miR-30a-3p通过靶向调控BAFF在肾癌细胞迁移和血管生成能力中的作用研究  

The Role of miR-30a-3p in the Migration and Angiogenesis Ability of Renal Cancer Cells through Targeted Regulation of BAFF

作　　者：威力江·赛买提[1] 马军 王玉杰[1] WEILIJIANG·Saimaiti;MA Jun;WANG Yujie(The First Affiliated Hospital of Xinjiang Medical University,Xinjiang Urumqi 830011,China)

机构地区：[1]新疆医科大学第一附属医院,新疆乌鲁木齐830011

出　　处：《河北医学》2023年第4期529-535,共7页Hebei Medicine

基　　金：新疆维吾尔自治区自然科学基金项目,(编号:2020D01C261)。

摘　　要：目的:研究微小RNA(miR)-30a-3p对肾细胞癌(RCC)细胞的转移和血管生成表型的影响。方法:结合癌症基因组图谱(TCGA)数据库提供的RCC中miR-30a-3p表达数据分析miR-30a-3p在RCC细胞系中的表达特征。结合生物信息学网站预测miR-30a-3p与B细胞激活因子(BAFF)3’非翻译区(3’-UTR)的结合。将RCC细胞系ACHN和786O分为LV-miR-30a-3p组[感染过表达miR-30a-3p的慢病毒载体]、LV-NC组[感染慢病毒载体阴性对照]、LV-miR-30a-3p+pcDNA-BAFF组[联合感染LV-miR-30a-3p和过表达BAFF的载体(pcDNA-BAFF)]以及LV-miR-30a-3p+pcDNA-EV组[联合感染LV-miR-30a-3p和空载体(pcDNA-EV)]。Transwell迁移小室法检测细胞的迁移能力,小管形成实验检测细胞血管生成能力。qPCR检测miR-30a-3p的表达。Western blot检测BAFF、促血管生成蛋白血管内皮生长因子A(VEGFA)的蛋白表达。双荧光素酶基因报告实验检测ACHN细胞中的miR-30a-3p对BAFF的靶向调控作用。结果:分析TCGA数据库中数据,与Normal组比,Cancer组的miR-30a-3p表达都显著降低(P<0.05);与HK-2细胞比,ACHN和786O细胞系中miR-30a-3p的表达都明显降低(均P<0.05)。与LV-NC组比,LV-miR-30a-3p组miR-30a-3p表达增加(均P<0.05),LV-miR-30a-3p组的细胞迁移、小管生成、及BAFF和VEGFA表达均下调(均P<0.05)。经过生物信息学预测以及双荧光素酶报告基因实验验证发现BAFF是miR-30a-3p的靶基因。与LV-miR-30a-3p+pcDNA-EV组比较,LV-miR-30a-3p+pcDNA-BAFF组的细胞迁移、小管生成率及BAFF表达均上调(均P<0.05)。结论:miR-30a-3p通过靶向抑制BAFF抑制RCC细胞的迁移和血管生成能力。Objective:To investigate the effects of microRNA(miR)-30A-3p on migration and angiogenic phenotype of renal cell carcinoma(RCC)cells.Methods:The expression characteristics of miR-30a-3p in RCC cells were analyzed based on the expression data of miR-30a-3p in the Cancer Genome Atlas(TCGA)database.Predict the binding of miR-30a-3p to the 3'untranslated region(3'-UTR)of B cell activicting factor(BAFF)by bioinformatics websites.RCC cell lines ACHN and 786O were divided into LV-miR-30a-3p group[infected with lentiviral vector overexpressing miR-30a-3p],LV-NC group[infected with lentiviral vector negative control],LV-miR-30a-3p+pcDNA-BAFF group[co-infected with LV-miR-30a-3p and LV-Mir-30a-3p Overexpressing BAFF vector(pcDNA-BAFF)]and LV-miR-30a-3p+pcDNA-EV group[co-infected with LV-miR-30a-3p and empty vector(pcDNA-EV)].Transwell migration chamber assay was used to detect cell migration ability,and tubule formation assay was used to detect cell angiogenesis ability.The expression of miR-30a-3p was detected by qPCR.The protein expressions of BAFF and vascular endothelial growth factor(VEGFA)were detected by Western blot.Targeting regulation of miR-30a-3p against BAFF in ACHN cells was detected by dual luciferase gene reporting assay.Results:Analysis of the data in the TCGA database showed that miR-30a-3p expression was significantly lower in the Cancer group compared to the Normal group(P<0.05).Compared with HK-2 cells,the expression of miR-30a-3p in ACHN and 786O cell lines was significantly decreased(both P<0.05).Compared with the LV-NC group,the expression of miR-30a-3p was increased in the LV-miR-30a-3p group(all P<0.05),while the cell mobility,tubule formation rate,and the expressions of BAFF and VEGFA were down-regulated in the LV-miR-30a-3p group(all P<0.05).According to bioinformatics prediction and double luciferase reporter assay,BAFF was found to be the target gene of miR-30a-3p.Compared with the LV-miR-30a-3p+pcDNA-EV group,cell mobility,tubule formation rate and BAFF expression in the LV-miR-30a-3p+pcDNA-B

关 键 词：微小RNA-30a-3p 肾细胞癌 B细胞激活因子 细胞迁移 血管生成 

分 类 号：R737.11[医药卫生—肿瘤]