed to analyze the correlation between serum miR-27a-3p and NEAT1 levels and Aβ deposition standard uptake value ratio(SUVR)and cerebrospinal fluid miR-27a-3p,NEAT1,BACE1,Aβ42 and Aβ40 levels.Results:The MMSE score[21(17,25),9(7,11)vs.27(21,34)],MoCA score[17(12,21),10(7,13)vs.27(21,31)],serum miR-27a-3p level(0.55±0.13,0.46±0.06 vs.0.97± 0.22),cerebrospinal fluid miR-27a-3p(0.48±0.10,0.35±0.10 vs.1.03±0.31),Aβ42 levels[(303.55±36.77)ng/L,(231.45±34.14)ng/L vs.(499.99±53.63)ng/L]and Aβ42/Aβ40 ra-tio(0.030±0.008,0.022±0.007 vs.0.048±0.010)of AD patients in mild group and moderate-to-severe group were all lower than those in the control group,and the moderate-to-severe group were lower than the mild group(all P＜0.05);the serum NEAT1 level(2.31±0.64,3.13±0.76 vs.1.05± 0.20),SUVR(1.50±0.29,1.76±0.52 vs.0.74±0.15),and cerebrospinal fluid NEAT1(3.51± 1.24,4.30±1.65 vs.1.01±0.23)and B ACE 1 levels[(55.78±5.98)μg/L,(72.32±16.08)μg/L vs.(21.39±3.73)μg/L]were higher than those in the control group,and the moderate-to-se-vere group were higher than the mild group(all P＜0.05).Serum NEAT1 level in AD patients was posi-tively correlated with SUVR,cerebrospinal fluid NEAT1 and BACE1(r=0.350,0.606,0.341,P＜0.05),and negatively correlated with MMSE score and MoCA score(r=-0.473,-0.482,all P＜0.05);serum miR-27a-3p level was positively correlated with cerebrospinal fluid miR-27a-3p level,MMSE score and MoCA score(r=0.695,0.424,0.412,all P＜0.05),and negatively correlated with SUVR and cerebrospinal fluid BACE1 level(r=-0.521,-0.447,all P＜0.05).Conclusion:The expression trends of NEAT1 and miR-27a-3p in the serum and cerebrospinal fluid of AD patients are con-sistent,the level of NEAT1 is increased,and the level of miR-27a-3p is decreased.The levels of the two are negatively correlated,which is related to the degree of Aβ deposition in the brain of AD patients and is involved in the progression of AD.

Expression of serum lncRNA CCDC18-AS1 and miR-501 in patients with HPV-infected cervical cancer and its correlation with prognosis / 国际检验医学杂志

Objective To explore the expression of serum long noncoding RNA(lncRNA)CCDC18-AS1 and microRNA-501(miR-501)in patients with human papillomavirus(HPV)-infected cervical cancer,and its correlation with prognosis.Methods A total of 78 patients with HPV-infected cervical cancer who underwent treatment in the hospital from May 2019 to May 2020 were enrolled as study group,meantime,80 patients with simple HPV infection and 81 patients with simple non-HPV infected cervical cancer were set as control group 1 and control group 2.Serum lncRNA CCDC18-AS1 and miR-501 expression levels were detected by re-al-time quantitative PCR.Then the clinical features,postoperative overall survival and progression-free surviv-al were analyzed in the study group.Results The relative expression levels of serum lncRNA CCDC18-AS1 and miR-501 in the study group were higher than those in control group 1 and control group 2(P<0.05),and the relative expression levels of serum lncRNA CCDC18-AS1 and miR-501 in control group 2 were higher than those in control group 1(P<0.05).Pearson correlation analysis showed that serum lncRNA CCDC18-AS1 and miR-501 were positively correlated in the study group(r=0.421,P<0.001).Serum lncRNA CCDC18-AS1 and miR-501 in study group were not related to age,but were related to clinical stage,differentiation de-gree,lymph node metastasis,and squamous epithelial cell antigen(P<0.05).Kaplan Meier analysis showed that the postoperative overall survival and progression-free survival of high lncRNA CCDC18-AS1 and miR-501 expression groups were shorter than those of low expression groups(P<0.05).Conclusion HPV-infec-ted cervical cancer patients have up-regulated expression of serum lncRNA CCDC18-AS1 and miR-501,and the abnormal changes are closely related to patients'clinical characteristics and postoperative survival,so the two indicators could be used as auxiliary indicators for the disease assessment and prognosis evaluation.

Study on the correlation between the active components of Salviea Miltiorrhizae Radix et Rhizoma screened by high-throughput sequencing and the regulation of lncRNA-mRNA in human lung adenocarcinoma A549 cells / 国际中医中药杂志

