尼卡地平对结肠癌细胞HCT116/L耐药的逆转作用研究

The Reversal Effect of Nicardipine on HCT116/L Colon Cancer Cells

目的探究尼卡地平逆转耐奥沙利铂结肠癌细胞HCT116/L耐药的疗效及其机制。方法采用MTT法分别检测尼卡地平和奥沙利铂在HCT116/L和HCT116细胞上的药敏性,并检测尼卡地平能否逆转奥沙利铂耐药。采用流式细胞仪(FCM)检测HCT116/L细胞凋亡和细胞内罗丹明-123的浓度。结果奥沙利铂在耐药细胞HCT116/L及亲本细胞HCT116的IC50分别为(20.07±1.32)μM,(0.65±0.05)μM,具有明显的耐药差异性(P<0.001),耐药指数(RI)为(30.86±2.21)。尼卡地平在耐药细胞HCT116/L及亲本细胞HCT116的IC50分别为(22.76±1.52)μM,(21.65±1.35)μM,两者之间的细胞毒性没有统计学差异。FCM法检测结果显示尼卡地平和奥沙利铂联用能显著增加耐药细胞株凋亡率(P<0.001),且能明显增加耐药细胞HCT116/L内罗丹明-123的浓度。结论尼卡地平能增强HCT116/L细胞对奥沙利铂的敏感性,其逆转耐药的作用与诱导耐药细胞HCT116/L凋亡,影响转运蛋白P-gp的功能有关。

OBJECTIVE To study the mechanism of nicardipine reverse cell resistance in HCT116/L cells,and to detect the effect of combination of nicardipine and oxaliplatin on HCT116/L cells.METHODS MTT method was used to detect the drug sensitivity of nicardipine and oxaliplatin on HCT116/L and HCT116 cells,and to detect whether nicardipine could reverse oxaliplatin resistance.Flow cytometry(FCM)was used to detect the apoptosis of HCT116/L cells and the content of Rho123 in HCT116/L cells.RESULTS The IC50 of oxaliplatin in drug-resistant cells HCT116/L and parental cells HCT116 were(20.07±1.32)μM and(0.65±0.05)μM,respectively,showing significant differences in drug resistance(P<0.001).And the resistance index(RI)was 30.86±2.21.The IC50 of nicardipine in HCT116/L and HCT116 were(22.76±1.52)μM and(21.65±1.35)μM,respectively.There was no statistical difference in cytotoxicity between the two cells.FCM assay showed that nicardipine combining with oxaliplatin could significantly increase the apoptosis rate of HCT116/L(P<0.001),and significantly increase the content of Rho123 in drug-resistant cells HCT116/L.CONCLUSION Nicardipine can reverse the resistance of HCT116/L cells to oxaliplatin.The effect of reversing resistance are related to induce the apoptosis of HCT116/L cells and affect the function of the P-gp transport protein.