抑制Cks1表达对乳腺癌MCF-7细胞增殖、侵袭转移能力的影响及机制探讨

Effect and mechanism of Cks1 RNAi on the proliferation,migration and invasion of breast cancer cells

目的观察抑制Cks1表达对人乳腺癌MCF-7细胞增殖、侵袭转移能力的影响,并探讨其机制。方法将MCF-7细胞分为两组,观察组细胞经脂质体转染Cks1 siRNA,对照组细胞转染Scrambled siRNA;分别采用MTS法、Transwell小室实验观察MCF-7细胞的增殖及侵袭转移能力,采用Western blot法检测MCF-7细胞中的MMP-2、MMP-9蛋白。结果观察组转染608、4、1081、32 h后,MCF-7细胞的OD值分别为0.15±0.01、0.3±0.02、0.7±0.01、1.3±0.02,对照组分别为0.2±0.010、.5±0.01、1.3±0.03、1.8±0.02,两组转染84、108、132 h细胞OD值比较,P均<0.05。观察组转染48 h,侵袭细胞数为(150±17)个、转移细胞数为(210±22)个,对照组分别为(550±90)(、660±96)个,两组比较,P均<0.05。转染后12 h,观察组MMP-2、MMP-9蛋白表达量为0.05±0.0020、.10±0.002,对照组分别为0.95±0.020、0.98±0.030,两组比较,P均<0.05。结论抑制Cks1表达可显著抑制MCF-7细胞的增殖、侵袭转移能力,该作用可能与MMP-2、MMP-9蛋白表达降低有关。

Objective To investigate the effect of Cksl invasion of breast cancer cells, and explore the mechanism. RNA interference （RNAi） on the proliferation, migration and Methods The method of RNAi was used to knock down the expression of Cksl and Western-blot was used to detect the MMP-2,MMP-9. MTS and Transwell experiment was performed to examine the proliferation, migration and invasion ability of MCF-7 cells, respectively. Results 60, 84, 108, 132 h af- ter transfection, the OD value was 0.15 ± 0. 01,0.3 ± 0. 02,0.7 ± 0. 01,1.3 ± 0. 02 in Cksl knock down group, respectively. But in control group the OD value was 0. 2 ± 0. 01,0.5 ± 0. 01,1.3 ± 0. 03,1.8 ± 0. 02, respectively： Beside 24 h, significant different were found between corresponding time point in these two groups （P 〈 0.05）. The number of invasion and migration ceils was 150 ± 17 and 210 ±22, in Cksl knock down group. But in control group, it was 550 ±90 and 660 ± 96, respectively. Significant different was found between two groups （all P 〈 0. 05）. Moreover, the expression of MMP- 2（0.05 ±0.002）, MMP-9（0.10 ± 0.002） sifnificantly decreased after Cksl knockdown （ all P 〈 0.05 ）. Conclusion Cksl expression has effects on the migration and invasion ability of MCF-7 cells and this effect is accomplished at least partly through MMP-2 and MMP-9.