Objective:Study on the correlation between the active components of Salviea Miltiorrhizae Radix et Rhizoma screened by high-throughput sequencing and the regulation of lncRNA-mRNA in human lung adenocarcinoma A549 cells.Methods:A549 cells were cultured, and the IC 50 dose of cryptotanshinone and tanshinone ⅡA was confirmed according to the cell proliferation experiment. A549 cells were randomly divided into blank control group, cryptotanshinone group, and tanshinone IIA group using a random number table method. After 24 hours of intervention, the cell cycle was detected by flow cytometry. High-throughput sequencing technique was used to detect the expressions of lncRNA and mRNA in A549 cells in intervention group and non-intervention group. By analyzing the expression profiles of differential genes related to cryptotanshinone and tanshinone ⅡA, the obtained differential genes were analyzed by GO and KEGG. Results:The cell cycle results showed that the proportion of G0/G1 phase cells in cryptotanshinone and Tanshinone ⅡA was increased ( P<0.01), the proportion of S phase cells was decreased ( P<0.01), and the proportion of G2/M phase cells in cryptotanshinone was decreased ( P<0.01). The results of high-throughput screening showed that cryptotanshinone could up-regulate 4 698 lncRNA, down-regulate 1 557 lncRNA, up-regulate 4 810 mRNA and down-regulate 5 644 mRNA. Tanshinone ⅡA could up-regulate 1 348 lncRNA, down-regulate 1 299 lncRNA, up-regulate 4646 mRNA and down-regulate 4 741 mRNA. The function and pathway enrichment analysis of differential lncRNA and mRNA showed that the differentially expressed genes of cryptotanshinone and tanshinone ⅡA were mainly related to cell cycle, autophagy, AMPK signaling pathway, FoxO signaling pathway and EGFR signaling pathway. GAS5 may be one of the targets for the inhibitory effects of cryptotanshinone and tanshinone ⅡA. Conclusion:Cryptotanshinone and tanshinone ⅡA have certain inhibitory effects on A549 cells, and there are differentially expressed genes of lncRNA-mRNA, which are closely related to cell cycle and signal pathway.

lncRNA VIM-AS5 expression and its effect on proliferation and migration of human breast cancer cell lines / 基础医学与临床

Objective To explore the clinical significance of long non-coding RNA(lncRNA)VIM-AS5 expres-sion in human breast cancer tissues and its regulatory mechanism involved in cancer cell proliferation and mi-gration.Methods The Lnc2Cancer 3.0 database was used to analyze the expression of VIM-AS5 in breast cancer tissues and its correlation with the clinical stage and survival time of breast cancer patients.RT-qPCR was used to detect the expression of VIM-AS5 in breast cancer cell lines BT-549,MDA-MB-435,MDA-MB-231 and CAL-51.Plasmid with VIM-AS5 overexpression and negative control were all transfected into CAL-51 cells through liposome recorded as VIM-AS5 group and NC group,respectively.The proliferation and migration of CAL-51 cells were detected by colony formation assay and scratch healing method,respectively.Dual-lucif-erase reporter gene experiment verified the targeting relationship between VIM-AS5 and miR-500a.RT-qPCR was used to detect the expression of miR-500a in CAL-51 cells.Western blot was used to detect the expression of JAK/STAT3 pathway in CAL-51 cells.Results The expression of VIM-AS5 in breast cancer tissues was significantly lower than that in adjacent tissues(P＜0.01).VIM-AS5 expression was negatively correlated with the clinical stage of breast cancer patients(P＜0.01).The survival time of breast cancer patients with low VIM-AS5 expression was significantly shorter than that of breast cancer patients with high VIM-AS5 ex-pression(P＜0.01).Compared with mammary epithelial cell line MCF-10 A cells,VIM-AS5 expression was significantly reduced in breast cancer cells(P＜0.01).The counting number of colony formed in the VIM-AS5 group was significantly lower than that in the NC group(P＜0.01).The cell migration rate in the VIM-AS5 group was significantly lower than that in the NC group(P＜0.01).Dual-luciferase reporter gene experiment confirmed that miR-500a was the target gene of VIM-AS5(P＜0.01).VIM-AS5 can negatively regulate the expression of miR-500a(P＜0.01).Compared with the NC group,the expression of JAK/STAT3 pathway proteins JAK,p-STAT3,c-Myc,Bcl-2,and CDK3 in CAL-51 cells of the VIM-AS5 group were significantly decreased.Conclusions VIM-AS5 is low-expressed in breast cancer cells,and up-regulation of VIM-AS5 may inhibit the proliferation and migration of breast cancer cells CAL-51 by targeting at miR-500a/JAK/STAT3 pathway.

Effect of LncRNA MEG3 on blood-brain barrier in neonatal mice with hypoxic-ischemic brain injury by targeting miR-17-5p / 临床神经病学杂志

