褪黑素联合顺铂通过诱导细胞凋亡抑制人胶质瘤细胞U251和SHG-44增殖

Melatonin combined with cisplatin suppresses proliferation of human glioma cells U251 and SHG-44 by inducing cell apoptosis

目的:研究褪黑素(melatonin,MT)和顺铂(cisplatin,DDP)单独或联合应用对人胶质瘤细胞U251和SHG-44增殖及凋亡的影响。方法:采用不同质量浓度MT和DDP单独或联合处理U251和SHG-44细胞,并设对照组(不加任何药物)及乙醇组(加入乙醇);以CCK-8法检测细胞的增殖,流式细胞术检测细胞的凋亡和细胞周期,采用两药相互作用指数(coefficient of drug interaction,CDI)评估MT是否影响U251和SHG-44细胞对DDP的敏感性。结果:CCK-8结果显示,单用MT或DDP可浓度依赖性抑制U251和SHG-44细胞的增殖,MT还可协同增强DDP对U251和SHG-44细胞的增殖抑制作用(CDI<1)。流式细胞术检测结果显示,MT可促进U251和SHG-44细胞的凋亡,MT可增强DDP对U251和SHG-44细胞的凋亡诱导作用,0.5 mmol/L MT联合20μg/ml DDP组U251和SHG-44的细胞凋亡率显著高于20μg/ml DDP组[(66.3±1.0)%vs(45.9±1.7)%,(35.5±0.8)%vs(15.5±0.8%);均P<0.01];而且0.5 mmol/L MT联合20μg/ml DDP组U251和SHG-44细胞的G1期比例显著高于20μg/ml DDP组[(52.4±2.1)%vs(27.9±1.5)%,(39.7±1.5)%vs(27.7±1.3)%;均P<0.01]。结论:MT能显著增强DDP对人胶质瘤细胞U251和SHG-44的凋亡诱导作用,从而协同增强DDP对细胞增殖的抑制作用,有望成为人胶质瘤化疗的辅助药物。

Objective ： To investigate the effect of melatonin （MT） and cisplatin （DDP） used alone or in combination on proliferation and apoptosis of human glioma cells U251 and SHG-44. Methods： U251 and SHG-44 cells were treated with various concentrations of MT and DDP alone or in combination, and the control group （without adding any drug） and ethanol group （adding the ethanol） were included. CCK-8 assay was used to detect cell proliferation, and flow cytometry was used to detect cell apoptosis and cell cycle. The coefficient of drug interaction （CDI） was used to evaluate whether MT could affect the sensitivity of U251 and SHG-44 cells to DDP. Results： CCK-8 results showed that MT or DDP used alone inhibited the proliferation of U251 and SHG-44 cells in a concentration dependent manner, and 0.5 mmol/L MT synergistically enhanced the proliferation inhibitory effect of DDP on U251 and SHG-44 cells （CDI〈1）. Flow cytometry results showed that MT promoted U251 and SHG-44 cell apoptosis, and MT enhanced the apoptosis inducing effect of DDP on U251 and SHG-44 cells. The apoptotic rate of U251 and SHG-44 cells in the 0.5 mmol/L MT and 20 μg/ml DDP combination group was significantly higher than that in 20 μg/ml DDP group （／[66.3±1.0／]% vs ／[45.9±1.7／]%, ／[355±0.8／]% vs ／[15.5±08／]%, P〈0.01）. And the percentage of U251 and SHG-44 cells in sub-G1 phase in the 0.5 mmol/L MT and 20 μg/ml DDP combination group was significantly higher than that in 20 μg/ml DDP group （／[524±2.1／]% vs ／[27.9±1.5／]%, ／[39.7±1.5／]% vs ／[27.7±1.3／]%, P〈0.01／]. Conclusion： MT can enhance the apoptosis inducing effect of DDP on human glioma cells U251 and SHG-44, thereby synergistically enhancing the suppressive effects of DDP on human glioma cell proliferation, which may be used as a complementary drug in human glioma chemotherapy.