短发夹RNA/MDR1逆转肝细胞癌多药耐药  

Reversal of multidrug resistance of HepG2/MDR1 by shRNA/MDR1 in vivo

作　　者：潘光栋[1] 严律南[2] 王新平[2] 夏冬[2] 夏庆杰[2] 闫乃红[2] 陈清英[2] 

机构地区：[1]广西医科大学第五附属医院肝胆外科,柳州545001 [2]四川大学华西医院

出　　处：《中华实验外科杂志》2007年第12期1511-1513,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金(30170925)

摘　　要：目的探讨短发夹 RNA/MDR1(shRNA/MDR1)干扰技术能否体内逆转肝细胞癌多药耐药。方法建立肝细胞癌耐药细胞株 HepG2/MDR1和敏感细胞株 HepG2裸鼠移植瘤模型,分腹腔注射组和瘤体内注射组,以表达载体 pSUPER-shRNA/MDR1转染,阴性对照组以阴性载体pSUPER 转染,72 h 后行腹腔内阿霉素化疗试验,每周2次,共2周,然后处死动物,测量化疗前后肿瘤体积差,瘤体制成细胞悬液和组织切片,分别以流式细胞术和免疫组织化学检测细胞膜 P-gp 表达。结果成瘤率100%,HepG2/MDR1组和 HepG2组成瘤时间和化疗前肿瘤体积差异无统计学意义,HepG2/MDR1组化疗前后肿瘤体积差(mm^3)与阴性对照组比较差异有统计学意义(700.14±25.61比1659.70±152.54,P<0.01);腹腔注射组和瘤内注射组差异无统计学意义。流式细胞术检测发现治疗组细胞膜蛋白 P-gp 表达率(%)较阴性对照组明显下调(65.1%比94.1%,P<0.05),腹腔注射组和瘤内注射组差异无统计学意义(94.1%比92.8%,P>0.05)。瘤体组织病理切片和免疫组织化学分析也有同样的结果。结论表达载体 pSUPER-shRNA/MDR1体内转染可以逆转肝细胞癌多药耐药。Objective To explore the possibility of reversal of multidrug resistance of HepG2/ MDR1 by shRNA/MDR1 in vivo. Methods The implant tumors of nude mice were established by injection of cells HepG2/MDR1 and HepG2. Tumors in HepG2/MDR1 were divided into in situ injection groups and intraperitoneal injection groups and were transfeeted by pSUPER-shRNA/MDR1. Negative groups were transfeeted by pSUPER-shRNA/MDR1 negative vectors. Seventy-two h after transfeetion, adriamyein was injected into peritoneal cavity twice a week for two weeks. Tumors were collected by executing the mice to make cell suspension and histological sections for detecting the expression of P-glyeoprotein （P-gp） by flow cytometry and immunohistochemistry. Results The tmnorigenic rate was 100%. There was no difference in tumorigenic time and tumor volume before-chemotherapy between HepG2/MDR1 and HepG2. Compared to negative controls,the increase of tumor volume after chemotherapy in HepG2/MDR1 was slower （ 1659.70 ± 152.54 vs 700.14 ±25.61 ,P 〈 0.01 ）. The expression of P-gp in HepG2/MDR1 was lower than that of negative controls （ 65.1% vs 94.1%, P 〈 0.05 ）. There was no difference between in situ injection groups and intraperitoneal injection groups. Similar results were found by immunohistochemistry. Conclusion The multidrug resistance of HepG2/MDR1 could be reversed by shRNA/MDR1 in v/vo.

关 键 词：癌 肝细胞 多药耐药 RNA干扰 

分 类 号：R735.7[医药卫生—肿瘤]