Objective To analyze the effect of LncRNA MEG3 on the blood-brain barrier in neonatal mice with hypoxic-ischemic brain injury(HIBD)by regulating miR-17-5p.Methods A total of 90 C57BL mice were randomly grouped into control group,model group,si-NC group,si-LncRNA MEG3 group,si-LncRNA MEG3+anti-miR-NC group and si-LncRNA MEG3+anti-miR-17-5p group,with 15 mice in each group.Except for the control group,the rest of the mice were constructed with HIBD model,and the Longa score was used to evaluate the nerve damage of mice.The ultrastructure of vascular endothelial cells around brain tissue was observed by electron microscope.The permeability of blood-brain barrier in mice was measured by EB method.The levels of LncRNA MEG3 and miR-17-5p in the brain tissue of mice in each group were determined by qRT-PCR.The levels of ZO-1 and Occludin protein in the brain tissue of mice were determined by Western blotting.Results Compared with control group,the vascular endothelium of mice in model group was unevenly thin and thick,and showed edema in many places.Compared with model group,the vascular endothelial cells in the si-LncRNA MEG3 group were gradually tightly connected,the thickness was more uniform,and the edema was reduced.Compared with the si-LncRNA MEG3 group,vascular endothelial cell damage was intensified in the si-LncRNA MEG3+anti-miR-17-5p group.Compared with those in the control group,the neurological deficit score,brain water content,EB content,and LncRNA MEG 3 level in the model group were significantly increased,miR-1 7-5 p level,ZO-1 and Occludin expression were significantly decreased(all P＜0.05).Compared with those in model group,there were no significant differences in neural function deficit score,brain water content,EB content,lncRNA MEG3 level,miR-17-5p level,ZO-1 and Occludin expression in si-NC group(all P＞0.05);the neurological deficit score,brain water content,EB content and LncRNA MEG3 level in si-LncRNA MEG3 group were significantly decreased,miR-17-5p level,ZO-1,and Occludin expression were significantly increased(all P＜0.05).Compared with those in si-LncRNA MEG3 group,there were no significant differences in neural function deficit score,brain water content,EB content,LncRNA MEG3 level,miR-17-5p level,ZO-1 and Occludin expression in si-LncRNA MEG3+anti-miR-NC group(all P＞0.05);the neurological deficit score,brain water content and EB content in si-LncRNA MEG3+anti-miR-17-5p group were significantly increased,miR-17-5p level,ZO-1 and Occludin expression were significantly decreased(all P＜0.05).Conclusion LncRNA MEG3 silencing may reduce the permeability of the blood-brain barrier in neonatal mice with HIBD by up-regulating miR-17-5p,maintaining the protective effect of the blood-brain barrier on the brain,and protecting brain tissue.

Advances in epigenetics of nonalcoholic fatty liver desease / 复旦学报（医学版）

Nonalcoholic fatty liver disease(NAFLD)is a metabolic liver disease that ranges from relatively benign hepatic steatosis to nonalcoholic steatohepatitis(NASH).NASH is characterized by persistent liver damage,inflammation,and fibrosis which significantly increases the risk of end-stage liver diseases,such as liver cirrhosis and hepatocellular carcinoma.The pathogenesis of NAFLD/NASH is not yet fully understood,but its recent epigenetic advances have provided new insights into the mechanisms of this disease.This review summarized recent progress in this area which has laid a solid foundation for elucidating the pathogenesis of NAFLD and provides potential targets for early detection,diagnosis,and treatment of this disease.

Study on the Relationship between Serum LncRNA SNHG11 and HIF-1α Level Expression and Vasculogenic Mimicry of Cancer Tissues in Ovarian Cancer / 现代检验医学杂志

Objective To investigate the relationship between the expression of long non-coding RNA(LncRNA)small nucleolar RNA host gene 11(SNHG11)and hypoxia inducible factor(HIF)-1α and angiogenesis mimicry(VM)in ovarian cancer.Methods A total of 116 ovarian cancer patients admitted to Tangshan Maternal and Child Health Care Hospital from October 2019 to January 2023 were regarded as the research subjects.Based on whether VM had formed,ovarian cancer patients were grouped into VM group(n=51)and non VM group(n=65).Another 50 partients who underwent health examinations during the same period were regarded as the control group.Real-time fluorescence quantitative PCR(qPCR)was applied to detect the expression levels of LncRNA SNHG11 and HIF-1α in serum of ovarian cancer patients and control groups.Spearman correlation was applied to detect the relationship between LncRNA SNHG11,HIF-1α,and VM formation.The diagnostic value of LncRNA SNHG11,HIF-1α,and their combined detection in the formation of VM in ovarian cancer patients was analyzed using the receiver operating characteristic(ROC)curve.Results Compared with the control group,the levels of serum LncRNA SNHG11(3.01±0.88,2.21±0.68 vs 1.12±0.35)and HIF-1α(2.16±0.67,1.60±0.44 vs 1.01±0.31)in ovarian cancer patients with VM group and non VM group were increased(t=12.136,9.006;19.890,16.591,all P＞0.05),the levels of serum LncRNA SNHG11 and HIF-1αin the VM group were obviously higher than those in the non VM group(t=8.957,8.595),and the differences were statistically significant(all P＜0.05).The expression of LncRNA SNHG11,HIF-1α,and the formation of VM were not related to age and tissue type(t=1.036,0.976,0.218;1.254,1.390,0.368,all P＞0.05),but were related to tumor size,FIGO staging,lymph node metastasis,and pathological grading(t=5.351,5.186,13.264;5.465,5.227,10.898;6.063,6.016,5.374;4.030,5.871,5.509,all P＜0.05).Spearman correlation analysis showed that there were obvious positive correlations between LncRNA SNHG11,HIF-1α,and VM generation(r=0.560,0.494,all P<0.05).ROC curve results showed that the areas under the curve(AUCs)of serum LncRNA SNHG11 and HIF-1α for diagnosing VM formation in ovarian cancer patients were 0.860 and 0.824,respectively,with sensitivity of 80.4%and 75.6%,specificity of 58.9%and 51.9%,respectively.The AUC of VM formation in ovarian cancer patients diagnosed by the combination of the two was 0.941,with sensitivity and specificity were 92.2%and 79.9%,respectively.Conclusion The abnormal expressions of LncRNA SNHG11 and HIF-1α were closely related to the formation of VM in ovarian cancer patients,and both may serve as potential biological indicators for judging VM.

