ODC和AdoMetDC双反义腺病毒载体对食管癌细胞凋亡的影响

Apoptotic effects of ODC and AdoMetDC bi-antisense virus on esophageal cancer cell Eca109

目的探讨ODC和AdoMetDC双反义腺病毒载体(Ad-ODC-AdoMetDCas)对食管癌Eca109细胞凋亡作用的影响。方法应用MTT法实验观察Ad-ODC-AdoMetDCas对食管癌Eca109细胞生长增殖的影响;采用Western印迹和HPLC分别检测腺病毒载体对食管癌Eca109细胞中ODC和AdoMetDC蛋白表达以及胞内多胺含量的抑制作用,同时应用原位末端标记(TUNEL)法观察Ad-ODC-AdoMetDCas对食管癌Eca109细胞凋亡作用的影响,应用透射电镜进一步观察细胞超微结构的改变。结果应用MTT法实验表明,Ad-ODC-AdoMetDCas对食管癌Eca109细胞生长增殖有显著抑制作用(P<0.05)。以Ad-ODC-AdoMetDCas感染食管癌Eca109细胞,可明显抑制食管癌Eca109细胞中ODC和AdoMetDC基因表达(P<0.05)。HPLC结果显示,食管癌Eca109细胞感染Ad-ODC-AdoMetDCas后,细胞内三种多胺含量均明显降低(P<0.05)。TUNEL标记检测结果显示,Ad-ODC-AdoMetDCas可明显引起食管癌Eca109细胞凋亡(P<0.05),透射电镜下可见典型的细胞凋亡特征,表现为细胞体积缩小,核皱缩、碎裂,染色质呈块状边集等。结论ODC和AdoMetDC双反义腺病毒载体能显著抑制食管癌细胞生长增殖,降低细胞多胺合成,促进细胞凋亡,为探讨食管癌基因治疗的可行性提供了实验依据。

Objective To explore the inhibitory effects of Ad-ODC-AdoMetDCas on polyamine biosynthesis and esophageal cancer cell apoptosis. Methods Adenovirus-mediated gene transduction efficiency was assessed by counting the GFP-positive eels by using MTT. The malignant phenotype of the Eca109 cells was assessed by a growth curve. Western Blot and HPLC were used to determine the ODC and AdoMetDC expressions and polyamine content in Eca109 cells. TUNEL was used to analyze cell apoptosis. Morphology of the apoptotic cells was observed by an electron microscope. Result Approximately 70% of the Eca109 cells were infected with Ad-ODC-AdoMetDCas when MOI reached 50. The expression of ODC was inhibited in the infected tumor cells. Ad-ODC- AdoMetDCas could inhibit the Eca109 cell growth and invasive ability. TUNEL proved that Ad-ODC-AdoMetDCas could lead to cell apoptosis. Under an electron microscope, chromatin condensation, nuclear disintegration and formation of apoptotic bodies were found. Conclusion Ad-ODC-AdoMetDCas has significant inhibitory effects on esophageal cancer cell proliferation, leads to celt apoptosis and has therapeutic potential for the treatment of esophageal cancer.