温热刺激增强卵巢癌细胞对奥沙利铂(oxaliplatin) 化疗的敏感性

Thermotherapy Enhanced the Sensitivity of Ovarian Cancer Cell to Oxaliplatin

目的探讨温热刺激对卵巢癌细胞化疗效果的影响与潜在作用机制.方法首先采用Cell Counting Kit-8(CCK8)细胞毒性实验检测温热刺激下,奥沙利铂对卵巢癌细胞A2780作用半数致死浓度(IC50)的变化;然后采用流式细胞术检测温热刺激对卵巢癌细胞凋亡的影响.最后采用电感耦合等离子体-质谱法(inductively coupled plasma-mass spectrometry,ICP-MS)检测细胞内铂浓度,并经qPCR检测细胞膜铜转运蛋白1(CTR1)与有机阳离子转运蛋白(OCT1-3)mRNA表达情况.结果与单纯药物处理组相比,温热刺激能显著降低奥沙利铂对卵巢癌细胞的IC50(t=3.47,P=0.0084),还能显著增加奥沙利铂诱导卵巢癌细胞凋亡现象的发生(t=10.42,P=0.0091).与单纯药物处理组相比,细胞摄入的铂含量在热刺激后增加(t=3.198,P=0.0494),CTR1(t=19.79,P<0.0001)与OCT1-3(OCT1:t=16.02,P=0.005;OCT3:t=7.95,P=0.0169)转录水平表达量也显著增加.结论温热刺激能增强卵巢癌细胞A2780的化疗效果,作用机制可能与其CTR1与OCT1-3的表达、促进细胞对铂类药物的摄入有关.

Objective To explore the effects and mechanisms of thermotherapy on the Oxaliplatin treated ovarian cancer cells.Methods Firstly,Cell Counting Kit-8(CCK8)cytotoxicity assay was used to detect the effects of thermotherapy on the half maximal inhibitory concentration(IC50)of Oxaliplatin for ovarian cancer cell line A2780.Next,flow cytometry analysis was performed to identify the effects of thermotherapy on Oxaliplatininduced apoptosis of A2780 cells.Finally,ICP-MS was adopted to determine the intracellular platinum concentration in response to thermotherapy and/or Oxaliplatin treatment,followed by qPCR mRNA expression confirmation of cell membrane copper transporter 1(CTR1)and organic cation transporters 1-3(OCT 1-3).Results Compared with the Oxaliplatin treatment group,thermotherapy could significantly reduce the IC50 of Oxaliplatin in ovarian cancer cells(t=3.47,P=0.0084),and enhance the Oxaliplatin-induced apoptosis of ovarian cancer cells(t=10.42,P=0.0091).Compared with Oxaliplatin treatment group,the intracellular platinum was significantly increased under thermotherapy(t=3.198,P=0.0494),which was consistent with the variation of mRNA expression pattern for CTR1(t=19.79,P<0.0001)and OCT 1-3(OCT1:t=16.02,0.005;OCT3:t=7.95,P=0.0169)after heat stimulation.Conclusion Thermotherapy could enhance the effect of chemotherapy on human ovarian cancer cells by enhancing Oxaliplatin-induced cell apoptosis.The underlying mechanism was related to the thermotherapy-induced increase of CTR1 and OCT 1-3 mRNA expression,through which facilitating intracellular Oxaliplatin uptake.