多药耐药基因转染的CIK细胞耐药性研究  

Study on multidrug resistance to tumor of cytokine-induced killer(CIK) cells after transfected with the multidrug resistance(mdr1) cDNA

作　　者：李淑艳[1] 王淑英[1] 高涵[1] 冯丽[1] 富东娜[1] 师岩[1] 吴琦[1] 郭红艳[1] 齐晓丹[1] 

机构地区：[1]齐齐哈尔医学院生物化学教研室,黑龙江齐齐哈尔161006

出　　处：《中国医药导报》2009年第35期27-29,共3页China Medical Herald

基　　金：黑龙江省教育厅基金资助项目(11541406)

摘　　要：目的:将多药耐药基因(mdr1)转入CIK细胞,观察转染前后其对顺铂的耐药性及对Lewis肺癌细胞杀伤活性的影响。方法:常规方法培养CIK细胞,在细胞对数生长期,将mdr1的重组质粒转染CIK,通过RT-PCR鉴定耐药基因表达;MTT法检测CIK细胞对顺铂敏感性的变化,同时检测转染前后CIK细胞对Lewis肺癌细胞的杀伤活性变化;Western blot检测细胞中mdr1编码的P-gp蛋白表达的变化;通过计算瘤重抑制率(TWI)检测转染前后CIK细胞对Lewis肺癌移植瘤的抑制作用。结果:转染mdr1后的CIK细胞mdr1 mRNA阳性,在转染后的CIK细胞中,P-gp的表达较转染前CIK细胞及转染空质粒的CIK细胞明显增高,差异有统计学意义(P<0.05),转染mdr1基因后的CIK细胞对顺铂的耐药性明显提高,转染前后CIK细胞对Lewis肺癌细胞的杀伤活性无明显变化(P>0.05)。结论:将mdr1基因转入CIK细胞后,细胞获得了多药耐药性,同时保持了原有的对肿瘤细胞的杀伤活性。Objective： To investigate the muhidrug resistance to Cisplatin and remaining cytotoxic activity to Lewis lung carcinoma cell line of cytokine-induced killer （CIK） cells after transfected with the muhidrug resistance （mdr1） cDNA. Methods： CIK cells were induced by culturing PBMC with regular method. CIK cells were transfected with human mdr1 cDNA recombinant plasmid, RT-PCR was used to detect mdr1 mRNA in transfected CIK cells. Muhidrug resistance to Cisplatin and cytotoxic activity to Lewis lung carcinoma cell line were performed using MTT method. P-glycoprotein （P-gp） expressed of CIK cells was assayed by Western blot. The inhibitory effect of transfected CIK cells on Lewis lung carcinoma cell line was measured by the inhibition rate of solid tumor weight. Results： Mdrl mRNA was positive in transfected CIK cells, Pgp was highly expressed in the transfected CIK cells, as compared with those of untransfected CIK cells （P〈0.05）; the multidrug resistance to Cisplatin increased, however, the cytotoxic activity to Lewis lung carcinoma cell line remained unaltered （P〉0.05） in the transfected CIK cells. Conclusion： The success of transfected with mdr1 cDNA CIK cells has the characteristics of muhidrug resistance without change in their cytotoxic activity to tumor cells.

关 键 词：细胞因子诱导的杀伤 mdr1 多药耐药 基因转染 

分 类 号：R392.1[医药卫生—免疫学]