Expression of lncRNA SNHG8 in placenta accreta and its effect on trophoblast invasion and migration / 实用医学杂志

Objective To investigate the expression of lncRNA SNHG8 in placenta accrete(PA)and its effect on trophoblast invasion and migration.Methods qRT-PCR was used to detect the expression of lncRNA SNHG8 in placenta tissue of 30 cases in PA group and 30 cases in control group,and the correlation between lncRNA SNHG8 expression and prenatal ultrasound score of 30 cases in PA group was analyzed.Transwell and scratch assay were used to detect the effect of lncRNA SNHG8 interference on the invasion and migration of human chorionic trophoblast cells(HTR8/SVneo cells),and western blot was used to detect the expression of MMP-2 and MMP-9.The downstream targets of lncRNA SNHG8 were predicted by StarBase software,and the expression of lncRNA SNHG8 was detected in placental tissues of the two groups.Dual luciferase reporter assay was used to detect the targeting relationship between lncRNA SNHG8 and miR-542-3p.Results Compared with that of the control group,the expression of lncRNA SNHG8 was up-regulated in the placenta tissue of the PA group(P<0.05),and it was positively correlated with prenatal ultrasound score.Interference with lncRNA SNHG8 inhibited the invasion and migration of trophoblast cells(P<0.05);the protein expression of MMP-9 and MMP-2 also decreased signifi-cantly(P<0.05).Biological prediction indicates that miR-542-3p had a binding site with lncRNA SNHG8,and miR-542-3p expression was down-regulated in PA placental tissue(P<0.05).Dual luciferase reporter assay confirmed that lncRNA SNHG8 could target miR-542-3p.Compared with si-SNHG8+inhibitor-NC,co-transfection of si-SNHG8 and miR-542-3p inhibitor enhanced the invasion and migration ability of trophoblast cells(P<0.05).Conclusion lncRNA SNHG8 is highly expressed in PA and is related to the severity of PA.LncRNA SNHG8 promotes the invasion and migration of trophoblast by regulating the level of miR-542-3p.The study suggests that lncRNA SNHG8 plays an important role in the invasion and migration of PA trophoblast cells,which is expected to be a clinical diagnostic biomarker and therapeutic target.

Extracellular vesicles carrying non-coding RNA regulate the activation of osteoclasts / 中国组织工程研究

BACKGROUND:It has been demonstrated that osteoclast activation plays an important role in skeletal system-related diseases.The mechanism of regulation of osteoclast activation by extracellular vesicles carrying non-coding RNA has not been fully elucidated. OBJECTIVE:To review and summarize relevant literature in and outside China,and to review the regulation of osteoclast activation by different non-coding RNAs in extracellular vesicles in different diseases,so as to provide a certain direction for subsequent research. METHODS:"Non-coding RNA,miRNA,lncRNA,circRNA,snoRNA,osteoclasts,extracellular vesicles,exosome,microparticle,apoptotic bodies"were used as search terms to search the databases of CNKI,WanFang,and VIP."Extracellular vesicles,exosome,microparticle,apoptotic bodies,non-coding RNA,miRNA,lncRNA,circRNA,snoRNA,osteoclast"were used as search terms to search PubMed.Finally,71 articles were included. RESULTS AND CONCLUSION:(1)The activation of osteoclasts is affected by many factors,among which the specific mechanism of non-coding RNA regulating osteoclast activation is not clear.(2)Extracellular vesicles can be secreted by osteoblasts,bone marrow mesenchymal stem cells,tumor cells and other cells.As a carrier of intercellular communication,extracellular vesicles can carry non-coding RNA to regulate osteoclast activation.(3)In the current studies on the regulation of osteoclast activation by extracellular vesicles carrying non-coding RNA,most of the diseases are osteoporosis,followed by tumor bone metastasis,and most types of non-coding RNA are miRNA.(4)There are relatively few studies on the regulation of extracellular vesicles carrying lncRNA and circRNA and snoRNA on osteoclast activation,and the regulatory mechanism is mainly ceRNA mechanism.(5)In conclusion,an in-depth study of the regulatory mechanism of extracellular vesicles carrying non-coding RNA on osteoclast activation is helpful to find key targets for the treatment of skeletal system-related diseases.

