腺病毒介导的鸟氨酸脱羧酶和S-腺苷甲硫氨酸脱羧酶双反义RNA抑制肺癌A-549细胞的实验研究

目的：探讨鸟氨酸脱羧酶（ODC）和S－腺苷甲硫氨酸脱羧酶（AdoMetDC）双反义重组腺病毒（Ad-ODC-AdoMetDCas）介导的ODC和AdoMetDC反义RNA对肺癌A 549细胞增殖和侵袭的抑制作用。方法：采用FACS检测Ad-GFP感染细胞中GFP表达，以测定不同MOI的腺病毒对肺癌A-549细胞的基因转染效率；应用MTT法观察Ad-ODC-AdoMetDCas对肺癌A-549细胞生长增殖的影响；采用Western印迹和HPLC的方法分别检测腺病毒对肺癌A-549细胞中ODC和AdoMetDC蛋白表达以及胞内多胺含量的抑制作用,并应用TUNEL标记检测法观察Ad-ODC-AdoMetDCas对肺癌细胞凋亡的影响。结果：基因转染效率结果显示，以50 MOI的Ad-GFP感染肺癌细胞48h后，大约有75% 肺癌A-549细胞GFP表达阳性；Ad-ODC-AdoMetDCas对肺癌A-549细胞生长增殖有明显抑制作用。Ad-ODC-AdoMetDCas可明显抑制肺癌细胞中ODC和AdoMetDC基因表达 ，基因抑制率大于60％（P＜0.05）。HPLC结果显示，肺癌A-549细胞感染Ad-ODC-AdoMetDCas后，细胞内三种多胺含量都明显降低（P＜0.05）。TUNEL标记检测结果证实，Ad-ODC-AdoMetDCas可引起肺癌A-549细胞明显凋亡。结论：ODC和AdoMetDC双反义腺病毒具有显著抑制肺癌增殖和侵袭的作用，对于肺癌的防治研究具有一定的意义，有望成为治疗肺癌的基因药物。

Inhibitory effect of ornithine decarboxylase and S-adenosylmethionine decarboxylase bi-antisense virus on lung cancer cell A-549

Objective: To study the inhibitory effect of ornithine decarboxylase(ODC) and S-adenosylmethionine decarboxylase bi-antisense AdoMetDCas(Ad-ODCAdoMetDCas) virus on polyamine biosynthesis,proliferation and invasion of lung cancer cells.Methods: Adenovirus-mediated gene transduction efficiency was assessed by counting positive GFP cells using FACS.The malignant phenotype of A-549 cells was assessed by growth curves.Western Blot and HPLC were used to determine the expression of ODC and AdoMetDC and the polyamine content in A-549 cells,respectively.TUNEL was used to analyze cell apoptosis.Results: Approximate 75% of A-549 cells were infected with Ad-ODC-AdoMetDCas when MOI reached 50.Expression of ODC was inhibited in the infected tumor cells.Ad-ODC-AdoMetDCas inhibited growth and invasive ability of A-549 cells.TUNEL proved that Ad-ODC-AdoMetDCas could lead to cell apoptosis.Ad-ODC-AdoMetDCas vector-mediated gene transfer inhibited tumor cell growth through the blockade of the polyamine synthesis pathway. Conclusion: Ad-ODC-AdoMetDCas has a significant inhibitory effect on proliferation and invasion of lung cancer cells and has therapeutic potential for the treatment of lung cancer.