摘要
背景与目的在中国肺癌居各种恶性肿瘤之首位,寻求肺癌的基因疗法已成为人们努力的方向。本研究拟探讨生长终止特异性同源盒基因(growtharrest-specifichomeobox,Gax)对A549细胞系增殖及其原癌基因c-fos、c-junmRNA表达的影响。方法以腺病毒介导Gax基因(adenoviraltransductionoftheGaxgene,Ad-Gax)转染A549细胞,RT-PCR和免疫细胞化学法测定转染前后细胞中GaxmRNA和蛋白表达;RT-PCR测定转染前后细胞中c-fos、c-junmRNA表达变化;MTT法检测Gax基因转染对A549细胞生长的抑制作用,计算24、48、72h细胞抑制率。结果RT-PCR和免疫细胞化学证实Gax基因转染A549细胞后Gax能稳定表达,未转染组未发现Gax表达;RT-PCR证实转染组A549细胞中c-fos、c-junmRNA比未转染组显著降低(t=7.755,P<0.01;t=5.938,P<0.01);MTT法检测显示转染组A549细胞在24、48、72h的抑制率分别是(47.35±5.36)%、(54.96±1.78)%、(65.39±5.11)%,随时间逐渐升高与未转染组同时间比较有显著性差异(χ2=7.152、9.431、12.847,P<0.01)。结论Gax基因转染能抑制A549细胞的增殖,其分子机制可能与Gax下调c-fos、c-jun的表达有关。
Background and objective Lung cancer is the leading cancer of malignant tumor in China. It is the direction that poeple make efforts to seek gene therapy of lung cancer. The aim of this study is to explore the effects of transfected growth arrest-specific homeobox gene （Gax gene） on the proliferation and expressions of c-fos and c-jun mRNA in A549 cells. Methods A549 cells were transfected with Gax gene by adenovirus. Expressions of Gax mRNA and protein were detected by RT-PCR and immunocytochemistry. The expressions of c-fos and c-jun mRNA were evaluated by RT-PCR. The proliferation inhibition effect of Gax transfection on A549 cells was evaluated by MTT assay. Results Only in the A549 cells transfected with Gax gene the Gax expression was confirmed by RT-PCR and immunocytochemistry. Compared with that in the control group, c-fos and c-jun mRNA level decreased significantly in Gax-transfected A549 cells （t= 7. 755, P〈0. 01; t = 5. 938, P〈0.01）. MTT assay showed that the proliferation inhibition rates of A549 cells transfected by Ad-Gax for 24 h, 48h and 72h were （47.35±5.36）%, （54. 96±1. 78）%, and （65.39±5.1 1）% respectively. And these proliferation inhibition rates were significantly higher than those in the control group （X^2= 7. 152, 9. 431 and 12. 847, P〈0.01）. Conclusion Gax gene can inhibit the proliferation of A549 cells. Its molecular mechanism may be through down regulating the expressions of c-fos and c-jun.
出处
《中国肺癌杂志》
CAS
2006年第3期221-225,共5页
Chinese Journal of Lung Cancer
