17-烯丙胺-17-脱甲氧格尔德霉素对乳腺癌细胞增殖及侵袭作用

Regulatory effect of 17-AAG on the proliferation and invasion of breast carcinoma cells via STAT3 signaling pathway

目的:观察17-烯丙胺-17-脱甲氧格尔德霉素(17-allylamino-17-desmethoxy-geldanamycin,17-AAG)通过抑制转录活化蛋白3(signal transducer and activator of transcription3,STAT3)信号通路发挥对人乳腺癌MCF-7细胞增殖和侵袭的调控作用。方法:体外培养人乳腺癌MCF-7细胞,分别给予不同剂量及不同作用时间的17-AAG进行作用,MTT法检测细胞增殖能力;RT-PCR法和Western Blot法检测各组细胞STAT3、血管内皮生长因子(vascular endothelial growth factor,VEGF)、基质金属蛋白酶2(matrix metalloproteinase2,MMP-2)和MMP-9 mRNA和蛋白的表达水平,Transwell小室检测细胞侵袭能力。结果:0.165～10mg/L的17-AAG作用24 h、48 h后对MCF-7细胞有显著的抑制作用;1.0、2.0、3.0、5.0 mg/L的17-AAG作用48 h后,各组细胞STAT3、VEGF、MMP-2和MMP-9 mRNA和蛋白表达均下调,且具有浓度依赖性;Tran-swell小室检测显示17-AAG处理后穿膜细胞数明显减少,与空白对照组相比差异均有统计学意义(P<0.05或P<0.01)。结论:17-AAG对人乳腺癌MCF-7细胞的增殖具有明显的抑制作用,并能够抑制STAT3的mRNA和蛋白的表达水平,进而抑制VEGF、MMP-2和MMP-9 mRNA及蛋白的表达水平,从而调控乳腺癌细胞的侵袭性。

Objective： To investigate the effects of 17 - AAG on the proliferation and invasion of human breast carcinoma MCF -7 cells via STAT3 signaling pathway. Methods： MCF -7 cells were cultured in DMEM medium, MTT method was performed to evaluate the inhibitory effects of 17 - AAG with different time and different dose on proliferation of MCF - 7 cells, the genes and proteins expressions of STAT3, VEGF, MMP - 2 and MMP - 9 were detected by RT - PCR and Western Blotting analysis, invasion and migration of MCF - 7 were investigated with transwell chamber test. Results ： MCF - 7 cells were treated with 0. 165 - 10 mg/L 17 - AAG for 24 and 48 hours and the proliferation was significantly inhibited by 17 - AAG. The assay of RT - PCR and Western Blotting showed that the genes and proteins expressions of STAT3, VEGF, MMP - 2 and MMP - 9 in MCF - 7 cells induced by 17 - AAG were significantly down - regulated in a dose - dependent manner when the cells were treated with 1.0, 2.0, 3.0, 5.0 mg/L 17 - AAG for 48 hours, which was supported by the results of transwell chamber test. Conclusion： 17 - AAG can inhibit the cell proliferation and migration by down - regulating the genes and proteins expressions of STAT3, VEGF, MMP -- 2 and MMP - 9 in human breast carcinoma MCF -7 cells in vitro.