摘要
目的探讨miR-520a-3p在肾癌细胞系中的表达水平及其对肾癌细胞增殖、迁移、侵袭和凋亡的影响。方法通过实时荧光定量聚合酶链式反应检测miR-520a-3p在正常肾小管上皮细胞HK2和肾癌细胞系786-O、A498、OS-RC-2及ACHN中的表达水平。选取miR-520a-3p表达水平最低的肾癌细胞系786-O和A498为实验对象,各分为两组:对照组(转染miR-NC)和实验组(转染miR-520a-3p模拟物)。采用CCK-8实验、Transwell迁移和侵袭实验、流式细胞术检测各组细胞增殖、迁移、侵袭和凋亡情况;采用Western blotting验证相关蛋白表达量变化。采用生物信息学软件预测miR-520a-3p的靶基因,双荧光素酶报告实验验证其靶向关系。结果相对于HK2细胞,miR-520a-3p在多种肾癌细胞系中表达明显降低(P<0.05),在786-O和A498细胞中降低最明显(P<0.05)。转染miR-520a-3p模拟物后,实验组的细胞增殖、迁移和侵袭能力明显降低,凋亡率增加(P<0.05),N-cadherin、Bcl2、Bcl3蛋白表达量明显降低,E-cadherin蛋白表达量明显增高。生物信息学软件预测miR-520a-3p与Bcl3存在可结合位点,双荧光素酶报告实验证实miR-520a-3p可直接靶向Bcl3。结论miR-520a-3p在肾癌细胞系中低表达,过表达miR-520a-3p可抑制肾癌细胞系786-O和A498增殖、迁移和侵袭能力,促进细胞凋亡。
Objective To investigate the expression level of miR-520a-3p in renal cancer cell lines and its effects on the proliferation,migration,invasion and apoptosis of renal cancer cells.Methods The expression of miR-520a-3p in normal renal tubular epithelial cells HK2 and renal cancer cell lines(786-O,A498,OS-RC-2 and ACHN)were detected by real-time quantitative polymerase chain reaction(qRT-PCR).We selected 786-O and A498 with the lowest expression of miR-520a-3p as experimental subjects.Each cell line was divided into two groups:the control group(transfected with miR-NC)and the experimental group(transfected with miR-520a-3p mimics).CCK-8 experiment,Transwell migration and invasion test,flow cytometry were used to detect cell proliferation,migration,invasion and apoptosis in each group.Western blotting was used to verify the expression level of related proteins.Bioinformatics was used to predict miR-520a-3p target genes,and dual luciferase reporter genes were used to verify their targeting relationship.Results Compared with HK2 cells,the expression of miR-520a-3p significantly reduced in a variety of renal cancer cell lines(P<0.05),especially in the 786-O and A498(P<0.05).After transfection of miR-520a-3p mimics,the cell proliferation,migration and invasion ability of the experimental group significantly reduced,and the rate of apoptosis increased(P<0.05),and the protein expression of N-cadherin,Bcl2 and Bcl3 decreased significantly,while the protein expression of E-cadherin increased significantly.There are binding sites between Bcl3 and miR-520a-3p,and dual luciferase reporter gene detection confirmed that Bcl3 is the target gene of miR-520a-3p.Conclusions The expression of miR-520a-3p significantly reduced in renal cancer cell lines.Over expression of miR-520a-3p can inhibit the proliferation,migration and invasion of renal cancer cell lines 786-O and A498,and induce apoptosis.
作者
翟官忠
刘泽林
王清华
柯帅
郭佳
ZHAI Guanzhong;LIU Zelin;WANG Qinghua;KE Shuai;GUO Jia(Department of Urology, Renmin Hospital of Wuhan University, Wuhan 430060, China)
出处
《现代泌尿生殖肿瘤杂志》
2021年第1期34-39,共6页
Journal of Contemporary Urologic and Reproductive Oncology
关键词
肾癌
miR-520a-3p
增殖
迁移
侵袭
凋亡
Renal cell carcinoma
miR-520a-3p
Proliferation
Migration
Invasion
Apoptosis
