基因修饰对消化道肿瘤细胞化疗药敏感性的影响

Retrovirus-mediated Transduction of Uracil Phosphoribosyltransferase Gene Render Alimentary Tumor Cells More Sensitive to 5-Fluorouracil in Vitro

目的:探讨UPRT基因修饰消化道肿瘤细胞后对5-氟脲嘧啶(5-FU)杀伤效应的增强作用。方法:采用PCR技术从大肠杆菌K12菌株基因组中扩增UPRT基因,并构建pLXSN逆转录病毒表达载体,进一步转染人胃癌细胞株SCG7901、大肠癌细胞株Lovo以及肝癌细胞株7721。采用MTT法检测转导UPRT基因前后,肿瘤细胞对5-FU敏感性的变化。结果:通过逆转录病毒载体介导UPRT基因修饰肿瘤细胞,体外MTT法检测结果显示,转导UPRT基因后可使SCG7901、Lovo及7721对5-FU的敏感性分别提高约21.07、30.25及35.19倍。结论:UPRT基因修饰肿瘤细胞,能明显提高其对5-FU杀伤的敏感性,增强5-FU的抗瘤效应。

Objective:To investigate the killing effects of UPRT/5-FU Enzyme /Prodrug System to alimentary tumor cells.Methods:The UPRT gene encoding uracil phosphoribosyltrans-ferase was amplified from E.Coli K12genome and subcloned into retrovirus expression vec tor pLXSN,Recombinant retrovirus was used further to infect human gastric cancer cell line SCG7901、colon cancer cell line Lovo and hepatic cancer cell line7721.The sensitivity of tumor cells transduced with UPRT gene to5-FU in vitro was detected by MTT method.Results:Trans-duction of UPRT gene into SCG7901、Lovo and7721cell lines can render them more sensitive to5-FU,which increased with21.07-fold、30.25-fold and35.19-fold independently.Conclusion:Transduction of UPRT gene can render the tumor cell lines be more sensitive to low concentra-tion of5-FU and improve significantly the antitumor effect of5-FU in vitro.