Role and mechanism of noncoding RNA in diabetic peripheral neuropathy / 中国组织工程研究

BACKGROUND:Persistent hyperglycemia has been identified as promoting neurovascular dysfunction,leading to irreversible endothelial dysfunction,increased neuronal apoptosis,oxidative stress and inflammation.These changes in combination or alone lead to microvascular and macrovascular lesions as well as progressive neuropathy.Noncoding RNAs may provide a new strategy for understanding the etiology,pathogenesis and treatment of the disease. OBJECTIVE:To review the role and mechanism of noncoding RNAs in the occurrence and development of diabetic peripheral neuropathy by reviewing relevant literature at home and abroad,in order to provide new ideas and approaches for noncoding RNAs in the prevention,diagnosis and treatment of diabetes neuropathy. METHODS:CNKI and PubMed were retrieved for relevant literature published from database inception to 2022.The key words were"noncoding RNA;lncRNA;miRNA;diabetes peripheral neuropathy;expression profile"in Chinese and English,respectively.The retrieved documents were summarized and analyzed,and 61 articles were finally selected for further review. RESULTS AND CONCLUSION:(1)Noncoding RNA plays a key role in the pathophysiological process of diabetic peripheral neuropathy.Among the most widely studied regulatory noncoding RNA species,there are long noncoding RNAs,circular RNAs and microRNAs.(2)Through the regulation of noncoding RNAs,the activation or inhibition of related cell pathways,inflammatory genes and downstream-related cytokines will inhibit cell apoptosis,improve inflammation,and thus change the expression of target genes to participate in the process of diabetic neuralgia.(3)Although many microRNAs and long noncoding RNAs have been found to participate in diabetic peripheral neuropathy,the mechanisms of many noncoding RNAs are unclear,and the same noncoding RNAs may play different roles in different modes.Therefore,it is necessary to further study their action modes in disease etiology and pathology,thereby clarifying their role in the pathogenesis of diabetic peripheral neuropathy.However,the criteria for evaluating noncoding RNA activity have not yet been established,and further research is needed on which specific noncoding RNAs play a dominant regulatory role.(4)MicroRNAs,long noncoding RNAs and their target genes can regulate progressive neuropathy,which are expected to become new targets for the clinical prevention and treatment of diabetic peripheral neuropathy and new biomarkers for the development and prognosis of diabetic peripheral neuropathy.

Mechanism of NONHSAT248596.1 endogenous competition with miR-146a-5p regulating osteoarthritis cartilage degeneration / 中国组织工程研究

BACKGROUND:Currently,there have been studies on the regulatory mechanism of lncRNA\miRNA\mRNA co-expression network on the occurrence and development of osteoarthritis.Our research group has screened qualified NONHSAT248596.1 and miR-146a-5p through the database in the previous stage,but the corresponding in vivo experiments to verify the above regulatory mechanisms are still lacking. OBJECTIVE:To explore the role of NONHSAT248596.1 in regulating competitive endogenous RNA of miR-146a-5p in cartilage degeneration mediated by stromal cell derived factor type 1/chemokine receptor 4 axis in vivo. METHODS:The models of osteoarthritis were established in 36 New Zealand rabbits by injecting stromal cell derived factor 1 solution into the knee joint of the right hind limb.According to the random number table method,they were divided into four groups.lncRNA group,miRNA group,ceRNA group and control group were injected with lentivirus vector overexpressing NONHSAT248596.1,lentivirus vector overexpressing miR-146a-5p,lentivirus vector overexpressing miR-146a-5p+NONHSAT248596.1 and empty lentivirus vector into the molded knee joint,respectively.At 4,8 and 12 weeks of modeling,cartilage tissues and subchondral bone tissues of the knee joint were taken for relevant detection. RESULTS AND CONCLUSION:Hematoxylin-eosin staining and safranin fast green staining showed different degrees of degeneration in the four groups.At 4 weeks,the cartilage tissue of the lncRNA group showed swelling of chondrocytes,loss of cell polarity,destruction of extracellular matrix,surface erosion,fracture formation and partial or full layer loss of cartilage tissue.The degree of cartilage injury was gradually aggravated with time.The progression of articular cartilage inflammation in the miRNA group was the slowest among the four groups.qRT-PCR showed that at the same time point,mRNA expression levels of NONHSAT248596.1,chemokine receptor 4,matrix metalloproteinase 3,matrix metalloproteinase 9 and matrix metalloproteinase 13 in cartilage tissue of the lncRNA group were higher than those of the other three groups(P＜0.05).The mRNA expression levels of miR-146a-5p,aggrecan and type Ⅱ collagen were lower than those of the other three groups(P＜0.05).The mRNA expression levels of NONHSAT248596.1,chemokine receptor 4,matrix metalloproteinase 3,matrix metalloproteinase 9 and matrix metalloproteinase 13 in the miRNA group were lower than those in the ceRNA group and control group at 8 and 12 weeks after the model construction(P＜0.05).The mRNA expressions of miR-146a-5p,aggrecan and type Ⅱ collagen were higher than those of the ceRNA group and control group(P＜0.05).Western blot assay showed that at the same time point,the expression levels of aggrecan and type Ⅱ collagen in cartilage tissue of the lncRNA group were always lower than those of the other three groups(P＜0.05).The expression levels of aggrecan and type Ⅱ collagen in cartilage tissue of the miRNA group at 8 and 12 weeks after modeling were higher than those of the ceRNA group and control group(P＜0.05).The results showed that miR-146a-5p,as the target of NONHSAT248596.1,could be inhibited by the effect of its ceRNA.After acting on miR-146a-5p,NONHSAT248596.1 regulates the stromal cell derived factor type 1/chemokine receptor 4 axis to affect the expression of matrix metalloprotein,type Ⅱ collagen,and aggrecan in osteoarthritis chondrocytes,resulting in the degradation of extracellular matrix and the loss of proteoglycan.

