肠胃清联合奥沙利铂对人结肠癌耐药细胞株HCT116/L-OHP侵袭作用的影响

Research on invasion effect of oxaliplatin combined with Changweiqing towards human colon cancer resistant cells HCT116/L-OHP

目的研究中药复方肠胃清(CWQ)与奥沙利铂(L-OHP)联用对结肠癌奥沙利铂耐药细胞HCT116/L-OHP体外侵袭能力的影响。方法体外培养人结肠癌奥沙利铂敏感细胞HCT116、耐药细胞株HCT116/L-OHP,MTT法观察细胞的增殖能力,根据IC50计算耐药指数(RF),检测肠胃清的逆转指数(RI)。用Transwell实验观察耐药细胞侵袭能力改变情况,分5组:对照组、低高2个剂量奥沙利铂组、奥沙利铂低高2个剂量联合肠胃清提取物组。以免疫印迹技术检测侵袭转移相关蛋白的表达,细胞分4组:对照组,奥沙利铂组,肠胃清组,肠胃清联合奥沙利铂组,各组均作用细胞48 h。结果人结肠癌HCT116/L-OHP耐药细胞株的稳定性良好,RF为10.49。肠胃清联合奥沙利铂可明显抑制HCT116/L-OHP耐药细胞增殖,逆转指数(RI)为4.46。奥沙利铂高剂量联合肠胃清提取物组的侵袭抑制率(48.34%)明显高于奥沙利铂高剂量组(19.83%),差异有统计学意义(P<0.01)。与奥沙利铂组的灰度值分别为1.14,0.96,0.26比较,联合组ADAM-17、MMP-2及MMP-9蛋白表达均显著下调,灰度值分别为0.53,0.68,0.05,差异具有统计学意义(P<0.05)。结论肠胃清联合奥沙利铂能显著抑制HCT116/L-OHP耐药细胞的增殖和侵袭能力,其侵袭抑制作用与ADAM-17、MMP-2、MMP-9蛋白下调有关。

Objective To investigate the invasion effect of the combina- tion of oxaliplatin （ L - OHP） with Changweiqing （ CWQ ） on drug - resis- tant human colon cancer cells HCT116/L- OHP in vitro. Methods Human colon cancer oxaliplatin sensitive cells HCT116 and resistant cells line HCTll6/L- OHP were cultured in vitro. The cell proliferation was detected by MTT. According to the formula of ICs0, the resistance index （RF） , combined with CWQ, CWQ reversal index （ RI ） were detected. The tumor cell invasiveness was observed by Transwell. The experimental objects （ HCT116/L - OHP） were divided 5 groups ： control group, low concentration L - OHP group, high concentration L - OHP group, low concentration L- OHP combined with CWQ extract group, high concen- tration L - OHP combined with CWQ extract group. The expression of in- vasion/ metastasis -related proteins were detected by Western blotting, resistant cells were divided 4 groups： control group, L - OHP group, CWQ group, L - OHP combined with CWQ group, effected for 48 hours.Results The drug resistance is stability in human colon cancer HCTll6/L -OHP cell lines, the resistance index （RF） is 10. 49. CWQ combined with L- OHP can obviously inhibit HCTll6/L- OHP drug -resistant cell prolifera- tion, resistance reversal index （RI） is 4. 46. Compared with L - OHP group （ ICs0 = 165.05 μg · mL^-1 ）, the cell proliferation ability was significantly decreased in the high concentration L - OHP combined with CWQ group （IC50 =36. 97 μg· mL^-1）, the difference was significant （P 〈 0. 01 ）. Compared with high concentration L- OHP group （19.83%）, the invasion inhibition rate was significantly higher in the combined CWQ group（48.34% ）, the difference was significant （P 〈 0. 01 ）. Compared with L - OHP group （proteins gray value were 1.14, 0. 96 and 0. 26, respectively）, the proteins expression of ADAM - 17, MMP -2 and MMP -9 were significantly lower in the combined experimental group （ proteins gray value were O. 53, 0. 68 and 0. 05, respectively） , the differences were significant （P 〈0. 05）. Conclusion Changweiqing combined with oxaliplatin could significantly inhibited proliferation and invasion ability of HCT116/L- OHP, the mechanism was associated with ADAM- 17, MMP- 2, MMP- 9 protein down - regulation.