LukS-PV对乳腺癌细胞MAD-MB-231增殖、凋亡及细胞周期的影响

Effect of LukS-PV on proliferation, apoptosis and cell cycle of breast cancer cell line MAD-MB-231

目的在体外研究金黄色葡萄球菌杀白细胞素S组分(LukS-PV)对三阴乳腺癌(TNBC)细胞(MAD-MB-231)形态、增殖、周期和凋亡的影响。方法 0.25、0.5、0.75、1μmol/L LukS-PV刺激乳腺癌细胞MAD-MB-231后,利用倒置显微镜观察细胞形态的改变;用CCK-8检测LukS-PV对MAD-MB-231细胞增殖的影响;通过PI单染流式细胞术检测LukS-PV对MAD-MB-231细胞周期的改变;采用Annexin V/PI双染流式细胞术检测LukS-PV对MAD-MB-231细胞凋亡的影响,利用Western blot检测Bcl-2相关的X蛋白(Bax)、B淋巴细胞瘤-2 (Bcl-2)、天冬氨酸特异性半胱氨酸蛋白酶-3(Caspase3)、Caspase8表达的变化。结果 LukS-PV作用MAD-MB-231后,细胞逐渐变圆,随着浓度增加,悬浮细胞增多。LukS-PV以时间和浓度依赖性的方式抑制乳腺癌细胞的增殖。LukS-PV可使MAD-MB-231周期阻滞在G1/G0期,呈浓度依赖性。LukS-PV可诱导MAD-MB-231凋亡,上调Bax、活化的caspase3(Cleved-caspase3)、活化的caspase8(Cleved-caspase8)的表达,下调Bcl-2的表达。结论 LukS-PV可抑制MAD-MB-231增殖,使其周期阻滞在G1/G0期,并诱导其凋亡。

Objective To study the effects of S component of panton-valentine leucocidin (LukS-PV) on morphology,proliferation,cell cycle and apoptosis of triple-negative breast cancer cell line MAD-MB-231 in vitro . Methods Cells were exposed to LukS-PV at different concentrations,the cell morphology was observed under an inverted microscope. CCK-8 was used to detect the effects of LukS-PV on cell proliferation. Cell cycle was detected by flow cytometry following PI staining. Cell apoptosis was detected by flow cytometry following Annexin V/PI staining and apoptosis-related proteins were detected by Western blot. Results With different concentration of LukS-PV was applied to MAD-MB-231,the cells gradually became round and the suspended cells increased. LukS-PV inhibited the proliferation of breast cancer cells in a time- and concentration-dependent manner. LukS-PV could block MAD-MB-231 in G1/G0 phase in a concentration-dependent manner. LukS-PV could induce apoptosis of MAD-MB-231,up-regulate the expression of Bax,Cleved-caspase3 and Cleved-caspase8,and down-regulate the expression of Bcl-2. Conclusion LukS-PV can inhibit the proliferation of MAD-MB-231,arrest its cycle in G1/G0 phase,and induce its apoptosis.