Effects of metformin on breast cancer cell proliferation, the AMPK pathway and the cell cycle
Effects of metformin on breast cancer cell proliferation, the AMPK pathway and the cell cycle
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DOI：
10.1007/s12094-013-1144-8
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发表时间：
2014-08-01
影响因子：
3.4
通讯作者：
Thompson, A. M.
中科院分区：
文献类型：
--
作者：
Hadad, S. M.;Hardie, D. G.;Thompson, A. M.
本研究的目的是比较二甲双胍对雌激素受体（ER）阳性和ER阴性乳腺癌细胞系的作用和作用机制，二甲双胍和腺苷单磷酸活化蛋白激酶（AMPK）的直接激活剂A-769662，对MCF-7（ER阳性）和MDA-MB-231（ER阴性）乳腺癌细胞系的作用通过MTT（3-（4，5-二甲基噻唑-2-基）-2，5-二苯基溴化四唑，一种黄色四唑）测定来评估。还使用了细胞活化细胞分选来检查二甲双胍对细胞周期的影响。最后，使用免疫印迹法检测二甲双胍靶AMPK及其潜在下游靶包括乙酰辅酶A羧化酶（ACC）、p53、p70-S6 K和Raptor的磷酸化。二甲双胍和A-769662在MCF-7和MDA-MB-231细胞中均引起显著的浓度依赖性细胞增殖抑制和G1细胞周期停滞。对MCF-7细胞的增殖抑制作用更明显。二甲双胍处理后，AMPK的磷酸化呈浓度依赖性，ACC的磷酸化也呈浓度依赖性，但p53、p70-S6 k或Raptor均未检测到。二甲双胍在体外对ER阳性和ER阴性乳腺癌细胞均起生长抑制作用，并使细胞停滞在G1期，特别是在ER阳性MCF-7细胞中。这种作用可能是由AMPK激活介导的，部分是通过ACC磷酸化抑制脂肪酸合成。
The aim of this study was to compare the effects and mechanisms of action of metformin on estrogen receptor (ER)-positive and ER-negative breast cancer cell lines.The anti-proliferative effects of metformin, and of the direct activator of adenosine monophosphate-activated protein kinase (AMPK), A-769662, on MCF-7 (ER-positive) and MDA-MB-231 (ER-negative) breast cancer cell lines were evaluated by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide, a yellow tetrazole) assays. Fluorescence-activated cell sorting was also used to examine the effect of metformin on the cell cycle. Finally, phosphorylation of the metformin target AMPK, and of its potential downstream targets including acetyl-CoA carboxylase (ACC), p53, p70-S6K and Raptor, was examined using immunoblotting.Metformin and A-769662 caused significant, concentration-dependent suppression of cell proliferation with G1 cell cycle arrest in both MCF-7 and MDA-MB-231 cells. The proliferation suppression effect was more profound in MCF-7 cells. A concentration-dependent phosphorylation of AMPK was detected following metformin treatment, as was phosphorylation of ACC in both cell lines, but not p53, p70-S6k or Raptor.Metformin acts as a growth inhibitor in both ER-positive and ER-negative breast cancer cells in vitro, and arrests cells in G1 phase, particularly in the ER-positive MCF-7 cells. The effect is likely to be mediated by AMPK activation, in part by inhibition of fatty acid synthesis via ACC phosphorylation.