摘要
目的 :探讨微RNA-378(micro RNA-378,mi R-378)对肾癌细胞体外增殖和迁移能力的影响,以及其可能的作用机制。方法 :利用TCGA(e Cancer Genome Atlas)数据库分析肾癌和正常肾上皮组织中mi R-378的表达情况。采用实时荧光定量PCR法检测人肾癌细胞系786-O、ACHN、769-P和Caki-1,以及人肾小管上皮细胞系HK2中mi R-378的表达情况。人工合成mi R-378模拟物(mimic)和抑制子(inhibitor),并将它们分别转染肾癌786-O和Caki-1细胞,然后分别采用CCK-8法、Transwell小室法和划痕愈合实验检测mi R-378对肾癌细胞增殖和迁移的影响。通过生物信息学方法分析mi R-378的靶基因,并采用实时荧光定量PCR及蛋白质印迹法进行验证。结果 :肾癌组织和肾癌细胞系中mi R-378表达均明显上调(P值均<0.001)。mi R-378 mimic转染后,肾癌786-O细胞的体外增殖(P<0.01)和迁移(P<0.001)能力均明显增强;而mi R-378 inhibitor转染后,肾癌Caki-1细胞的体外增殖(P<0.01)和迁移(P<0.001)能力均明显减弱。肾癌中mi R-378的靶基因可能是大型肿瘤抑制因子2(large tumor suppressor 2,LATS 2),肾癌786-O和Caki-1细胞中LATS2 m RNA和蛋白表达水平与mi R-378水平均负相关(P值均<0.01)。结论 :mi R-378过表达可增强人肾癌细胞的增殖和迁移能力,其机制可能与负调控LATS2表达有关。
Objective： To investigate the effects of micro RNA-378（mi R-378） on proliferation and migration of renal cancer cell lines in vitro, and to explore the possible mechanism.Methods： The expression of mi R-378 in renal cell carcinoma and normal renal epithelial tissues was analyzed using the datasets from The Cancer Genome Atlas（TCGA）. The expression of mi R-378 in human renal carcinoma cell lines 786-O, ACHN, 769-P and Caki-1 as well as human renal proximal tubular epithelial cell line HK2 was detected by real-time fluorescent quantitative PCR（RFQ-PCR）. Mi R-378 mimic and inhibitor were synthesized and then transfected into renal carcinoma 786-O and Caki-1 cells, respectively. The proliferation and migration of 786-O and Caki-1 cells were detected by CCK-8 assay, Transwell chamber assay and wound healing assay, respectively. The potential target gene of mi R-378 was screened by bioinformatics, and confirmed by RFQ-PCR and Western blotting in renal cancer cells, respectively. Results： The expression level of mi R-378 in renal cancer tissues and cell lines was significantly up-regulated as compared with that in normal renal epithelial tissues or normal renal epithelial HK2 cells（all P〈0.001）. After transfection with mi R-378 mimic, the proliferation（P〈0.01） and migration（P〈0.001） of 786-O cells were remarkably increased. In contrast, after transfection with mi R-378 inhibitors, the proliferation（P〈0.01） and migration（P〈0.001） of Caki-1 cells were remarkably suppressed. By bioinformatics assay, the large tumor suppressor 2（LATS 2） was identified as a target gene of mi R-378 in renal cancer. The m RNA and protein expression of LATS2 were negatively related with the expression of mi R-378 in renal cancer 786-O and Caki-1 cells（all P〈0.01）. Conclusion： Overexpression of mi R-378 may promote the proliferation and migration of renal cancer cells by negative targeting LATS 2 gene.
作者
安娜
王晓
周培杰
魏莲子
方煜翔
高维强
AN Na;WANG Xiao;ZHOU Peijie;WEI Lianzi;FANG Yuxiang;GAO Weiqiang(Renji-Med X Clinical Stem Cell Research Center,Renji Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200217,China)
出处
《肿瘤》
CAS
CSCD
北大核心
2018年第7期643-651,共9页
Tumor
基金
国家自然科学基金资助项目(编号:81630073)~~
