热疗联合奥沙利铂对SW480细胞增殖和凋亡的影响

Effect of Hyperthermia Combined with Oxaliplatin on the Proliferation and Apoptosis of SW480 Cells

目的:观察奥沙利铂联合热疗对人结肠癌细胞SW480增殖及凋亡的影响,确定联合用药的效果,为临床方案提供参考。方法:采用MTT(四唑盐)法检测热疗、奥沙利铂及联合用药对细胞增殖的影响;瑞士吉姆萨染色法观察细胞形态;流式细胞仪检测细胞凋亡和周期;Western blot检测Bax、Bcl-2以及Caspase8蛋白表达量变化;q PCR检测Bax、Bcl-2以及Caspase8 m RNA的积累。结果:热疗联合奥沙利铂可以显著抑制细胞增殖,与对照组相比,热疗组、化疗组、联合组细胞凋亡率分别为16.2%、20.5%和36.1%,具有显著性差异(P<0.01);细胞学形态中,热疗组细胞发生皱缩,化疗组细胞膜破裂;化疗将细胞阻滞在G2/M期,热疗和联合组将细胞阻滞S期;Western blot和qPCR显示Bax/Bcl-2比值上升,Caspase8表达量增加,联合组三种蛋白的表达量均与对照组具有显著性差异(P<0.01)。结论:热疗联合奥沙利铂可以显著促进细胞凋亡,提高治疗效果,为结肠癌的治疗提供参考。

Objective: To observe the effect of oxaliplatin combined with hyperthermia on the proliferation and apoptosis of human colon cancer cell line SW480, to determine the effectiveness of the combination therapy and to provide regimen for clinical reference. Methods: MTT assay was used to examine the cell proliferation after treatment with hyperthermia and/or oxaliplatin, Giemsa staining for cell morphology, and flow cytometry for cell apoptosis and cycle. The changes of Bax, Bcl-2 and Caspase8 protein were analyzed by Western blot. The expressions of Bax, Bcl-2 and Caspase8 were detected by qPCR. Results: Hyperthermia combined with oxaliplatin could significantly inhibit cell proliferation. Compared with the control group, the apoptotic rates of hyperthermia group, chemotherapy group and combination group were 16.2%, 20.5% and 36.1%, respectively, with significant increase(P<0.01);In cytological morphology,cells in hyperthermia group shrank and cell membrane rupture in chemotherapy group;The cell cycles of chemotherapy group was arrested at G2/M while hyperthermia group and combination group induced cell cycle at the S phase;Western blot and qPCR showed that the ratio of Bax/Bcl-2 and expression of Caspase8 increased, and the expression of the three proteins in the combined group was significantly different from the control group(P<0.01). Conclusions: The treatment of hyperthermia combined with oxaliplatin can significantly promote apoptosis and improve treatment, providing a reference for the treatment of colon cancer.