CDK9抑制剂F200对乳腺癌细胞MCF7凋亡的影响

目的研究细胞周期依赖性激酶（cell cycle dependent kinase,CDK）CDK9抑制剂F200对乳腺癌细胞MCF7凋亡的影响。方法 MCF7细胞培养于含0.01 mg·mL^-1人重组胰岛素及10%胎牛血清的RPMI 1640培养液中,待对数生长期时接种细胞进行实验,24 h贴壁后给药：分为阴性对照组（0.5%DMSO）、阳性对照组（R-Roscovitine 5.66μM）及药物组（F200 0.1μM,0.71μM）,给药48 h后利用TUNEL法染色观察细胞凋亡DNA断裂情况及细胞凋亡形态学变化、流式细胞技术检测细胞的凋亡比率、免疫印迹法检测凋亡标志蛋白PARP的表达情况。结果 TUNEL结果显示,随着F200浓度增加,细胞出现明显的固缩变圆,细胞核可见深染致密颗粒,细胞核质分界明显等凋亡表现;流式细胞仪结果显示,0.71μM的F200能够诱导32.6%的细胞凋亡;Western Blot结果显示,随着F200浓度增加,PARP蛋白剪切水平明显增加。结论 F200能有效促进乳腺癌细胞MCF7的凋亡。

Effect of CDK9 Inhibitor F200 on the Apoptosis of Human Breast Cancer MCF7 Cells

Objective This study aimed to investigate the effects of cell cycle dependent kinase inhibitor F200 on apoptosis of human breast cancer cell MCF7.Methods MCF7 cells were cultured in RPMI-1640 medium with 0.01 mg·mL^-1human recombinant insulin and 10% fetal bovine serum.Cells were subcultured at exponential growth phase.24 h later,cells were adherent to the plate and treatments were given according to groups setup,negative control（0.5% DMSO）,positive control（R-Roscovitine 5.66μM）,F200 groups（F200 0.1μM,0.71μM）.At 48 h after treatment,cell morphological change of apoptosis was measured by TUNEL methods and the apoptosis rate was detected by flow cytometry.The expression of PRAP was measured by Western Bolt.Results The TUNEL results showed that as the F200 concentration increased,the cell apoptosis features like cell pyknosis and dense granule in nucleus were significantly clearer.More cells were stained with DAB after treated with F200.The flow cytometry results showed 0.71μM F200 could induce 32.6% cell apoptosis.Moreover,the Western Blot results showed cleaved PARP was increased along with the increase of F200 concentration.Conclusion CDK inhibitor F200 could inhibit MCF7 cell growth and induce the cell apoptosis.