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逆转录病毒转染法建立兔VX-2多药耐药株

Construction of MDR rabbit VX-2 cell line by retrovirus infection

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【作者】 樊爱琳刘国鹏郑善銮马静王执民丁振若杨麦贵郝晓柯

【Author】 FAN Ai Lin 1, LIU Guo Peng 2, ZHENG Shan Luan 1, MA Jing 3, WANG Zhi Min 4, DING Zhen Ruo 1, YANG Mai Gui 1, HAO Xiao Ke 1 1Department of Clinical Laboratory, Xijing Hospital, 2Center of Basic Medical Teaching Experiment, School of Basic Medicine, 3Department of Chinese Traditional Medicine, Xijing Hospital, Fourth Military Medical University, Xi’an 710033, China, 4Department of Interventional Radiology, Tangdu Hospital, Fourth Military Medical University, Xi’an 710038, China

【机构】 第四军医大学西京医院检验科第四军医大学基础部教学实验中心第四军医大学西京医院中医科第四军医大学唐都医院介入放射科第四军医大学西京医院检验科 陕西西安710033陕西西安710033陕西西安710038陕西西安710033

【摘要】 目的 :建立VX 2多药耐药 (MDR )细胞 (VX 2mdr1 ) .方法 :利用逆转录病毒转染法将带有mdr1cDNA全序列的逆转录病毒载体 pHaMDR1转入到兔VX 2细胞中 ,MTT法检测细胞在不同化疗药物作用下的存活率 ;免疫组化检测细胞的P 糖蛋白 (P gp)表达 ,RT PCR检测细胞内mdr1mRNA的表达量 .结果 :转基因的细胞对吡柔比星、秋水仙素的耐药性分别提高 1 0和 2 5倍 ,免疫组化见转基因细胞中P gp表达增加 ,RT PCR示细胞内mdr1mRNA的表达量增加 .结论 :利用逆转录病毒转染法构建了兔VX 2MDR细胞

【Abstract】 AIM: To establish a multidrug resistance (MDR) rabbit VX 2 (VX 2 mdr 1) cell line. METHODS: Rabbit VX 2 cells were transfected by mdr 1 retrovirus supernatant. MDR1 of rabbit cells were tested by MTT assay and the expression of P glycoprotein (P gp) on membrane and mdr 1 mRNA in cells was detected by immunocytochemistry and RT PCR. RESULTS: The rabbit VX 2 mdr 1 cells were found to be 9.5 times more resistant to pirarubicin and 25 times to colchicine than VX 2 cells. VX 2 mdr 1 cells expressed higher P gp than VX 2 cells on cell membrane and VX 2 mdr 1 cells showed much higher mdr 1 mRNA level than VX 2 cells. CONCLUSION: A MDR rabbit VX 2 cell line is successfully established by gene transfer.

【关键词】 VX-2细胞多药耐药逆转录病毒基因转移
【Key words】 VX 2 cellMDRretrovirusgene transfer
【基金】 陕西省科技攻关项目 (CX99A0 1 6)
  • 【文献出处】 第四军医大学学报 ,Journal of The Fourth Military Medical University , 编辑部邮箱 ,2003年23期
  • 【分类号】R73-36
  • 【被引频次】8
  • 【下载频次】55
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