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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
【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.
- 【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