肠胃清逆转结肠癌HCT116/L-OHP细胞草酸铂耐药作用及对Pt-DNA加合物的影响

目的探讨中药复方肠胃清(黄芪、白术、党参、猪苓、薏苡仁、八月札、野葡萄藤、红藤等)对结肠癌HCT116/L-OHP细胞草酸铂耐药的逆转作用及对Pt-DNA加合物的影响。方法将HCT116/L-OHP细胞种植于裸小鼠皮下,建立肿瘤耐药模型,随机分为5组,A组为对照组,B组为草酸铂组,C组为肠胃清组,D组为草酸铂+肠胃清低剂量组,E组为草酸铂+肠胃清高剂量组,治疗前后测量移植瘤体积及荷瘤鼠体质量;采用细胞增殖抑制实验MTT法观察肠胃清药物血清对HCT116/L-OHP细胞株耐药性的逆转作用,并采用原子吸收分光光度法检测细胞内铂(Platinum,Pt)药物及核内Pt-DNA水平。结果肠胃清联合草酸铂组与肠胃清或草酸铂单独用药组相比,有更高的肿瘤抑制率(P0.01),其中肠胃清高剂量联合草酸铂组抑瘤率最高;与对照组相比,含肠胃清组荷瘤鼠体质量减轻好于单用草酸铂;MTT结果显示7.5%肠胃清药物血清能够增加耐药细胞株HCT116/L-OHP对草酸铂敏感性,逆转指数为2.74;肠胃清药物血清可以增加HCT116/L-OHP细胞内草酸铂水平,并能使细胞核内Pt-DNA加合物水平增加15.74%。结论肠胃清能够逆转HCT116/L-OHP细胞对草酸铂的耐药性,其机制与肠胃清增加细胞内Pt-DNA加合物水平有关。

Effects of Changweiqing in reversing oxaliplatin resistant HCT116/L-OHP and influence on Pt-DNA adduct

AIM To investigate the effects of Changweiqing(Astragali Radix,Atractylodis macrocephalae Rhizoma,Codonopsis Radix,Polyporus,Coicis Semen,Akebiae Fructus,Vitis quinquangularis Caulis,Sargentodoxae Caulis,etc.) on reversing oxaliplatin resistance in human colon cancer cell line HCT116/L-OHP and influence in Pt-DNA.METHODS The oxaliplatin resistant human colon cancer cell line nude mice xenograft model was established by inoculating HCT116/L-OHP.Then the mice were randomly divided into five groups: control group,oxaliplatin group,Changweiqing group,low dose of Changweiqing plus oxaliplatin group and high dose of Changweiqing plus oxaliplatin group.The tumor volume and mice weight in each group were calculated before and after the treatment.The effects of Changweiqing drug serum on reversing oxaliplatin-resistant HCT116/L-OHP were measured by MTT assay.The cellular accumulation of platinum and Pt-DNA adduct was determined by atomic absorption spectrophotometry.RESULTS The tumor volume in low and high dose of Changweiqing plus oxaliplatin groups was smaller than that in Changweiqing or oxaliplatin groups(P<0.01).The high dose of Changweiqing plus oxaliplatin group had the highest tumor inhibitory rate.The decrease of weight in Changweiqing plus oxaliplatin group was less than that in oxaliplatin group.The reversal index of 7.5% Changweiqing drug serum on HCT116/L-OHP reached 2.74.After being treated by 7.5% drug serum,the intracellular platinum drug and Pt-DNA adduct contents of HCT116/L-OHP cells increased,and Pt-DNA adduct contents increased by 15.74%.CONCLUSION The reversal of oxaliplatin-resistant HCT116/L-OHP by Changweiqing may be associated with its increase in Pt-DNA adduct concentration in tumor cell.