MDR1特异性核酶逆转肝癌多药耐药的实验研究  

MDR1 specific ribozyme reverses multidrug resistance (MDR) of human hepatocellular carcinoma, an in vivo experimental study

作　　者：郜永顺[1] 陈孝平[2] 张水军[1] 丁磊[2] 李开艳[3] 吴在德[2] 

机构地区：[1]郑州大学第一附属医院普通外科,450052 [2]华中科技大学同济医学院附属同济医院肝脏外科中心 [3]华中科技大学同济医学院附属同济医院B超室

出　　处：《中华普通外科杂志》2006年第12期882-885,共4页Chinese Journal of General Surgery

基　　金：卫生部临床学科重点基金资助项目(2001-321)卫生部科技专项基金资助项目(WKZ-2000-1-15)

摘　　要：目的探讨MDR1核酶(N2A+tRNAimet-iMDR1-sRz,sRz)在裸鼠体内逆转人肝癌组织多药耐药(MDR)的可行性。方法将原发性MDR人肝癌组织裸鼠原位移植模型第2代随机分为A组(空白对照组:生理盐水40μl+Lipofect AMINETM 2000 10μl)、B组(阴性对照组:N2A+tRNAimet 10μg／40μl+Lipofect AMINETM 2000 10μl)和C组(核酶组:sRz 10μg／40μl+LipofectAMINETM 2000 10μl),均开腹瘤内注射。瘤内注药1周后用表阿霉素15 ms／kg腹腔注射,每周1次,连续4周。彩色B超测量肿瘤体积。化疗结束后1周处死裸鼠,RT-PCR、Western blot法检测肿瘤中MDR1 mRNA及其蛋白P-gp的表达。结果C组每次化疗后肿瘤体积均较前缩小(F=659．99,P<0．05)。除第1次化疗外,其余各次化疗后C组的抑制率均高于A、B组(F=35．36,12．77,97．60,P<0．05)。化疗结束后,C组与瘤源以及A、B组相比,肿瘤组织中MDR1 mRNA和P-gP的表达明显降低(F= 45．36,3590．40,P<0．05)。结论sRz可有效降低肝癌细胞表达MDR1 mRNA和P-gp,一定程度上逆转MDR,提高E-ADM的化疗效果。单纯E-ADM化疗可使肝癌组织MDR1 mRNA和P-gp表达升高,诱导MDR的产生。Objective To investigate the reversal of MDR by using anti-MDR1 ribozyme N2A ＋ tRNAi^met-iMDR1- sRz （sRz） in nude mice bearing human hepatocellular carcinoma. Methods The nude mice model with implanted human hepatocellular carcinoma was randomly devided into group A （ saline 40 μl ＋ Lipofect AMINE^TM2000 10 μ1）, B （N2A ＋ tRNAi^met 10 μg/40 μl ＋ Lipofect AMINE^TM2000 10 μl ） and C（ sRz 10 μg/40 μl ＋ Lipofect AMINE^TM2000 10 μl）. After one week, mice was peritoneally injected E-ADM 15 mg · kg^-1 once a week for 4 weeks. The size of tumors was measured with B-ultrasound and the tumor inhibition rate was calculated. One week after chemotherapy mice was sacrificed and MDR1 mRNA and P-gp were investigated with RT-PCR and Western blot. Results In group C tumor shrank upon each chemotherapy （ F = 659. 99, P 〈 0. 05 ）. The tumor inhibition rate in group C was higher than that in group A and B（F =35. 36, 12. 77, 97.60, P 〈0. 05）. MDR1 mRNA and P-gp expressions were down regulated in group C than group A, group B and that in original tumor （ F = 45.36, 3590. 40, P 〈 0.05 ）. Conclusions E-ADM up-regulates the expressions of MDR1 mRNA and P-gp. Anti-MDR1 ribozyme sRz efficiently down-regulates the expressions of MDR1 mRNA and P-gp, and reverses MDR.

关 键 词：癌 肝细胞 多药耐药相关蛋白类 核酶 裸鼠 

分 类 号：R735.7[医药卫生—肿瘤]