腺病毒介导的凋亡素、内皮抑素双基因对食管癌的实验研究

目的探讨携带有凋亡素(vp3、Apoptin)、内皮抑素(Endostatin)双基因的重组腺病毒载体在食管癌等多种肿瘤细胞内的表达情况及致凋亡作用,为进一步的食管癌治疗应用研究打下基础。方法将已纯化的携带凋亡素和内皮抑素基因的腺病毒Ad-vp3-IRES-sEndo-His感染食管癌细胞Eca-109、小鼠肝癌细胞Hepa1-6、结肠癌细胞LoVo提取各种细胞RNA,针对凋亡素、内皮抑素基因的表达进行RT-PCR检测,同时对感染腺病毒Ad-vp3-IRES-sEndo-His、Ad-vp3的食管癌细胞通过流式细胞仪、Hoechst33258染色等方式进行细胞凋亡率的检测。结果 RT-PCR检测显示被感染的多种肿瘤细胞均可表达凋亡素、内皮抑素基因的mRNA,Ho-echst33258染色显示被Ad-vp3-IRES-sEndo-His感染的食管癌细胞出现凋亡形态学改变,用流式细胞仪测定时段最高凋亡率达47.7%,与对照组相比具有统计学意义(P<0.05)。结论携带凋亡素及内皮抑素双基因的腺病毒载体能在多种肿瘤细胞中表达,感染食管癌细胞后,能有效诱导其凋亡且其致凋亡率高于仅携带凋亡素的单基因腺病毒载体。

The effect of adenovirus vector carrying apoptin and endostatin genes on the esophageal cancer

Objective To investigate the effects and expression of the recombinant adenovirus vector that carrying double genes(apoptin and endostatin) in esophageal cancer cells and to find new therapy for the treatment of esophageal cancer.Methods After Ad-vp3-IRES-sEndo-His that carrying apoptin and endostatin genes infected the human esophageal cancer cell Eca-109,mouse cancer cell Hepa1-6,colon cancer cell LoVo,the expression of apoptin and endoststion genes were detected by RT-PCR,and the apoptosis of Eca-109 cell was detected by fluorescent staining and flow cytometer(FCM).Results RT-PCR assay detected that these cells could express apoptin and endostatin mRNA,and the apoptosis in Eca-109 esophageal cancer cells cell after infected by Ad-vp3-IRES-sEndo-His was demonstrated by stain with Hoechst33258.The rate of apoptosis in Eca-109 cell(47.7%) was higher than that of the control group that infected by ad-VP3(P<0.05).Conclusion The Ad-vp3-IRES-sEndo-His that carring the double genes can be expressed in different cancer cells,and it can induce effective apoptosis in the Eca-109 cell and the rate of apoptosis is higher than that induced by Ad-vp3.