凋亡抑制基因c-FLIPs对人肺腺癌细胞增殖和凋亡的作用

摘要

Objective:To evaluate the effects of recombinant adenovirus apoptosis inhibiting gene c - FLIPs on proliferation and apoptosis of human lung adenocarcinoma cells. Methods:Recombinant adenovirus c - FLIPs was constructed and transfected to human lung adenocarcinoma cells Calu - 3. Real - time PCR detected c - FLIPs, Caspase - 8,Caspase - 10 and Bcl - 2,mononuclear cell direct cytotoxicity assay for cell proliferation,and flow cytom-etry for apoptotic after transfected. Results:Successfully constructed the over expression of recombinant adenoviral Ad5 c - FLIPs. Real - time PCR detected c - FLIPs expression in Calu - 3 cell with high expression,Caspase - 8, Caspase - 10 and Bcl - 2 expression with low expression. MTT assay found that over expression of c - FLIPs gene can induce cell proliferation. Apoptosis rate in control group(55. 17 ± 9. 68)% was higher than Calu - 3 cells(10. 97 ± 1. 66)% ,P < 0. 05. Conclusion:c - FLIPs can significantly induce the proliferation of human lung adenocarcinoma cells,and inhibit the apoptosis.%目的：探讨过表达凋亡抑制基因 c - FLIPs 重组腺病毒对人肺腺癌细胞增殖和凋亡的作用。方法：构建过表达凋亡抑制 c - FLIPs 的重组腺病毒 Ad5 c - FLIPs，感染人肺腺癌细胞 Calu -3。Real - time PCR 检测感染前后 c - FLIPs、Caspase -8，Caspase -10和 Bcl -2的表达。MTT 法检测细胞增殖。流式细胞仪检测感染Ad5 c - FLIPs 后人肺腺癌细胞的凋亡情况。结果：成功构建过表达 c - FLIPs 重组腺病毒 Ad5 c - FLIPs，real- time PCR 检测凋亡抑制基因 c - FLIPs 在 Calu -3细胞株高表达，过表达 c - FLIPs 基因，凋亡相关基因Caspase -8、Caspase -10和 Bcl -2的表达明显降低。MTT 法检测感染前后细胞增殖情况发现，过表达 c -FLIPs 基因可诱导细胞增殖，流式细胞仪分析经 PI 染色后的 Calu -3细胞株，对照组和腺病毒感染组细胞的凋亡率分别为（55.17±9.68）%和（10.97±1.66）%，差异具有统计学意义（P <0.05）。结论：凋亡抑制基因c - FLIPs 可明显诱导人肺腺癌细胞增殖并抑制凋亡。