甲基莲心碱逆转人结肠癌细胞奥沙利铂耐药的体外研究

发布时间：2018-11-22 12:52

【摘要】：目的:探讨甲基莲心碱(Nef)对人结肠癌细胞奥沙利铂(OXA)耐药的逆转作用及机制。方法:采用OXA浓度逐步递增法(2、4、8、12、24、48μmol/L)孵育人结肠癌HCT116细胞诱导构建OXA耐药株HCT116/OXA;检测Nef对HCT116/OXA细胞的细胞毒性,确定Nef的最适作用浓度和时间;分析并比较OXA(IC_(50)浓度)单独处理、Nef(最适作用浓度)单独处理、OXA(IC_(50)浓度)联合Nef(最适作用浓度)处理后,HCT116/OXA细胞的增殖,凋亡情况及凋亡相关蛋白(Bcl-2,Bax,PARP,p-PARP)的表达情况。结果:与亲本HCT116细胞比较,HCT116/OXA细胞较对OXA的耐药性明显增高(IC_(50):21.00μmol/Lvs.112.00μmol/L,P0.05),耐药指数为5.33。Nef能明显抑制HCT116/OXA的增殖有作用(P0.05),并呈浓度依赖性,其最适作用浓度、时间分别为5μmol/L、24h(细胞存活率为90%)。与OXA单独处理比较,HCT116/OXA细胞对OXA联合Nef处理的耐受性明显降低(IC_(50):112.00μmol/Lvs.45.47μmol/L,P0.05),逆转倍数为2.46;Nef单独作用对HCT116/OXA细胞的凋亡影响不明显(P0.05),但其与OXA联合作用对HCT116/OXA细胞凋亡诱导作用明显强于OXA单独作用(P0.05);与OXA或Nef单独作用比较,OXA联合Nef作用后,HCT116/OXA细胞抗凋亡蛋白Bcl-2表达明显下降,Bax、p-PARP等凋亡蛋白表达明显上升(均P0.05)。结论:Nef可逆转HCT116/OXA对OXA的耐药,机制可能与其调节Bcl-2/Bax表达水平,从而与OXA产生协同作用有关。
[Abstract]:Aim: to investigate the reversal effect of (Nef) on oxaliplatin (OXA) resistance in human colon cancer cells and its mechanism. Methods: human colon cancer HCT116 cells were incubated with the method of increasing the concentration of OXA (2? 4? 48 渭 mol/L). The OXA resistant cell line HCT116/OXA; was used to detect The proliferation of HCT116/OXA cells treated with OXA (IC_ (50) concentration alone), OXA (IC_ (50) concentration alone and Nef (optimal concentration) was analyzed and compared. Apoptosis and expression of apoptosis-related protein (Bcl-2,Bax,PARP,p-PARP). Results: compared with parental HCT116 cells, the resistance of HCT116/OXA cells to OXA was significantly higher than that of HCT116/OXA cells (IC_ (50): 21.00 渭 mol/Lvs.112.00 渭 mol/L,P0.05). The drug resistance index (5.33.Nef) could significantly inhibit the proliferation of HCT116/OXA (P0.05) in a concentration-dependent manner, and the optimal concentration was 5 渭 mol/L,24h (cell survival rate was 90%). Compared with OXA alone, the tolerance of HCT116/OXA cells to OXA combined with Nef was significantly decreased (IC_ (50): 112.00 渭 mol/Lvs.45.47 渭 mol/L,P0.05), and the reversal multiple was 2.46; The effect of Nef alone on the apoptosis of HCT116/OXA cells was not significant (P0.05), but the combined effect of Nef and OXA on the apoptosis of HCT116/OXA cells was significantly stronger than that of OXA alone (P0.05). Compared with OXA or Nef alone, OXA combined with Nef significantly decreased the expression of anti-apoptotic protein Bcl-2 and increased the expression of Bax,p-PARP and other apoptotic proteins in HCT116/OXA cells (P0.05). Conclusion: Nef can reverse the drug resistance of HCT116/OXA to OXA, and the mechanism may be related to the regulation of Bcl-2/Bax expression and the synergistic effect of OXA.
【作者单位】：中南大学湘雅医院医学科学研究中心;中南大学湘雅医院肿瘤科;湖南省分子放射肿瘤学重点实验室;
【基金】：国家自然科学基金资助项目(30770970;81172471;81070362;81372629) 湖南省自然科学基金重点资助项目(11JJ2049)
【分类号】：R735.35

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