p27kip1基因在人胆管癌细胞化疗增敏效应中的作用

目的 探讨外源性p27^kip1在人胆管癌细胞系QBC939中高表达对胆管癌细胞化疗敏感性的影响。方法 通过携带人p27^kip1基因的重组腺病毒载体Ad-p27mt转染人胆管癌细胞系QBC939。经逆转录-聚合酶链反应（RT-PCR）、Western blot检测p27^kip1在胆管癌细胞中的表达；经MTT比色法及流式细胞术检测5-氟尿嘧啶（5-FU）、丝裂霉素（MMC）、环磷酰胺（CTX）对转染p27^kip1前后的QBC939细胞生长抑制及凋亡的影响。结果 Ad-p27mt转染后5-FU、MMC和CTX对QBC939细胞生长抑制率，分别由转染前的41．89％、45．59％和38．91％显著增加为56．15％、55．65％和51．69％；凋亡率也由13．76％、11．76％和10．46％，明显升高为41．39％、35．94％、34．46％，差异均有显著性意义（均P〈0．05）。结论 p27^kip1在QBC939细胞中高表达，能显著增强胆管癌细胞对化疗药物的敏感性，为基因联合化疗药物治疗胆管癌提供了参考依据。

Sensitivities of Cholangiocarcinoma Cells Transfected by Exogenous p27kip1 Gene to Chemotherapy

Objective To investigate the effects of exogenous p27~(kip1) gene on chemosensitivity of human cholangiocarcinoma cell line.Methods The recombinant vector was constructed and the mutated p27~(kip1) gene was transfected into human cholangiocarcinoma cell line(QBC_(939)).RT-PCR and Western blot were used to determine the expression of target genes.The effects of 5-fluorouracil(5-FU),mitomycin(MMC) and cyclophosphamide (CTX) on the transfected cells were detected by assaying the rate of apoptosis and growth inhibition by methabenzthiazuron(MTT) assay and flow cytometry(FCM). Results The exogenous p27~(kip1)gene was expressed effectively in the cells,and the expression enhanced the apoptosis and growth inhibition of QBC_(939) induced by 5-FU,MMC and CTX.The ratio of growth inhibition was increased significantly from 41.89 %(5-FU),45.59 %(MMC) and 38.91 %(CTX) to 56.15 %(5-FU),55.65 %(MMC) and 51.69 %(CTX) and apoptosis index from 13.76 %(5-FU),11.76 %(MMC),10.46 %(CTX) to 41.39 %(5-FU),35.94 %(MMC),34.46 %(CTX) respectively.Conclusion The exogenous p27~(kip1) gene transfer can remarkably increase the drug sensibility of the cholangiocarcinoma cells.The combination of p27~(kip1) gene therapy with chemotherapy may be more effective in cancer treatment.