从新陈代谢角度研究二甲双胍对多发性骨髓瘤细胞的作用及机制

摘要

Objective:To explore the mechanism of metformin combined with melphalan in human multiple myeloma cell line RPMI8226 from the metabolic point of view,and to provide foundation for metformin treatment and train of thought for changing the metabolic state of malignant tumors.Methods:The drugs were prepared according to a certain concentration gradient.Then we used a series of tests to detect the drug sensitivity,apoptotic rate,DNA damage and the concentration of ATP in treating RPMI8226 cell by melphalan combined with metformin or not.SPSS 17.0 was used to analyze the data.Results:The inhibitory effect of melphalan on RPMI8226 cells was significantly increased after metformin was added(P<0.01),and the inhibitory effect was enhanced with the increase of melphalan concentration.The comet assay showed that metformin increased melphalan-induced DNA damage and the apoptotic rate was increased from(12.7±2.08)%to(18.8±1.5)%(P<0.05).In the ATP concentration test,the concentration of ATP in the tumor cells was significantly decreased from(0.42±0.01) μmol/L to (0.08±0.02) μmol/L(P<0.05).Conclusion:Metformin can promote DNA damage induced by melphalan and decrease the concentration of ATP in the process of repairing DNA damage,hindering the anti-apoptotic process of tumor cells,and increase the sensitivity of melphalan to multiple myeloma cells.%目的:从新陈代谢的角度探究二甲双胍联合马法兰对多发性骨髓瘤RPMI8226细胞的作用机制,为二甲双胍应用于恶性肿瘤的治疗提供理论依据,为改变恶性肿瘤的代谢状态提供思路.方法:按照一定的浓度梯度配置好药物,采用CCK8法检测二甲双胍使用前后马法兰对肿瘤细胞化疗效应的变化,彗星实验检测DNA的损伤,流式细胞术检测细胞的凋亡,ATP生物发光检测实验检测各组药物处理后肿瘤细胞中ATP的浓度.用软件SPSS 17.0进行数据结果统计学分析.结果:加用二甲双胍后,马法兰对多发性骨髓瘤RPMI8226细胞的抑制率明显增加(P<0.01),随着马法兰浓度的增加,抑制作用增强.彗星实验表明二甲双胍促进马法兰诱导DNA损伤增加,细胞凋亡率由(12.7±2.08)%升高至(18.8±1.5)%(P<0.05).ATP浓度检测实验中,二甲双胍的使用可以明显减少肿瘤细胞中ATP的浓度,由(0.42±0.01) μmol/L降至(0.08±0.02) μmol/L(P<0.05).结论:二甲双胍促进马法兰诱导DNA损伤,并在DNA损伤亟需修复的过程减少ATP的浓度,阻碍肿瘤细胞抗凋亡过程,从而增加马法兰对多发性骨髓瘤细胞化疗敏感性.