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 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.

Mechanism of lncRNA MIR4435-2HG targeting miR-376a-3p to regulate the biological behavior of cholangio-carcinoma cells / 局解手术学杂志

Objective To explore the effects of long non-coding RNA(lncRNA)MIR4435-2HG(MIR4435-2HG)on the proliferation,migration,invasion and apoptosis of cholangiocarcinoma cells and its regulatory effect on microRNA-376a-3p(miR-376a-3p).Methods qRT-PCR method was used to detect the expression of MIR4435-2HG and miR-376a-3p in human intrahepatic bile duct epithelial cells HIBEpic and human cholangiocarcinoma cells RBE.si-NC,si-MIR4435-2HG,miR-NC,miR-376a-3p mimics,si-MIR4435-2HG and anti-miR-NC,and si-MIR4435-2HG and anti-miR-376a-3p were transfected into RBE cells,respectively,as the si-NC group,the si-MIR4435-2HG group,the miR-NC group,the miR-376a-3p group,the si-MIR4435-2HG+anti-miR-NC group,the si-MIR4435-2HG+ anti-miR-376a-3p group.MTT method,Transwell chamber method and flow cytometry were used to detect cell proliferation,migration,invasion and apoptosis;dual luciferase reporter gene assay was used to verify the targeting relationship between MIR4435-2HG and miR-376a-3p.Western blot was used to detect the expression of related proteins.Results The expression of MIR4435-2HG was increased in RBE cells,while the expression of miR-376a-3p was decreased(P<0.05).Compared with the si-NC group,the MIR4435-2HG expression,cell viability,and protein levels of CyclinD1,MMP-2,MMP-9 in the si-MIR4435-2HG group were reduced(P<0.05),the numbers of migrating and invading cells were reduced(P<0.05),while the MIR4435-2HG expression and apoptosis rate were increased(P<0.05).Compared with the miR-NC group,the cell viability and protein levels of CyclinD1,MMP-2,MMP-9 in the miR-376a-3p group were decreased(P<0.05),the numbers of migrating and invading cells were decreased(P<0.05),while the MIR4435-2HG expression and apoptosis rate were increased(P<0.05).MIR4435-2HG was of targeted regulation on miR-376a-3p.Compared with the si-MIR4435-2HG+ anti-miR-NC group,the cell viability and protein levels of CyclinD1,MMP-2,MMP-9 in the si-MIR4435-2HG+anti-miR-376a-3p group were increased(P<0.05),the numbers of migrating and invading cells were increased(P<0.05),while the MIR4435-2HG expression and apoptosis rate were decreased(P<0.05).Conclusion Knockdown of MIR4435-2HG can inhibit the proliferation,migration,invasion and induce apoptosis of RBE cells by targeting miR-376a-3p.

Regulation and mechanism of lncRNA H19 in autoimmune diseases / 中国免疫学杂志

Long non-coding RNA(lncRNA)is a type of non-protein-coding RNA with a length of at least 200 nucleotides.Ex-isting evidence shows that lncRNA affects the growth and development of humans and the occurrence of diseases through various mech-anisms,and plays an irreplaceable role in the differentiation and activation of immune cells and the development process of autoim-mune diseases.In recent years,lncRNA H19 has been found to play a unique regulatory function in pathological processes related to autoimmune diseases,such as inflammation and fibrosis.Therefore,the molecular mechanism involved in H19 is expected to become a potential target for the treatment of autoimmune diseases.In this review,we discussed the related mechanisms of H19 involved in the pathogenesis of autoimmune diseases based on the known functions of H19 and summarized the research results of H19 in several com-mon autoimmune diseases.

Platelet lncRNA as a biomarker for early screening of lung adenocarcinoma: a preliminary study / 中国输血杂志

【Objective】 To explore the diagnostic value of platelet long non-coding RNA (lncRNA) as a biomarker for early screening of lung adenocarcinoma (LUAD). 【Methods】 The GSE183635 and GSE89843 datasets, which contained the platelet transcriptome of LUAD and healthy controls, were used for differential analysis, and the intersection of the differentially expressed lncRNA(DElncRNA) of the two datasets was taken. The expression levels of DElncRNA in LUAD tissues and normal control tissues were analyzed using GEPIA2. The expression levels of LINC01088 in platelets of 51 healthy controls and 54 LUAD patients were detected by quantitative Real-time PCR (qRT-PCR), and the diagnostic ability of each index was evaluated by ROC curve. 【Results】 8 DElncRNAs and 1 265 DElncRNAs were obtained from GSE183635 and GSE89843 datasets, respectively. The key DElncRNA LINC01088 was selected after intersection. GEPIA2 analysis showed that the expression level of LINC01088 in LUAD tissues was lower than that in normal lung tissues (P<0.05). Platelet LINC01088 was significantly downregulated in patients with LUAD and early-stage LUAD than in healthy controls(P<0.001). The area under the curve (AUC) of platelet LINC01088 in the diagnosis of LUAD was 0.755, the sensitivity was 81.1%, and the specificity was 67.9%. The AUC for early LUAD diagnosis was 0.727, the sensitivity was 80.0%, and the specificity was 67.9%. The AUC of the combined diagnostic model composed of platelet LINC01088 and carcinoembryonic antigen (CEA) for LUAD diagnosis was 0.807, the sensitivity was 89.2%, and the specificity was 71.4%. The AUC for early LUAD was 0.770, the sensitivity was 86.7%, and the specificity was 71.4%. The combined diagnostic model of platelet LINC01088 and CEA was superior to CEA in the diagnosis of LUAD and early LUAD (Z=-2.288, -2.34, both P<0.05). 【Conclusion】 LINC01088 is down-regulated in platelets of LUAD patients. Platelet LINC01088 may be a biomarker for early screening and diagnosis of LUAD.