Mechanisms of long non-coding RNA in osteoarthritis and traditional Chinese medicine intervention / 中国组织工程研究

BACKGROUND:The etiology of osteoarthritis is varied and its pathogenesis is still unclear.As bioinformatics has been deepening in recent years,increasing studies have found that the aberrant expression of long non-coding RNAs(lncRNAs)and microRNAs(miRNAs)in joint tissues may mediate the downstream signaling pathways involved in the development of osteoarthritis. OBJECTIVE:To review the mechanism of lncRNA in the development of osteoarthritis and the therapeutic effects of monomers and active compounds derived from traditional Chinese medicine that modulate lncRNA and downstream signaling pathways in osteoarthritis. METHODS:We searched CNKI,WanFang,VIP,and PubMed using the search terms of"long non-coding RNA,knee osteoarthritis,miRNA,chondrocytes,signaling pathway,and traditional Chinese medicine"in Chinese and English,respectively.The search time was from the inception of each database to March 2023.A total of 61 articles were included according to the inclusion and exclusion criteria. RESULTS AND CONCLUSION:The pathogenesis of knee osteoarthritis involves a complex molecular regulatory network,including aberrant expression of lncRNAs and miRNAs in cartilage tissues,which may lead to apoptosis of chondrocytes,degradation of cartilage extracellular matrix,and production of large amounts of pro-inflammatory cytokines.These changes interact with each other to cause degeneration of articular cartilage and progression of osteoarthritis.Therefore,further in-depth studies are needed to reveal the fine mechanisms of the molecular regulatory network.The mechanism of traditional Chinese medicine in the treatment of osteoarthritis mainly focuses on regulating the expression of lncRNA and miRNA,thereby alleviating chondrocyte apoptosis and extracellular matrix degradation,promoting cell proliferation,and slowing down the development of osteoarthritis.

Expression and Clinical Significance of Serum lncRNA H19,CHAST in Patients with Coronary Heart Disease / 医学研究杂志

Objective To investigate the expression level and clinical significance of serum long noncoding RNA(lncRNA)H19,CHAST in patients with coronary heart disease(CHD).Methods A total of 135 patients with CHD who were admitted to the Department of Cardiovascular Medicine of Lianyungang Second People's Hospital from June 2021 to May 2022 were selected,including 76 cases of a-cute coronary syndrome and 59 cases of chronic coronary syndrome.We selected 62 patients in the control group who were hospitalized and excluded from CHD by coronary angiography.Real-time fluorescent quantitative PCR(RT-PCR)was used to detect the relative expres-sion level of lncRNA H19 and lncRNA CHAST in the circulating serum of the subjects.Meanwhile,the high sensitivity C-reactive pro-tein(hs-CRP)、tumor necrosis factor-α(TNF-α)、interleukin-6(IL-6)、low density lipoprotein(LDL)、high density lipoprotein(HDL)and other related indicators of each group were collected.The receiver operating characteristic(ROC)curve was used to analyze the clinical diagnostic value of circulating serum lncRNA H19 and lncRNA CHAST expression level in the diagnosis of CHD.Pearson cor-relation coefficient was used to analyze the correlation between serum lncRNA H19,lncRNA CHAST level,and other related indicators.Multivariate Logistic regression was used to analyze the risk factors of CHD.Results The serum expression levels of lncRNA H19 and ln-cRNA CHAST in the CHD group were higher than those in the control group(P＜0.05).The serum levels of hs-CRP、TNF-α、IL-6、and LDL in the experimental group were higher than those in the control group(P＜0.05),and the serum levels of HDL were lower than those in the control group(P＜0.05).The expression levels of lncRNA H19 and lncRNA CHAST were positively correlated with hs-CRP,IL-6 and LDL,and were negatively correlated with HDL.lncRNA H19 and lncRNA CHAST are risk factors for CHD,which may have a higher value in the diagnosis of CHD.Conclusion lncRNA H19 and lncRNA CHAST are risk factors for CHD and can be used as serum indicators for the diagnosis of CHD,and the combination of the two has a higher value in the diagnosis of CHD.In addition,the ex-pression levels of both were correlated with the levels of inflammatory factors and apolipoproteins.

Long non-coding RNA AW112010 improves insulin resistance in adipocytes of aging mice through the miR-204/POU2F2 axis / 中华内分泌代谢杂志

Objective:To investigate whether long non-coding RNA(lncRNA) AW112010 can improve insulin resistance in aging adipocytes through the miR-204/POU2F2 signaling pathway.Methods:In vivo experiment: C57BL/6 mice were divided into young control group(4 months old) and aging model group(18 months old) based on body weight. The expression levels of AW112010, miR-204-5p, POU2F2, aging related indicators(p16, p21), and insulin signaling pathway genes [insulin receptor(INSR), insulin receptor substrate 1(IRS1), phosphatidylinositol kinase(PI3K), protein kinase B(AKT)] in epididymal adipose tissue were detected using real-time fluorescence quantitative PCR(RT-qPCR) and Western blotting. In vitro experiment: Using adriamycin(ADR) to induce 3T3-L1 aging adipocyte model, β-gal staining was used to observe cellular senescence, and miR-204 inhibitor and miR-204 mimic small interfering RNA were successfully constructed and transfected into 3T3-L1 adipocytes. Results:RT-qPCR and Western blot results showed that compared with the young group, the expression of AW112010 in the adipose tissue of aging mice was increased, while the expression of miR-204-5p was decreased. The expressions of POU2F2, p16, and p21 in the adipose tissue of aging mice were increased, while the expressions of INSR, IRS1, PI3K, GLUT4 mRNA and protein were decreased. The β-gal stainging results showed that the number of 3T3-L1 senescent adipocytes induced by ADR was significantly increased, and the expression levels of AW112010, POU2F2, p16, and p21 in ADR-induced senescent adipocytes were increased compared with the control group, while the expression levels of miR-204-5p, INSR, IRS1, PI3K, GLUT4 were decreased, and remaining glucose in the culture medium was increased. Compared with control, overexpression of miR-204 resulted in decreased expressions of aging indicators p16, p21, and target gene POU2F2 while the expressions of INSR and GLUT4 were increased.Conclusion:Upregulation of lncRNA AW112010 in adipocytes of aging mice may induce insulin resistance by targeting miR-204-5p/POU2F2/IRS1.

