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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 study the effect of Chinese herbal compound “Changwei Wei” (CWQ) combined with oxaliplatin (L-OHP) on invasiveness of human colon carcinoma oxaliplatin-resistant cell line HCT116 / L-OHP in vitro. Methods HCT116 and HCT116 / L-OHP cells were cultured in vitro and the cell proliferation was evaluated by MTT assay. The drug resistance index (RF) was calculated by IC50. The reversal index of RI ). Transwell experiments were used to observe the change of invasion ability of drug-resistant cells. The rats were divided into 5 groups: control group, 2 doses of low-dose oxaliplatin group and 2 doses of oxaliplatin low-dose combined with gastrointestinal extract group. The cells were divided into 4 groups: control group, oxaliplatin group, Changwei group and Weishangqing combined oxaliplatin group. The cells in each group were treated for 48 hours. Results The stability of HCT116 / L-OHP resistant human colon cancer cell line was good, with a RF of 10.49. Gastrointestinal combined with oxaliplatin significantly inhibited the proliferation of HCT116 / L-OHP-resistant cells, and the reversal index (RI) was 4.46. The inhibition rate of oxaliplatin high-dose combined with Gutta-extract (48.34%) was significantly higher than that of high-dose oxaliplatin (19.83%) (P <0.01). The gray value of ADAM-17, MMP-2 and MMP-9 in combination group were significantly lower than that in oxaliplatin group (1.14,0.96,0.26, respectively), the gray values were 0.53,0.68,0.05 Statistically significant (P <0.05). CONCLUSION: Gastrointestinal decoction combined with oxaliplatin can significantly inhibit the proliferation and invasion of HCT116 / L-OHP-resistant cells, and its inhibitory effect is related to the down-regulation of ADAM-17, MMP-2 and MMP-9 proteins.