摘要
目的为重组人p53腺病毒应用于临床非小细胞肺癌顺铂耐药患者提供实验依据。方法采用CCK-8试剂检测半数抑制浓度(IC50);应用流式细胞仪检测细胞周期分布及凋亡发生率;采用功能分类基因芯片检测药物作用前后的基因变化。结果顺铂联合重组人p53腺病毒后,H1299细胞顺铂的半数抑制浓度下降至5μg/mL(P<0.05),细胞的凋亡率上升为10.73%(P<0.05)。重组人p53腺病毒单药作用后,H1299细胞内p53基因大量表达。在两药联合作用下,导入的p53基因激活了由Fas/Apo-1/CD95系统、TNFR超家族、Bcl-2/Bax等所介导的凋亡通路,同时也上调了部分细胞周期调控基因。结论重组人p53腺病毒介导的基因治疗有希望成为治疗临床非小细胞肺癌顺铂耐药患者的一种有效的辅助手段。
Objective To provide experimental evidence of recombinant adenovirus-p53 （tAd-p53） as a gene therapy applied to non-small cell lung cancer （NSCLC） patients who were resistant to cisplatin. Methods Inhibitory concentration 50% （ IC50 ） was assayed by cell counting kit-8 （CCK-8） reagent. The changes of cell cycle and apoptosis rate were analyzed by flow cytometer. The Gene Array was used to elucidate the change of the genes. Results Combining with the tAd-p53, the IC50 of H1299 cells to cisplatin was 5 ug/mL（P 〈 0.05） , and the apoptosis rate rose to 10.73% （ P 〈 0.05 ）. Using the tAd-p53 only, the wild-type p53 gene was transferred into the H1299 cell line successfully. While using the eisplatin and tAd-p53 both, the p53 gene which had been transferred in the cell line activated the apoptosis mainly depending on Fas/Apo-1/CD95 system, TNFR supper family and Bcl-2/Bax, and at the same time, it also increased some cell-eycle regulating genes. Conclusion The rAd-p53 can become a promising adjuvant gene therapy of NSCLC patients who are resistant to cisplatin.
出处
《肿瘤基础与临床》
2008年第5期373-376,共4页
journal of basic and clinical oncology
关键词
重组人P53腺病毒
顺铂
耐药
肺肿瘤
recombinant adenovirus-p53
cisplatin
resistance
lung neoplasms
