肠胃清逆转耐草酸铂结肠癌细胞的DNA损伤修复实验研究

目的:观察肠胃清药物血清联合化疗药物干预HCT116、HCT116/L-OHP细胞DNA损伤修复情况的影响,进一步探讨肠胃清增效的分子机理。方法:采用MTT法,观察肠胃清药物血清的增效作用;采用Western blot方法,从DNA损伤修复的两条通路,观察L-OHP、肠胃清药物血清对XRCC1、XRCC2、XPF、ERCC1蛋白的影响,比色测定法检测DNA损伤指标AP位点。结果:肠胃清药物血清可逆转HCT116/L-OHP多药耐药。HCT116/L-OHP中XRCC1、XRCC2、XPF、ERCC1的含量较HCT116均显著增多(p<0.05),AP位点表达较HCT116显著减少(p<0.05),HCT116经L-OHP、L-OHP+肠胃清药物血清联合作用后XRCC1、XRCC2、ERCCl蛋白含量均显著减少(p<0.05),AP位点表达均显著增加(p<0.05),HCT116/L-OHP中L-OHP联合肠胃清药物血清组与对照组相比XRCC2、XPF、ERCC1蛋含量有减少趋势。结论:结肠癌耐药细胞胞内的AP位点降低,进而增加细胞的修复能力,提高耐药性。L-OHP攻击结肠癌细胞,是通过降低BER、NER的能力,导致DNA损伤加重,干扰DNA复制,达到治疗目的。肠胃清药物血清能增加L-OHP的该项能力。

Research on the DNA Damage and Repair of Reversibility of Chang Wei Qing on Drug Resistance in Oxaliplatin-Resistant Human Colon Carcinoma Cell Line

Objective:According to traditional Chinese medicine theory,the reversal effect and the molecule mechanism of Changweiqing(CWQ) on multidrug resistance(MDR) in human colorectal cancer cell were investigated,which is based on principle of fortifying spleen and detoxifying.Methods:Using MTT techniques,observation on HCT116 and HCT116/L-OHP growth inhibition for Changweiqing serum synergies;For the base excision repair proteinum:XRCC1,XRCC2,and nucleotide excision repair proteinum :ERCC1,XPF,using Western blot method,observed the effect on L-OHP,Changweiqing serum by the above DNA damage repair pathways;As AP site’ s level for DNA damage,observed L-OHP,Changweiqing serum on the level of AP sites.Results:Changweiqing scrum can reverse HCT116/L-OHP’ s multi-drug resistant.XRCC1,XRCC2,XPF,ERCC1 levels in HCT116/L-OHP are significantly increased compared with HCT116(P<0.05),AP sites significantly reduced the expression of HCT116(P<0.05),HCT116 by L-OHP, L-OHP + Changweiqing serum combined effects,XRCC1,XRCC2,ERCC1 protein were significantly decreased(P<0.05),AP sites were all significantly increased(P<0.05),HCT116/L-OHP in the L-OHP combined Changweiqing serum group compared with control group XRCC2,XPF,ERCC1 protein content is decreasing.Conclusiomn:hangweiqing can reverse MDR of HCT116/L-OHP cells,increasing the inhibition of L-OHP on HCT116,HCT116/L-OHP,mechanism is reducing the BER,NER ability to inhibit DNA repair,increased DNA damage,interfere DNA replication.