Research progress into the mechanisms of exosome immunoregulation of bacterial infection / 中国比较医学杂志

Exosomes are small endosomally derived extracellular vesicles with a lipid bilayer structure,and they contain substances,such as proteins,lipids,DNA,RNA,micro(mi)RNA,and long non-coding(lnc)RNA.Exosomes participate in pathogen recognition,antigen presentation,autophagy regulation,immune activation and immunosuppression in bacterial infections.Studies have shown that miRNA,lncRNA,and proteins in exosomes play important roles in regulating antibacterial reactions in organisms.We reviewed the immunomodulatory effects of exosomes on several intracellular and extracellular bacterial infections to provide a reference for those studying the interactions between exosomes and bacterial infections.

Effect and Potential Mechanism of Inhibition of Long Non-coding RNA MALAT1 on Glycolipipotoxicity-induced Endothelial Cell Dysfunction / 中国循环杂志

Objectives:To investigate the effect of inhibition of long non-coding RNA(lnc RNA)in human metastasis associated lung adenocarcinoma transcript 1(MALAT1)on glycolipitoxicity-induced human umbilical vein endothelial cell dysfunction. Methods:Human umbilical vein endothelial cells were treated with glucose and palmitic acid in vitro to establish the glycolipitoxic endothelial cell models.Following groups were examined:control group,high-glucose and high-fat group,high-glucose and high-fat + non-targeting RAN control group,high-glucose and high-lipid+MALAT1 siRNA group,and high-glucose and high-lipid+MAPK1 siRNA group.RT-qPCR was used to detect the mRNA expression of MALAT1 and MAPK1.Western blot was used to detect the expression levels of autophagy,mitochondrial fusion division,apoptosis,and pathway-related proteins.Immunofluorescence confocal localization was used to detect the fluorescence colocalization of autophagy and lysosome-related proteins.The number of autophagolysosomes in endothelial cells was observed by transmission electron microscopy.Mitochondrial probe staining was used to detect mitochondrial morphology,immunofluorescence was used to detect intracellular reactive oxygen species(ROS)production,flow cytometry was used to detect the apoptosis of cells in each group,cell proliferation and scratch assays were used to detect the proliferation and migration ability of cells in different groups at different time points.The angiogenesis was quantified by counting the number of new blood vessels in each group. Results:Compared with the control group,the expression of lncRNA MALAT1 mRNA and the expression of phosphorylated mito-activated protein kinase 1(p-MAPK1)were upregulated(both P＜0.05)and the expression of phosphorylated mammalian target protein(p-mTOR)was downregulated in the high-glucose and high-fat group and the high-sugar and high-fat control group(all P＜0.01).Compared with the high-glucose and high-fat non-targeting RNA control group,the expressions of microtubule-associated protein 1A/1B-light chain 3(LC3)and p62 were downregulated(P＜0.01,P＜0.05),LC3 and lysosome-associated membrane protein 2(LAMP2)protein co-localized positive fluorescence particles were increased(both P＜0.01),number of lysosomes were decreased,the expression of ROS was decreased(P＜0.01),the expression level of mitochondrial fusion protein optic nerve atrophin 1(OPA1)was increased(P＜0.05),the expressions of cleaved caspase-3 and BCL-2-related X protein(BAX)were decreased and BCL-2 was increased(all P＜0.05),cell proliferation,migration,and tube-forming ability were increased(all P＜0.01),and the expression of p-MAPK1 was decreased(P＜0.05)and p-mTOR expression was increased(both P＜0.05)in the high-glucose and high-lipid+si-MALAT1 group.Compared with the high-glucose and high-fat non-targeting RNA control group,the expression of p-MAPK1 in endothelial cells was decreased and the expression of p-mTOR was increased in the high-glucose and high-lipid+si-MAPK1 group(both P＜0.01). Conclusions:Inhibition of lncRNA MALAT1 expression can reduce the level of mitophagy in glycolipidotoxic environments,reduce apoptosis of endothelial cells and improve endothelial cell function,which may be related to the regulation of MAPK1/mTOR signaling pathway.

