过表达miR-330-3p通过BMI1对黑色素瘤A375细胞增殖、侵袭及凋亡的影响

The effect of overexpression of miR-330-3p on the proliferation invasion and apoptosis of melanoma A375 cells via BMI1

目的探讨过表达miR-330-3p对黑色素瘤A375细胞增殖、侵袭及凋亡的影响。方法采用miR-330-3p模拟物转染黑色素瘤A375细胞,设置空白组(未进行转染)、miR-330-3p正常对照组(转染无义序列)、miR-330-3p模拟物转染组(转染miR-330-3p模拟物),采用CCK-8法检测各组A375细胞的增殖,Transwell检测各组细胞侵袭能力,流式细胞仪检测各组细胞凋亡,Western blot检测各组细胞BMI1蛋白表达,应用Targetscan和双荧光素酶报告基因实验检测BMI1是否为miR-330-3p的靶基因。结果miR-330-3p过表达后,A375细胞的增殖和侵袭能力下降,细胞凋亡率上升,BMI1蛋白表达水平下降,生物信息学分析及双荧光素酶实验证实BMI1为miR-330-3p的靶基因。结论过表达miR-330-3p抑制A375细胞增殖和侵袭,促进其凋亡,与下调BMI1的表达相关。

Objective To investigate the effect of overexpression of miR-330-3 p on the proliferation,invasion and apoptosis of melanoma cells.Methods miR-330-3 p mimics was transfected into A375 cells,the experiment was divided into:blank group(not transfected),miR-330-3 p normal control group(transfected nonsense sequence)and miR-330-3 p mimics group(transfected miR-330-3 p mimics).CCK-8 was used to detect the proliferation,transwell was used to detect the invasion,flow cytometry was used to detect the apoptosis,Western blot was used to detect the expression of BMI1 protein,and the target scan and double luciferase reporter gene experiment was used to verify whether BMI1 is the target gene of miR-330-3 p.Results The overexpression of miR-330-3 p in A375 cells,down-regulated BMI1 expression and the proliferation and invasion abilities,and promoted the apoptosis of A375 cells.Bioinformatics analysis and dual luciferase reporter gene confirmed that BMI1 was the target gene of miR-330-3 p.Conclusion The overexpression of miR-330-3 p inhibits the proliferation and invasion,promotes apoptosis in A375 cells,which is related to the downregulation of BMI1 expression.