溶瘤腺病毒和Ad-Endo体外联合作用于宫颈癌细胞的初步研究

In vitro study on combination of oncolytic adenovirus and Ad -Endo against cervical carcinoma

目的探讨溶瘤腺病毒d／1520在宫颈癌细胞（C33A、HeLa）中的复制及其联合Ad—Endo在体外对宫颈癌细胞的杀伤作用；研究dl1520对Ad—Endo在宫颈癌细胞中表达内皮抑素的影响。方法用不同感染复数（MOI）的dl1520和Ad—Endo感染宫颈癌细胞，观察细胞病变效应；用MTT法测定dl1520和Ad—Endo对宫颈癌细胞生长的抑制作用；用ELISA方法检测细胞培养上清中内皮抑素的浓度。结果d／1520导致宫颈癌细胞产生细胞病变；MTT法测定结果显示，Ad—Endo单独作用对宫颈癌细胞的抑制作用很低，d／1520能有效抑制宫颈癌细胞增殖，而且Ad—Endo联合dl1520能产生强于dl1520的抑制效果；Ad—Endo能在宫颈癌细胞中有效表达内皮抑素蛋白，且d／1520联合Ad—Endo组上清中内皮抑素浓度远远高于Ad—Endo单独感染组。结论（1）dl1520可在宫颈癌细胞中复制并杀伤肿瘤细胞；（2）Ad—Endo能促进dl1520对宫颈癌细胞的杀伤作用；（3）dl1520能极大地促进Ad—Endo介导的内皮抑素在宫颈癌细胞中的表达。

Objective To evaluate the anti-cervical carcinoma cells effects of oncolytic adenovirus dl1520 and Ad-Endo in vitro, and investigate the influence of dl1520 on the Endostatin expression directed by Ad-Endo in cervical carcinoma cells. Methods After cervical carcinoma cells were infected with dl1520 or/and Ad-Endo, the cytopathic effects （CPE） were observed, and the cytotoxicities were measured by MTT assay. Endostatin concentrations were measured by ELISA. Results dl1520 can replicate in cervical carcinoma cells and induce cytopathic effects; In MTT assay, Ad-Endo hardly inhibited the growth of cervical carcinoma cells, while dl1520 inhibited efficiently the proliferation of cervical cancer cells. And the inhibitory rates in Ad-Endo ＋ dl1520 group were higher than that in dl1520 group. Ad-Endo expressed Endostatin protein in cervical cancer cells, while Endostatin concentrations in Ad-Endo ＋ dl1520 group were much higer than those in Ad-Endo group. Conclusions dl1520 can kill cervical carcinoma cells in vitro, and this cytotoxicity can be enhanced by combining with Ad-Endo. The expression of Endostatin directed by Ad-Endo in cervical cancer cells can be dramatically increased by dl1520.