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

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 ...

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

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.