Expression profile analysis of long non-coding RNAs in hippocampus of alcohol-dependent mice induced by double-bottle selective drinking / 中国生物制品学杂志

@#Objective To analyze the expression profiles of long non-coding RNAs(lncRNA)in hippocampus of alcoholdependent mice induced by double-bottle selective drinking.Methods The alcohol-dependent mouse model was established by double-bottle selective drinking method,and the control group was set up(drinking water). Three male mice with alcohol preference more than 60% and alcohol consumption more than 10 g/(kg·24 h)in alcohol group and random three male mice in control group were selected,of which bilateral hippocampal brain tissues were isolated and stored in liquid nitrogen. LncRNA and mRNA of mouse hippocampal brain tissue RNA samples were sequenced by using Agilent-084388 microarray,and the differential expression of lncRNA in samples was detected by using ncRNA microarray. The biological processes and signaling pathways involved in differential expression of lncRNA were clustered and enriched by Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis. Pearson correlation analysis was used to predict the coding genes co-expressed by each differentially expressed lncRNA. Hypergeometric distribution test was used to calculate the significance of differential gene enrichment in each corresponding transcription factor item,and Cytoscape software was used to draw a visual network diagram.Results Compared with the control group,totally 855 lncRNAs(FC ≥ 2. 0,P < 0. 05)were differentially expressed in the hippocampus of mice in alcohol group,of which 337 lncRNAs were up-regulated significantly,with NONMMUT025786.2 and NONMMUT072246.2 being the most up-regulated,and 518 significant downward adjustments were observed,with the largest downward adjustments being NONMMUT113098.1 and NONMMUT076455.1. There were 361 mRNAs differentially expressed in the two groups(FC ≥ 2. 0,P < 0. 05)with 271 mRNAs up-regulated significantly and 90 significant downward adjustments,among which,the most obvious up-regulated were Upf3b and Zfp943,and Adamts 13 and Ift 27 showed the largest downward adjustments. The differential expression of lncRNAs was most obvious in the positive regulation of cell surface,GTPase activity and cell vesicle transport;The main signaling pathways involved were propanoate metabolism,taurine metabolism,extracellular matrix receptor interaction and AMPK signaling pathway. The most abundant transcription factors were FOXL1 and LHX3,with 25 and 21 corresponding co-expressed genes,respectively.Conclusion Through high-throughput gene expression profile microarray analysis,the possible key regulatory sites of lncRNAs and mRNAs were obtained,which provided experimental basis for research of the molecular mechanism of alcohol dependence in the hippocampus.

Transcription Factor ETS1 Promotes Glioma Cell Growth by Activating LncRNA XIST / 肿瘤防治研究

Objective To explore the biological function and downstream mechanism of ETS1 in glioma. Methods Bioinformatics and immunohistochemistry were used to analyze the differential expression characteristics of ETS1 in gliomas; qRT-PCR was employed to detect the expression level of ETS1 mRNA and lncRNA X-inactive specific transcript (XIST). CCK-8 and 5-ethyl-2′-deoxyuridine experiments were conducted to detect cell growth. Western blot was used to detect the expression of apoptosis-related proteins (Bax, Bak, Bcl-2). PROMO database was utilized to predict the binding sites between ETS1 and XIST promoter. Dual-luciferase reporter gene assay and chromatin immunoprecipitation-quantitative polymerase chain reaction assays were performed to verify the binding relationship between ETS1 and the XIST promoter region. cBioPortal database was used to analyze the correlation between the expression of ETS1 mRNA and XIST in glioma tissues. Results The expression levels of ETS1 mRNA and protein were significantly upregulated in glioma (P<0.05). The depletion of ETS1 significantly inhibited the proliferation of glioma cells and promoted cell apoptosis (P<0.05). ETS1 could target and bind with the XIST promoter and promote the expression of XIST (P<0.05). The overexpression of XIST reversed the effects of ETS1 on the proliferation of glioma cells and the promotion of cell apoptosis (P<0.05). Conclusion ETS1 is highly expressed in glioma tissues. It could promote the expression of lncRNA XIST, boost the proliferation of glioma cells, and inhibit cell apoptosis.

