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目的:评估血红素加氧酶(HO)对肝癌耐药细胞化疗敏感性的影响,探讨HO对肝癌细胞多药耐药性可能的影响机制。方法:血红素加氧酶抑制剂锌卟啉(ZnPP)和诱导剂Hemin作用于Bel/Fu肝癌耐药细胞株,分别应用MTT、RT-PCR和流式细胞术检测细胞株对化疗药物敏感性、血红素加氧酶-1(HO-1)及多药耐药基因(MDR-1)核酸水平表达量与P-糖蛋白(P-GP)功能。结果:Hemin作用于Bel/Fu细胞诱导24 h后,化疗药物半数抑制浓度明显增加(P<0.01),HO-1 mRNA表达明显增加(F=71.513,P<0.05),MDR-1 mRNA的表达量显著增加(F=2 340,P<0.01),且呈剂量依赖趋势,ZnPP干预组表现则相反;MDR-1cDNA/β-actin的表达量呈现与HO-1cDNA/β-actin平行变化的趋势,两者呈直线正相关(r=0.992,a=-0.044,b=1.223);Hemin组罗丹明荧光曲线明显左移,且浓度越高曲线左移越明显,而ZnPP组则表现为曲线右移,出现相反的结果。结论:影响HO-1的表达可改变肝癌耐药细胞株的化疗敏感性,这种变化是通过影响MDR-1表达,改变P-GP药物外排功能实现的;HO-1可作为逆转肿瘤多药耐药的潜在作用靶点。
Objective: To evaluate the effect of heme oxygenase (HO) on the chemosensitivity of drug-resistant hepatocarcinoma cells and to explore the possible mechanism of HO on multidrug resistance of hepatoma cells. Methods: Heme oxygenase (ZnPP) and Hemin were used to induce drug resistance in Bel / Fu hepatocarcinoma cell lines. MTT, RT-PCR and flow cytometry were used to detect the chemosensitivity , Heme oxygenase-1 (HO-1) and multidrug resistance gene (MDR-1) and P-glycoprotein (P-GP) Results: After Hemin treated with Bel / Fu cells for 24 h, the half inhibitory concentration of chemotherapeutics significantly increased (P <0.01), the expression of HO-1 mRNA increased significantly (F = 71.513, P <0.05) The expression of MDR-1 cDNA / β-actin showed a parallel trend with that of HO-1 cDNA / β-actin in a dose-dependent manner (F = 2 340, P <0.01) (R = 0.992, a = -0.044, b = 1.223). The rhodamine fluorescence curve of Hemin group shifted significantly to the left, and the higher the concentration, the more obvious the curve shifted to the left, while the ZnPP group showed a curve right Move, the opposite result. CONCLUSION: The effect of HO-1 expression on chemosensitivity of drug-resistant hepatocarcinoma cell lines can be achieved by affecting the expression of MDR-1 and changing the drug efflux function of P-GP. HO-1 can be used as a reverse multi-tumor Drug resistance potential target.