舒林酸硫化物对人乳腺癌细胞SKBR-3增殖与凋亡的影响

Effects of sulindac sulfide on proliferation and apoptosis of SKBR-3 cells

目的研究舒林酸硫化物对人乳腺癌细胞SKBR-3增殖与凋亡的影响及其可能机制。方法用20,40,80μmol·L-1舒林酸硫化物分别处理SKBR-3细胞24,48,72 h,正常组和对照组分别用培养基和0.1%二甲基亚砜处理相同的时间。用噻唑蓝(MTT)法和流式细胞术来观察舒林酸硫化物对SKBR-3细胞增殖和凋亡的影响,用Western blot检测细胞中Bcl-2、Bax、细胞色素C、Caspase-3、Cleaved Caspase-3蛋白水平。结果舒林酸硫化物可明显抑制SKBR-3细胞的增殖,同时促进细胞的凋亡。舒林酸硫化物在作用SKBR-3细胞24 h后,与正常组比较Bcl-2和线粒体细胞色素C蛋白水平显著下降(P<0.05),Bax、Cleaved Caspase-3和胞质细胞色素C蛋白水平显著升高(P<0.05),而Caspase-3蛋白水平差异无统计学意义(P>0.05)。结论舒林酸硫化物能够抑制细胞的增殖,促进细胞的凋亡,其促进细胞凋亡的机制可能是通过下调Bcl-2蛋白的表达与上调Bax蛋白的表达,诱导细胞色素C从线粒体中释放,最终激活Caspase-3蛋白,进而促进SKBR-3细胞的凋亡。

Objective To investigate the effects of sulindac sulfide on proliferation and apoptosis of SKBR- 3 cells,and then explore its potential molecular mechanism. Methods SKBR- 3 cells were incubated with different concentrations of sulindac sulfide（ 20,40,80 μmol·L- 1）for 24,48,72 h,normal group and control group were respectively treated with medium and 0. 1% DMSO for the same time. Apoptosis and proliferation of SKBR- 3 cells were analyzed by using flow cytometry and MTT assay,respectively. Western blot was used to test the protein expression of Bcl- 2,Bax,cytochrome C,Caspase- 3 and Cleaved Caspase- 3. Results Sulindac sulfide inhibited the proliferation of SKBR- 3 cells and promoted the apoptosis of cells. After 24 hours of incubation,the protein expression of Bcl- 2 and mitochondrial cytochrome C markedly reduced（ P〈0. 05）,the protein expression of Bax,cytosolic cytochrome C and Cleaved Caspase- 3 significantly increased（ P〈0. 05）,the protein expression of Caspase- 3 had no significant difference（ P〉0. 05）. Conclusion Sulindac sulfide significantly inhibit the proliferation of SKBR- 3 cells and promote cells apoptosis.The potential molecular mechanism of pro- apoptosis might be that sulindac sulfide down- regulates the protein expression of Bcl- 2,up- regulates the protein expression of Bax,induces the release of cytochrome c from mitochondria,finally activates the Caspase- 3 protein and then promotes the apoptosis of SKBR- 3 cells.