miR-509-5p靶向ACLY调控人黑色素瘤细胞的增殖和凋亡  

Effects of miR-509-5p targeting ACLY on the proliferation and apoptosis of human melanoma cells

作　　者：李二龙[1] 张皓义 韩成龙 谢林伸 LI Erlong;ZHANG Haoyi;HAN Chenglong(Department of Dermatosis and Syphilology,West China Hospital of Sichuan University,Sicuan,Chengdu 610041,China)

机构地区：[1]四川大学华西医院皮肤性病科,成都市610041 [2]四川大学华西公共卫生学院,华西第四医院职业病科,成都市610041

出　　处：《河北医药》2021年第1期39-43,共5页Hebei Medical Journal

基　　金：成都市科技局科技项目(编号:2015-HM01-00349-SF)。

摘　　要：目的探讨miR-509-5p靶向调控ACLY表达对人黑色素瘤细胞增殖和凋亡的影响。方法qRT-PCR检测人表皮黑色素细胞HEMn-LP和3种人黑色素瘤细胞(SK-MEL-2、UACC903和A375)中miR-509-5p和ACLY mRNA的表达水平。以A375细胞为研究对象,构建过表达miR-509-5p的A375细胞株,MTT法检测细胞活力,流式细胞术检测细胞凋亡,Western blot检测CyclinD1、CDK2、CDK1、Bcl-2、Bax和cleaved caspase-3蛋白的表达。双荧光素酶报告基因实验和western blot验证miR-509-5p和ACLY的靶向关系。结果与人表皮黑色素细胞HEMn-LP相比,3种人黑色素瘤细胞中miR-509-5p的表达显著下调,ACLY mRNA的表达显著上调。过表达miR-509-5p可抑制CDK2、CDK1、CyclinD1和Bcl-2蛋白的表达,促进Bax和cleaved caspase-3蛋白的表达,抑制A375细胞增殖,促进细胞凋亡。ACLY是miR-509-5p的靶基因,miR-509-5p可负性调控ACLY的表达。过表达ACLY可部分逆转miR-509-5p过表达对A375细胞增殖和凋亡的影响。结论miR-509-5p靶向下调ACLY表达可抑制人黑色素瘤细胞增殖,促进细胞凋亡。Objective To investigate the effects of miR-509-5p targeting ACLY expression on the proliferation and apoptosis of human melanoma cells.Methods The qRT-PCR was used to detect the expression levels of miR-509-5p and ACLY mRNA in human epidermal melanoma cell HEMn-LP and three human melanoma cells(SK-MEL-2,UACC903 and A375).A375 cells were enrolled as research subjects,and the A375 cell line overexpressing miR-509-5p was constructed.The cell viability was detected by MTT,cell apoptosis was detected by flow cytometry,and expression levels of CyclinD1,CDK2,CDK1,Bcl-2,Bax and cleaved caspase-3 protein were detected by Western Blot.The dual luciferase reporter gene experiment and western blot were used to verify the targeting relationship between miR-509-5p and ACLY.Results As compared with those in human epidermal melanoma cell HEMn-LP,the expression levels of miR-509-5p in the three human melanoma cells were significantly down-regulated,however,the expression levels of ACLY mRNA were significantly up-regulated.The overexpression of miR-509-5p could inhibit the expression levels of CDK2,CDK1,CyclinD1 and Bcl-2 protein,however,which could promote the expression levels of Bax and cleaved caspase-3 protein,and inhibit the proliferation of A375 cells,and promote the cell apoptosis.ACLY was the target gene of miR-509-5p that could negatively regulate the expression of ACLY.Moreover the overexpression of ACLY could partially reverse the effects of miR-509-5p overexpression on the proliferation and apoptosis of A375 cells.Conclusion The miR-509-5p can inhibit the proliferation and promote the apoptosis of human melanoma cells by targetedly down-regulating ACLY expression.

关 键 词：人黑色素瘤 miR-509-5p ATP柠檬酸裂解酶 细胞增殖 凋亡 

分 类 号：R739.5[医药卫生—肿瘤]