携带小鼠白细胞介素-12基因的增殖型腺病毒对胃癌细胞的作用

Gene therapy for gastric cancer with replication competent adenovirus carrying mouse Interleukin-12 gene

目的观察携带小鼠白细胞介素12(mIL12)基因的增殖型腺病毒载体对胃癌细胞的杀伤作用及mIL12表达情况。方法以携带目的基因的增殖型腺病毒CNHK200mIL12感染人胃癌细胞株SGC7901、人正常肝细胞株L02。首先,通过荧光显微镜下观察、细胞杀伤实验等观察病毒对细胞的杀伤能力;其次,通过Westernblot分析及双抗体夹心法(ELISA)检测mIL12表达情况。结果CNHK200mIL12感染SGC7901后,当MOI=10时即可杀伤大部分的肿瘤细胞,与ONYX015相当,而对正常细胞无明显杀伤。ELISA检测表明mIL12基因表达量达(267.2±34.6)ng/5×105个细胞/48h,较复制缺陷型腺病毒载体高近百倍。结论CNHK200mIL12能在胃癌细胞中复制及增殖杀死胃癌细胞,并高水平表达目的基因。

Objective To investigate the anti tumor effects of replication competent adenovirus carrying mouse interleukin 12 (mIL 12) gene on gastric cancer and the expression level of mIL 12.Methods Replication competent adenovirus,CNHK200 mIL 12,carrying mIL 12 gene was constructed to infect gastric carcinoma cell line SGC 7901 and hepatocyte line L02.The anti tumor potential of CNHK200 mIL 12 to SGC 7901 and L02 was evaluated by fluorescence microscopy and cytotoxicity assay.The expression level of mIL 12 gene was also detected by Western blot and ELISA.Results The vector system CNHK200 mIL 12 possessed the same characteristics as the replication competent virus ONYX 015,replicating and proliferating in the gastric carcinoma cells SGC 7901 but not in the normal cells L02,and could specifically kill most of SGC 7901 at the MOI value of 10,which was 1?000 times more than that by the traditional adenovirus vector.The mIL 12 expression level was also increased,reaching (267.2±34.6) ng/5×10 5 cells/48 h,nearly 1?000 folds more than that by the traditional adenovirus vector carrying mIL 12 gene.Conclusion CNHK200 IL 12 can replicate and proliferate in gastric carcinoma cells,and kill them,meanwhile increase its target gene expression.