Study on the Mechanism of Yangjing Zhongyu Decoction in Regulating Primordial Follicle Initiation in Model Rats with Diminished Ovarian Reserve / 中国中医药信息杂志

Objective To study the mechanism of Yangjing Zhongyu Decoction in regulating the initiation of primordial follicles in model rats with diminished ovarian reserve(DOR)based on lncRNA.Methods Three-day-old female rats were selected and their ovaries were cultured in vitro.The blank group,model group,DHEA group and Yangjing Zhongyu Decoction high-,medium-and low-dosage groups were set.The DOR model was induced by triptolide,corresponding drug containing serum was given to culture respectively.HE staining was used to observe germ cells and follicles,Western blot was used for determining the expressions of AMH,BMP15,PTEN,MST,TGF-β1,p-Smad1 protein,RT-PCR was used to detected AMH,BMP15,PTEN,MST,LTCONS-00011173,TGF-β1,Smad1 mRNA expression.Results Compared with the blank group,the number of primordial and growing follicles in the model group rats decreased(P<0.05),the expression of AMH,BMP15,TGF-β1,p-Smad1 protein in ovarian tissue decreased(P<0.05),expressions of PTEN and MST proteins increased(P<0.05),AMH,BMP15,TGF-β1,Smad1 mRNA expression decreased(P<0.05),while the expressions of PTEN,MST,and LTCONS-00011173 mRNA increased(P<0.05).Compared with the model group,the DHEA group and Yangjing Zhongyu Decoction high-and medium-dosage groups showed an increase in the number of primordial and growing follicles(P<0.05),the expressions of AMH,BMP15,TGF-β1 and p-Smad1 protein in ovarian tissue increased(P<0.05),PTEN and MST protein expressions decreased(P<0.05),AMH,BMP15,TGF-β1,Smad1 mRNA expressions increased(P<0.05),while PTEN,MST,and LTCONS-00011173 mRNA expressions decreased(P<0.05).Conclusion Yangjing Zhongyu Decoction may mediate TGF-β1/Smad1 signaling pathway through LTCONS-00011173,regulating primordial follicle initiation in DOR model rats.

Effect and mechanism of lncRNA GACAT3 on the radiation resistance of gliomas by regulating the miR-497/YAP1 axis / 中华放射肿瘤学杂志

Objective:To investigate the role and mechanism of long non-coding RNA (lncRNA) gastric cancer associated transcript 3 (GACAT3) in glioma radioresistance.Methods:Real-time reverse transcription PCR (RT-qPCR) was used to detect the expression of lncRNA GACAT3 and miR-497 in human astrocyte NHA cells and glioma cells U251. NC-siRNA and GACAT3-siRNA were transfected into U251 cells, and the cells were treated with X-ray irradiation. Colony formation assay was used to detect the survival fraction of U251 cells. The apoptosis of U251 cells was detected by flow cytometry. Western blot was used to detect the expression of cysteine containing aspartate specific protease 3 (Caspase-3) in U251 cells. Bioinformatics software and dual luciferase reporter gene assay were used to predict and verify the targeting relationship between lncRNA GACAT3 and miR-497, and between miR-497 and Yes-associated protein 1 (YAP1), respectively. NC mimic, miR-497 mimic, GACAT3-siRNA and NC inhibitor, GACAT3-siRNA and miR-497 inhibitor were co-transfected into U251 cells. Colony formation assay, flow cytometry and Western blot were adopted to evaluate the effect of miR-497 overexpression and lncRNA GACAT3 on the radiosensitivity of U251 cells by regulating miR-497.Results:Compared with NHA cells, the expression of lncRNA GACAT3 in U251 cells was significantly up-regulated, and the expression of miR-497 in U251 cells was significantly down-regulated (both P<0.05). After knockdown of GACAT3, the survival fraction of irradiated U251 cells was significantly decreased, while the apoptosis rate and Caspase-3 protein expression were significantly increased (all P<0.05). lncRNA GACAT3 targeted and negatively regulated the expression of miR-497. Overexpression of miR-497 significantly reduced the survival fraction of U251 cells after irradiation, and increased the apoptosis rate and Caspase-3 protein expression. Inhibition of miR-497 significantly reversed the promoting effect of lncRNA GACAT3 knockdown on the radiosensitivity of U251 cells. miR-497 targeted and negatively regulated the expression of YAP1. Conclusion:Knockdown of lncRNA GACAT3 can enhance the radiosensitivity of glioma cells by regulating the miR-497/YAP1 axis.

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.

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.