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目的:研究辛二酰苯胺异羟肟酸(suberoylanilide hydroxamic acid,SAHA)对结肠癌细胞系HCT116和SW480增殖、周期和凋亡的影响,并对其分子作用机制进行初步探讨。方法:将不同浓度SAHA分别处理结肠癌HCT116和SW480细胞后,MTT法检测SAHA对HCT116和SW480细胞增殖的影响,流式细胞仪检测HCT116和SW480细胞周期和细胞凋亡率,罗丹明(rhodamine)123和二氯二氢荧光素二乙酸酯(DCFH-DA)法检测HCT116和SW480细胞线粒体跨膜电位(ΔΨm)和活性氧(ROS)水平,Real-time PCR和Western blotting法检测乙酰化组蛋白3(Ac-H3)、p21、Cyclin D1、Bax和Bcl-2的mRNA和蛋白的表达水平。结果:SAHA作用于HCT116和SW480细胞48 h后,细胞增殖被抑制、细胞周期G1期比率升高、凋亡率升高(均P<0.05),线粒体跨膜电位显著下降、细胞内ROS产生增多(均P<0.05)。与对照组比较,SAHA处理组p21和Bax mRNA增多、Cyclin D1和Bcl-2 mRNA表达量减少(均P<0.05),相关蛋白Ac-H3、p21和Bax增多,Cyclin D1和Bcl-2减少(均P<0.05)。结论:SAHA抑制结肠癌HCT116和SW480细胞增殖、阻滞细胞周期并诱导细胞凋亡,其机可能与调节p21、Cyclin D1和Bcl-2家族基因的表达、促进组蛋白乙酰化有关。
AIM: To investigate the effects of suberoylanilide hydroxamic acid (SAHA) on the proliferation, cell cycle and apoptosis of human colon cancer cell lines HCT116 and SW480 and to investigate the molecular mechanisms. Methods: The effects of SAHA on the proliferation of HCT116 and SW480 cells were detected by MTT assay. The cell cycle and apoptosis rate of HCT116 and SW480 cells were detected by flow cytometry. The expressions of rhodamine, The mitochondrial transmembrane potential (ΔΨm) and reactive oxygen species (ROS) in HCT116 and SW480 cells were detected by flow cytometry (FCM), flow cytometry (FCM) and flow cytometry (FCM) Protein 3 (Ac-H3), p21, Cyclin D1, Bax and Bcl-2 mRNA and protein expression levels. Results: SAHA inhibited the proliferation of HCT116 and SW480 cells for 48 h, the ratio of cell cycle G1 phase increased and the apoptosis rate increased (all P <0.05), the mitochondrial transmembrane potential decreased significantly and the intracellular ROS production increased (All P <0.05). Compared with the control group, the expression of p21 and Bax mRNA increased, the expression of Cyclin D1 and Bcl-2 mRNA decreased (all P <0.05) and the levels of Ac-H3, p21 and Bax, Cyclin D1 and Bcl- All P <0.05). Conclusion: SAHA can inhibit the proliferation of colon cancer HCT116 and SW480 cells, arrest the cell cycle and induce the apoptosis. The mechanism may be related to regulating the expression of p21, Cyclin D1 and Bcl-2 family genes and promoting the histone acetylation.