【作者】 王超；

【导师】 杨小淮；

【作者基本信息】 蚌埠医学院 ， 外科学（专业学位）， 2021， 硕士

【摘要】 背景:膀胱癌(bladder cancer)是泌尿系统最常见的恶性肿瘤之一,发病率逐年增加,然而,膀胱癌在机体中的发生发展分子机制仍然未阐明。通过生物信息学分析发现miR-204-5p与膀胱癌的进展密切相关,研究证实miR-204-5p能够调节基因的表达,为阐明膀胱癌发生发展机制提供基础。目的:应用多种细胞核分子生物学技术证明miR-204-5p在膀胱癌中的作用,并明确miR-204-5p通过调控基因促进膀胱癌细胞迁移、侵袭的分子机制。方法:从TCGA官网(https://portal.gdc.cancer.gov/)下载膀胱癌(BLCA)437例miRNA rawdata(Datatype:Isoform Expression Quantification,Workflow Type:BCGSC miRNA Profiling)及相应的临床病理特征及生存信息。正常样本及各临床信息记录不齐备者被剔除。最终395例BLCA病人被纳入后续分析,通过生物信息学分析发现miR-204-5p与膀胱癌的进展密切相关,构建miR-204-5p稳定过表达和沉默的膀胱癌细胞株,观察地miR-204-5p过表达和沉默后膀胱癌细胞迁移和侵袭能力的改变,证实miR-204-5p对膀胱癌细胞迁移和侵袭能力的影响。结果:1.Rx64 3.6.1 software分析数据,通过生物信息学分析发现miR-204-5p与膀胱癌的进展密切相关。2.hsa-miR-204-5p在BLCA中具有差异表达,差异表达与不同的临床病理特征有统计学差异,且高表达的OS更差(P<0.05)。3.hsa-miR-204-5p在BLCA中cox单因素分析和cox多因素分析中具有统计学意义(P<0.05)。4.hsa-miR-204-5p在三个预测数据库中(>2个预测阳性的)共有866个靶基因,利用866个靶基因做GO及KEGG富集,发现与Hippo、Wnt等信号通路有关(P<0.05)。5.划痕和transwell实验研究证实miR-204-5p过表达后膀胱癌细胞迁移和侵袭能力增强,沉默后膀胱癌细胞迁移和侵袭能力减弱,差异具有统计学意义(P<0.05)。结论:hsa-miR-204-5p在BLCA中差异表达,差异表达与不同的临床病理特息息相关,miR-204-5p过表达后膀胱癌细胞迁移和侵袭能力增强,miR-204-5p沉默后膀胱癌细胞迁移和侵袭能力减弱,说明miR-204-5p通过调控作用从而影响膀胱癌细胞迁移、侵袭的能力。更多还原

【Abstract】 Background:Bladder cancer is one of the most common malignant tumors of the urinary system.The incidence of bladder cancer is increasing year by year.However,the molecular mechanism of the development and progression of bladder cancer in the organism has not been fully elucidated.Bioinformatics analysis revealed that miR-204-5p is closely related to the progression of bladder cancer,and studies have confirmed that miR-204-5p can regulate gene expression,providing a basis for elucidating the mechanism of the development and progression of bladder cancer.Objective:The role of miR-204-5p in bladder cancer was demonstrated by a variety of nuclear and molecular biology techniques,and the molecular mechanism of miR-204-5p promoting the migration and invasion of bladder cancer cells by regulating genes was clarified.Methods:From the TCGAwebsite(https://portal.gdc.cancer.gov/)to download(BLCA),437 cases of bladdercancer ofmicrornas rawdata(datatype: Isoform Expression Quantification,Workflowtype: BCGSC miRNA Profiling)and thecorresponding clinical pathological characteristics andsurvivalinformation.Normal samples and incomplete clinicalrecords were excluded.395 BLCApatients eventually beincorporated inthesubsequent analysis,through bioinformaticsanalysis found that miR-204-5 p is closely related to the progress of thebladder,build miR-204-5 p stable expression and thesilenceof the bladdercancer cell lines,observation of miR-204-5 p afterexpressing andsilencebladder cancercell migration and the changeof the invasiveability,confirmed that miR-204-5 p effects on bladder cancercell migration and invasion ability.Results:1.RX64 3.6.1 Software analyzed the data,and bioinformatics analysis showed that miR-204-5p was closelyrelated to theprogression of bladdercancer.2.Hsa-miR-204-5p was differentially expressed in BLCA,and the differentially expressed miR-204-5p was statistically different with different clinicopathological features,and the OS with high expression was worse(P < 0.05).3.Hsa-miR-204-5p showed statistical significance in COX univariate analysis and COX multivariate analysis in BLCA(P < 0.05).4.There were a total of 866 target genes of hsa-miR-204-5p in the three prediction databases(those with more than 2 predicted positive genes),and 866 target genes were enriched in GO and KEGG,which were found to be related to HIPPO,Wnt and other signaling pathways(P< 0.05).5.The scratch and Transwell experiments confirmed that the migration and invasion ability of bladder cancer cells were enhanced after the overexpression of miR-204-5p,but decreased afterthe silencing,withstatistical significance(P < 0.05).Conclusion:HSA-miR-204-5p is differentially expressed in BLCA,which is closely related to different clinicopathological characteristics.After overexpression of miR-204-5p,the migration and invasion ability of bladder cancer cells is enhanced,while after silencing of miR-204-5p,the migration and invasion ability of bladder cancer cells is weakened,suggesting that miR-204-5p affects themigration and invasion ability of bladdercancer cells through regulation.更多还原