干扰乙酰辅酶A合成酶2对三阴性乳腺癌MDA-MB-468细胞增殖及凋亡的影响

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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MB468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。

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	傅聪1
	周月鹏2
	尹超云3
	凌锐1
	浦希1
	汤翔1
	陈德玉1

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

Abstract：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.

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收稿日期: 2020-12-15

基金资助: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）

通讯作者: 陈德玉（通讯作者），教授，博士生导师，E-mail: cdeyu@hotmail.com

作者简介: 傅聪（1995— ），女，硕士研究生

引用本文:

傅聪1，周月鹏2，尹超云3，凌锐1，浦希1，汤翔1，陈德玉1. 干扰乙酰辅酶A合成酶2对三阴性乳腺癌MDA-MB-468细胞增殖及凋亡的影响[J]. 江苏大学学报:医学版, 2021, 31(05): 374-379. FU Cong1， ZHOU Yuepeng2， YIN Chaoyun3， LING Rui1， PU Xi1， TANG Xiang1， CHEN Deyu1.

#br# Effect of acetyl-CoA synthase 2 interference on the proliferation and apoptosis of human triple-negative breast cancer MDA-MB-468 cell lines

. Journal of Jiangsu University(Medicine Edition), 2021, 31(05): 374-379.

链接本文:

http://zzs.ujs.edu.cn/xbyxb/CN/ 或 http://zzs.ujs.edu.cn/xbyxb/CN/Y2021/V31/I05/374

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