PU-H71对结直肠癌HCT-15细胞凋亡和周期的影响

Effects of PU-H71 on Apoptosis and Cycle of HCT-15 Cells in Colorectal Cancer

目的探究PU-H71对结直肠癌HCT-15细胞增殖、凋亡、周期等生物学行为的影响,在分子水平探讨HCT-15细胞凋亡和周期的作用机制。方法体外培养人结直肠腺癌HCT-15细胞,MTT检测不同时间点和浓度的PU-H71对结直肠癌HCT-15细胞增殖的作用;流式细胞术检测细胞凋亡和周期;Western blot检测相关因子的表达水平。结果PU-H71作用24h和48h后,对结直肠癌HCT-15细胞有显著的抑制作用,并有明显的时间和浓度依赖性;设置不同浓度组作用24h后,HCT-15细胞凋亡率上升且处于G2期的细胞增多;STAT3和JAK2蛋白的表达水平明显下降,周期及凋亡相关因子CyclinD1和Bcl-2蛋白表达水平下降,Cytc、Caspase9、Caspase3、Bax蛋白表达水平上调。结论PU-H71对结直肠癌HCT-15细胞的增殖有明显的抑制作用,使细胞G2周期阻滞并诱导凋亡。PU-H71有可能通过负向调控JAK-STAT3途径来调控线粒体信号通路,以此影响凋亡和周期相关因子的表达。

Objective To study the effects of PU-H71 on the proliferation,apoptosis,cycle and other biological behaviors of colorectal cancer HCT-15 cells,and to explore the mechanism of apoptosis and cycle at the molecular level.Methods HCT-15 cells were cultured in vitro.The effects of PU-H71 at different time and concentration on the proliferation of HCT-15 cells were examined by MTT assay.The apoptosis and cell cycle of HCT-15 cells were examined by flow cytometry assay,western blot was used to detect the expression of apoptosis and cycle-related factors.Results After 24h and 48h treatment,PU-H71 showed significant inhibitory effect on colorectal cancer HCT-15 cells in time and concentration dependent manner.After treatment with PU-H71 at two concentrations of 50μg/L and 75μg/L without drug for 24h,the apoptosis rate of HCT-15 cells increased and the proportion of G2 phase cells increased.The protein expression levels of STAT3 and JAK2 were significantly decreased,the protein expression levels of CyclinD1 and Bcl-2 were decreased,and the protein expression levels of Cytc,Caspase9,Caspase3,and Bax were up-regulated.Conclusion PU-H71 can significantly inhibit the proliferation of colorectal cancer HCT-15 cells,arrest the cell cycle in G2 phase and induce cell apoptosis.PU-H71 may regulate the mitochondrial signaling pathway by negatively regulating the JAK-STAT3 pathway,thus affecting the expression of apoptosis and cyclic-related factors.