摘要
目的:研究乙酰辅酶A合成酶2(acetyl-CoA synthase 2,ACSS2)对三阴性乳腺癌MDA-MB-468细胞增殖和凋亡的影响及可能的机制。方法:选用正常乳腺上皮MCF-10A细胞与三阴性乳腺癌MDA-MB-468细胞,采用实时荧光定量PCR(qRT-PCR)和蛋白质印迹法分别检测及比较两者ACSS2 mRNA和蛋白表达差异;设计并合成针对ACSS2的特异性干扰RNA(siRNA-ACSS2)及无序序列的阴性对照(siRNA-NC),分别瞬时转染MDA-MB-468细胞并采用qRT-PCR和蛋白质印迹法鉴定其干扰效果;采用CCK-8法检测转染后MDA-MB-468细胞增殖能力;流式细胞术检测MDA-MB-468细胞转染后细胞凋亡水平;蛋白质印迹法检测MDA-MB-468细胞中PI3K/AKT信号通路以及凋亡相关蛋白cleaved-caspase-3、Bcl-2、Bax和增殖相关蛋白Ki-67等表达。结果:MDA-MB-468细胞中ACSS2 mRNA和蛋白表达水平明显高于MCF-10A细胞(P<0.01);与阴性对照组细胞相比,靶向干扰MDA-MB-468细胞ACSS2表达后细胞增殖活力明显下降(P<0.01),细胞凋亡水平明显上升(P<0.01),p-PI3K、p-AKT、Ki-67和Bcl-2表达明显降低(P均<0.01),cleaved-caspase-3、Bax表达明显上升(P均<0.01)。结论:干扰ACSS2表达可能通过PI3K/AKT信号通路抑制MDA-MB-468细胞增殖和促进凋亡发生。
Objective:To investigate the effect of acetyl-CoA synthase 2(ACSS2)on proliferation and apoptosis in triple-negative breast cancer MDA-MB-468 cell and its potential mechanism.Methods:The mRNA and protein expression of ACSS2 in normal breast epithelial cells MCF-10A and human triple-negative breast cancer MDA-MB-468 cell was detected by quantitative real-time PCR(qRT-PCR)and Western blotting,respectively.SiRNAs specific for ACSS2(siRNA-ACSS2)or a scrambled sequence(siRNA-NC)were designed and synthesized as positive and negative controls for transfection,respectively.SiRNA-ACSS2 or siRNA-NC was transiently transfected into triple-negative breast cancer MDA-MB-468 cells.The interference efficiency of ACSS2 were assessed by qRT-PCR and Western blotting.The proliferation capacity of MDA-MB-468 before and after conducting the targeted interference with ACSS2 was examined by CCK-8 assay.The apoptosis rate of MDA-MB-468 cells before and after interference was determined,using flow cytometry.Additionally,PI3K/AKT signaling pathway and the apoptosis-related proteins such as cleaved-caspase-3,Bcl-2,Bax and proliferation marker ki-67 were detected by Western blotting.Results:Compared with normal breast epithelial cells MCF-10A,MDA-MB-468 cells were showed with higher levels of the mRNA and protein expression of ACSS2(P<0.01).Compared with MDA-MB-468 cells without the interference,targeted interference of ACSS2 strongly inhibited its proliferation and remarkably induced the apoptosis in MDA-MB-468 cells(all P<0.01).The protein expression levels of p-PI3K,p-Akt,Ki-67 and Bcl-2 were significantly decreased(all P<0.01),meanwhile,the protein expression levels of cleaved-caspase-3 and Bax were marked increased(all P<0.01).Conclusion:Interference with acetyl-CoA synthase 2 dramatically inhibit the proliferation of triple-negative breast cancer MDA-MB-468 cells and significantly promote the apoptosis of breast cancer cells,which may be through PI3K/AKT signaling pathway.
作者
傅聪
周月鹏
尹超云
凌锐
浦希
汤翔
陈德玉
FU Cong;ZHOU Yuepeng;YIN Chaoyun;LING Rui;PU Xi;TANG Xiang;CHEN Deyu(Clinical Center of Tumor Therapy,Affiliated Hospital of Jiangsu University,Zhenjiang Jiangsu 212001,China;Oncological Laboratory,Affiliated Hospital of Jiangsu University,Zhenjiang Jiangsu 212001,China;Department of Vascular Surgery,Affiliated Hospital of Jiangsu University,Zhenjiang Jiangsu 212001,China)
出处
《江苏大学学报（医学版）》
CAS
2021年第5期374-379,共6页
Journal of Jiangsu University：Medicine Edition
基金
国家自然科学基金面上项目(81572956)
江苏省社会发展基金资助项目(BE2017696)
江苏省医学创新团队项目(CXTDC2016009)。
关键词
三阴性乳腺癌
乙酰辅酶A合成酶2
RNA干扰
增殖
凋亡
triple-negative breast cancer
acetyl-CoA synthetase 2
RNA interference
proliferation
apoptosis
