采用多药耐药基因mRNA转染人单个核血细胞提高对化疗药物耐药性的体外研究  

Transfection of MDR1-mRNA into human mononuclear cells to improve their resistance to anticancer agents, an in vitro study

作　　者：李雷[1] 向阳[1] 田芳[2] 杨秀玉[1] 任彤[1] 

机构地区：[1]中国医学科学院中国协和医科大学北京协和医院妇产科 [2]军事医学科学院肿瘤分子实验室

出　　处：《中华医学杂志》2002年第12期848-851,共4页National Medical Journal of China

摘　　要：目的　采用将人多药耐药基因 (MDR1)mRNA导入人单个核血细胞的方法 ,观察跨膜糖蛋白 (P gp)在转染后单个核血细胞中的表达和功能情况 ,并初步探讨此方法中影响P gp表达和功能的因素。方法　体外分别合成无 5′和 3′UTR(非翻译区 )的MDR1mRNA[pGEM5Zf(+) MDR1]和使用Xenopusβ globin的 5′和 3′UTR替代野生型 5′和 3′UTR的MDR1mRNA(pT7TS MDR1) ,从而构成两个试验组。然后利用脂质体为介质转染富含造血细胞的人单个核血细胞 ,使用流式细胞技术对MDR1基因的翻译产物P gp的表达和功能进行检测。结果　转染后 12h人单个核血细胞P gp的表达 :阴性对照组为 2 16 % ,pGEM5Zf(+) MDR1组为 8 94 % ,pT7TS MDR1组为 19 14 % ;转染后 72h人单个核血细胞P gp的表达 :阴性对照组为 2 12 % ,pGEM5Zf(+) MDR1组为 6 12 % ,pT7TS MDR1组为 10 71%。转染后 12h的Rhodamine 12 3排泌实验 :阴性对照组为 19 2 0 % ,pGEM5Zf(+) MDR1组为 2 5 2 9% ,pT7TS MDR1组为 35 0 2 %。结论　转染后 12h及 72h两试验组都表现出P gp的高表达 ,并且以pT7TS MDR1组为优 ;转染后细胞对P gp的表达及存在可以维持至少 72h ;表达的P gp具有外排泵功能。Objective To investigate the expression and function of P glycoprotein (P gp), an efflux pump encoded by multidrug resistance complementary DNA (MDR1), in human mononuclear cells (MNCs) transfected with MDR1 mRNA. Method Two kinds of human MDR1 mRNA, pT7TS MDR1 and pGEM5Zf (+) MDR1 with difference in the 5' and 3' untranslated regions only, were constructed and modified and then were transfected into human MNCs rich in hematopoietic progenitor cells from a patient with small cell lung cancer. The expression efficiency and pump function of P gp were measured with FACS. Rhodomine 123 efflux test was used to examine the function of P gp. Un transfected mononuclear cells from the same person were used as controls. Result The expression of P gp 12 hours after transfection was 2.16% in control group, 8.94% in pGEM5Zf (+) MDR1 group, and 19.14% in pT7TS MDR1 group; The expression of P gp 72 hours after transfection was 2.12% in control group, 6.12% in pGEM5Zf(+) MDR1 group, and 10.71% in pT7TS MDR1 group, significantly higher in the third group. Rhodamine 123 efflux test showed that the efflux pump function of P gp 12 hours after the trransfection was 19.20% in control group, 25.59% in GEM5Zf (+) MDR1 group, and 35.02% in pT7TS MDR1 group (all P <0.01). The expression and presence of P gp by transfected cells lasted at least 72 hours. Conclusion Transfection of human mononuclear cells with MDR1 mRNA significantly increases their resistance to anticancer agents.

关 键 词：多药耐药基因 MRNA 化疗药物 体外研究 单个核血细胞 妇科肿瘤 

分 类 号：R730.53[医药卫生—肿瘤]