瘤体内注射逆转录病毒上清液介导体内基因导入

Gene in retrovirus supernatant was transfered by injecting intratumorly in nude mice

目的 探讨逆转录病毒上清液瘤内及瘤周注射介导体内基因导入。方法 应用逆转录病毒上清液裸鼠皮下移植瘤内及瘤周注射法导入多药耐药基因 (mdr1 ) ,腹腔注射化疗药物观察肿瘤模型的耐药性 ,免疫组化检测肿瘤细胞P 糖蛋白的表达 ,RT PCR检测肿瘤内mdr1mRNA的表达量。结果 携带mdr1基因的逆转录病毒上清液注射后肿瘤表现出多药耐药性 ,高滴度病毒上清液可达到 60 %细胞表达P GP ;RT PCR表明 ,基因转移后肿瘤组织内mdr1mRNA的表达量明显增加。结论 逆转录病毒上清液瘤内及瘤周注射 ,可发生部分肿瘤细胞的基因转移 。

Objective In order to study gene transfer in vivo,supernatant of retrovirus carrying an mdr1 cDNA was injected intratumorly and subcutaneously around the tumor in nude mice.Methods Retrovirus supernatant was injected intratumorly and subcutaneously around the tumor.Multidrug resistance of tumor model was measured by intraperitoneal injecting adriamycin 5mg/kg or colchicine 500ng/kg;expression of P glycoprotein (P GP) on membrane and mdr1 mRNA in cells was detected with immunocytochemistry and reverse transcription polymerase chain reaction (RT PCR).Results Multidrug resistance can be showed in tumor model after injecting retrovirus supernatant with mdr1 gene.Immunocytochemistry assay with MCAb against P GP of tumor tissue was examined,comparable high levels of P GP expression was observed,and efficiency of strong positive cells of high titer virus was 60%;Mdr1 mRNA detected by RT PCR was significantly increased in injection group as compared with control one.Conclusion Gene transfer can be efficiently developed by injecting of retrovirus supernatant intratumorly and subcutaneously around the tumor,high titer retrovirus transfering more efficiently than low one.