鸟氨酸脱羧酶和S-腺苷甲硫氨酸脱羧酶双反义腺病毒对肺癌增殖和侵袭抑制作用的研究

Inhibitory effects of ODC and AdoMetDC biantisense virus on the growth and invasion of lung cancer cell A-549

目的探讨重组鸟氨酸脱羧酶和S-腺苷甲硫氨酸脱羧酶双反义腺病毒载体（Ad-ODC-AdoMetDCas）介导的鸟氨酸脱羧酶（ODC）和S-腺苷甲硫氨酸脱羧酶（AdoMetDC）反义RNA对肺癌细胞多胺合成、增殖和侵袭的抑制作用。方法采用活细胞计数法观察Ad-ODC-AdoMetDCas对肺癌A-549细胞生长增殖的影响，采用Western印迹法和高效液相色谱分别检测腺病毒对肺癌A-549细胞中ODC和AdoMetDC蛋白表达以及细胞内多胺含量，采用活细胞计数法和流式细胞仪分别检测腺病毒介导的鸟氨酸脱羧酶反义RNA和Ad-ODC-AdoMetDCas对A-549细胞增殖和细胞周期分布的影响，应用Matrigel侵袭实验分析腺病毒对肺癌细胞侵袭活性的改变。结果当感染效率为50时，腺病毒对A-549细胞的感染率约75％。Ad-ODC-AdoMetDCas可明显抑制A-549细胞ODC和AdoMetDC的表达。A-549细胞感染Ad-ODC-AdoMetDCas后，细胞内多胺含量明显降低。感染Ad-ODC-AdoMetDCas的A-549细胞停滞在G1期。Ad-ODC-AdoMetDCas可显著降低A-549细胞的体外侵袭能力。结论ODC和AdoMetDC双反义腺病毒能够显著抑制肺癌A-549细胞的增殖和侵袭。

Objective To study the inhibitory effects of antisense bicistronic recombinant adenovirus vector of ornithine decarboxylase （ ODC ） and S-adenosylmethionine decarboxylase （ Ad-ODC- AdoMetDCas） on polyamine biosynthesis,proliferation and invasion of lung cancer cells. Methods Adenovirusmediated gene transduction efficiency was assessed with counting GFP-positive cells using trypan blue. Western Blot and HPLC were used to detect ODC and S-AdoMetDC expression and polyamine content in A- 549 cells respectively. Viable cell counting and cell cycle analysis were adopted to evaluate cell growth and cell cycle distribution, and A-549 cell invasion in vitro was detected with Matrigel invasion assay. Results Approximate 75% of A-549 cells were infected with Ad-ODC-AdoMetDCas when multiplicity of infection reached 50. Our study demonstrated that Ad-ODC-AdoMetDCas vector-mediated gene transfer inhibited tumor cell growth through the blockade of polyamine synthesis pathway. The tumor cells were arrested at cell cycle G1 phase after gene transfer. Gene transferred tumor cells were shown to possess markedly decreased invasiveness. Conclusion Ad-ODC-AdoMetDCas has significant inhibitory effects on lung cancer cell proliferation and invasion and bears therapeutic potential for the treatment of lung cancer.