醛糖还原酶相似基因转染细胞对含碳酰基抗癌药物耐药的研究  

Establishment and identification of the gene cell line transfected aldose reductase like gene and its drug resistance to drugs containing carbonyl group

作　　者：王建[1] 万腊香[1] 吴孟津[1] 万载阳[1] 危当恒[1] 杨永宗[1] 

机构地区：[1]湖南南华大学分子生物中心,衡阳421001

出　　处：《中华肝脏病杂志》2002年第6期445-448,共4页Chinese Journal of Hepatology

基　　金：国家自然科学基金(C03020705)

摘　　要：目的 研究醛糖还原酶相似蛋白(aldose reductase like prptein,ARL-1)在肝癌细胞中的功能及ARL-1基因高表达与肝癌耐药之间的关系。方法 通过阳离子脂质体转染法将ARL-1基因导入HepG2细胞,建立稳定表达该基因的细胞株,采用MTT法检测细胞对含醛基药物的耐药变化情况。结果 转染细胞与未转染细胞相比,对含醛基的抗肿瘤药物如阿霉素、丝裂霉素的耐药性明显增强,其半数抑制浓度(IC50)分别提高到2.6倍和3.1倍,ADM组t=6.39,P<0.05,MMC组t=30.06,P<0.01。用不含醛基的5-氟尿嘧啶处理两组细胞,则未发现有明显的耐药差异(t=0.684,P>0.05)。结论 ARL-1的表达上调可能是肿瘤细胞对含醛基抗肿瘤药物耐药的主要原因之一。Objective To investigate the relationship between aldose reductase like protein (ARL-1) gene overexpressed in HCC cells and drug-resistance of the cell to drugs containing carbonyl group. Methods To establish ARL-1 stable expression positive cell line, eukaryotic expression vectors containing ARL-1 gene cDNA were transfected into Hep cell mediated by lipofect AMINE. The positive monoclones were determined by PCR and RT-PCR, respectively. Then MTT assay was used to study the drug resistance ability of the cells to drugs containing carbonyl after incubating three days with those drugs. Results After ARL-1 gene transfection mediated by lipofect AMINE, one positive monoclonal cell overexpressing ARL-1 gene was selected. Compared with the control cell group, drug resistance ability of the positive cells to ADM and MMC which contain carbonyl group increased 2.3 and 3.17 fold, respectively (t=6.39, P=0.016 in ADM group and r=30.06, P=0.001 in MMC group). In the same time, drug resistance ability to 5-FU which has no carbonyl group had no statistical difference between positive monoclonal cell group and control cell group (t=0.684, P=0.531). Conclusions The Hep ARL-1 positive cell line with stable expression of ARL-1 gene has been established successfully and the up-regulation of ARL-1 gene may plays an important role in drug resistance of the cells to anticancer drugs containing carbonyl group.

关 键 词：碳酰基 醛糖还原酶相似蛋白 耐药 肝癌 细胞转染 

分 类 号：R73-36[医药卫生—肿瘤]