Expression and significance of lncRNA SNHG25 in prostate cancer based on the TCGA database / 现代泌尿外科杂志

【Objective】 To analyze the expression of lncRNA SNHG25 in prostate cancer and its significance, so as to explore the biomarkers and potential therapeutic targets for the diagnosis and prognosis of this disease. 【Methods】 Based on the TCGA database, differential, survival, and clinical correlation analyses of SNHG25 were performed.SNHG25 expression in prostate cancer was analyzed in the UALCAN database to determine its relationship with the clinical and pathological characteristics.The lncRNA-miRNA-mRNA correlation analysis was performed.The relevant ceRNA regulatory network was constructed.Prostate cancer samples were divided into high and low SNHG25 expression groups, and differential SNHG25 related genes were filtered and then enriched. 【Results】 SNHG25 expression was significantly upregulated in prostate cancer specimens compared to normal prostate specimens (P0.05).Regulatory networks of SNHG25/miR-330-3p/DLX1 and RPL22L1 were constructed. 【Conclusion】 SNHG25 is highly expressed in prostate cancer tissues and correlated with poor prognosis.SNHG25 expression does not significantly correlate with age, T-stage, N-stage, and Gleason score.SNHG25/miR-330-3p/DLX1 and RPL22L1 regulatory networks may play an important role in the development of prostate cancer.SNHG25 may become a biomarker and potential therapeutic target for prostate cancer.

Analysis of the expression and clinical significance of prostate cancer tissue-specific lncRNAs based on bioinformatics databases / 现代泌尿外科杂志

【Objective】 To explore the expression and clinical significance of prostate cancer tissue-specific lncRNAs. 【Methods】 The gene differences of 492 prostate cancer tissues and 152 adjacent tissues in TCGA and GEO genomic databases were analyzed with bioinformatics methods. A total of 5 lncRNAs were screened out, and their specificity in prostate tissues and impact on the prognosis of patients were analyzed. 【Results】 The 5 lncRNAs included PCAT14, PCA3, CTBP1-AS, DRAIC, and GPC5-AS1. PCAT14 and PCA3 were specifically expressed in prostate cancer tissues, and elevated expression was related to the prognosis. Moreover, they were well correlated with prostate cancer-specific antigens such as KLK3, AMACR, SLC45A3, and so on. GO function enrichment analysis and KEGG pathway enrichment analysis showed that the differential expression of PCA3 was closely related to phagocytosis, cell recognition, defense response to bacteria, immunoglobulin complex, Golgi apparatus, antigen binding, chemokine receptor binding, white matter digestion and absorption, renin-angiotensin system and other signaling pathways, while the differential expression of PCAT14 was closely related to the activity of Golgi apparatus and ion channels, renin secretion, cAMP signaling pathway, and gonadotropin secretion-related signaling pathway. 【Conclusion】 PCA3 and PCAT14 are specifically expressed in prostate cancer tissues, not in normal tissues, which can be used as potential indicators for the diagnosis of prostate cancer.

Intervention Mechanism of Biejiajian Wan on Primary Liver Cancer by Regulating lncRNA SNHG5/miRNA-26a-5p/GSK-3β Signal Axis / 中国实验方剂学杂志

ObjectiveTo investigate the mechanism of Biejiajian Wan in the intervention of primary liver cancer based on long non-coding RNA SNHG5 （lncRNA SNHG5）/micro RNA-26a-5p （miRNA-26a-5p）/glycogen synthase kinase-3β （GSK-3β） signal axis. MethodDouble luciferase reporting assay was used to verify the targeted interaction between lncRNA SNHG5 and miRNA-26a-5p， miRNA-26a-5p， and GSK-3β in HepG2 cells. Nude-mouse transplanted tumor model of human HepG2 were established and randomly divided into model group， Biejiajian Wan low-dose group （0.5 g·kg-1）， medium-dose group （1.0 g·kg-1）， and high-dose group （2.0 g·kg-1）， and sorafenib group （100 mg·kg-1）， with 10 mice in each group. The mice were given intragastric administration of normal saline or drug for 28 days， and the tumor volume was measured at different time. Hematoxylin-eosin （HE） staining was used to observe the histological changes of tumors. The nucleic acid levels of lncRNA SNHG5， miRNA-26a-5p， GSK-3β， and β-catenin mPNA in tumor tissue were detected by real-time quantitative polymerase chain reaction （Real-time PCR）. The protein expression levels of GSK-3β and β-catenin in tumor tissue were detected by western blot. ResultCompared with the SNHG5-WT （wild type） + miRNA NC （negative control） group， the relative luciferase activities of the SNHG5-WT + miRNA-26a-5p mimic group were decreased （P<0.05）. Compared with the GSK-3β-WT + miRNA NC group， the relative luciferase activity of the GSK-3β-WT + miRNA-26a-5p mimic group was decreased （P<0.05）. Compared with the model group， the tumor volume of Biejiajian Wan low-dose， medium-dose， and high-dose groups was significantly decreased （P<0.05， P<0.01）. Compared with the model group， the cells in the tumor tissue of nude mice in each dose group of Biejiajian Wan were sparsely arranged with necrocytosis， which showed concentration-dependent changes. Compared with the model group， the expression levels of lncRNA SNHG5， GSK-3β， and β-catenin were decreased （P<0.05， P<0.01）， while the expression of miRNA-26a-5p was increased in each dose group of Biejiajian Wan （P<0.05， P<0.01）. Compared with the model group， the protein expression levels of GSK-3β and β-catenin were decreased in each dose group of Biejiajian Wan （P<0.05， P<0.01）. ConclusionBiejiajian Wan may affect the necrosis of liver cancer cells through lncRNA SNHG5/miRNA-26a-5p/GSK-3β signal axis and thus play an anti-tumor role. This research will provide more theoretical basis for the clinical application of Biejiajian Wan.