摘要
目的:研究携带小鼠干扰素γ(mIFN-γ)基因的溶瘤病毒CNHK300-mIFN-γ(CNHK300-Mγ)对恶性肿瘤细胞的体外杀伤作用。方法:CNHK300-Mγ、CNHK300、ONYX-015和AdEasy-mIFN-γ(AdEasy-Mγ)感染肺癌细胞株A549、肝癌细胞株SMMC-7721、胰腺癌细胞株PANC-1和正常成纤维细胞株BJ,通过病毒增殖实验、细胞病理效应和细胞杀伤实验观察CNHK300-Mγ、CNHK300和ONYX-015在上述肿瘤细胞和正常细胞中的增殖能力和对这些细胞的杀伤作用,通过双抗体夹心ELISA检测CNHK300-Mγ和AdEasy-Mγ在上述肿瘤细胞和正常细胞中不同时相的mIFN-γ表达量。结果:CNHK300-Mγ具有类似于CNHK300的肿瘤细胞选择增殖性,能在恶性肿瘤细胞中大量增殖并杀灭肿瘤细胞,均强于ONYX-015(P<0.01),而在正常细胞中增殖和杀伤作用均较弱,弱于ONYX-015(P<0.05)。CNHK300-Mγ感染恶性肿瘤细胞后有大量的mIFN-γ表达,并随着感染时间延长表达量也相应上升,而AdEasy-Mγ感染后只有少量mIFN-γ的表达,而且感染的时间延长表达量变化也不大(P<0.01)。结论:CNHK300-Mγ能选择性地在肿瘤细胞中增殖,并杀灭肿瘤细胞,同时大量表达具有抗肿瘤作用的蛋白,发挥多重抗瘤作用,有良好的临床应用前景。
AIM： To investigate the cytotoxicity and mouse IFN -γ （mIFN -γ） expression of oncolytic ade- novirus CNHK300 - mIFN - γ （ CNHK300 - Mγ） containing mlFN - y gene in malignant tumor Cells in vitro. METHODS： Human lung cancer cell line A549, human liver cancer cell line SMMC -7721, human pancreatic cancer cell line PANC - 1, and human normal fibroblast line BJ were cultured and treated with CNHK300 - M3γ, CNHK300, ONYX - 015 or AdEasy - mIFN - γ/（ AdEasy - Mγ）. TCID50 assay was used to evaluate the replication ability of CNHK300 - Mγ/ CNHK300 and ONYX-015 in carcinoma cell lines and normal cell line, and the cytotoxicity was evaluated by cytopathic effect assay and MTF assay. The mlFN - γ/expression in the supematant was detected by ELISA after CNHK300 - Mγ/or AdEasy - Mγ/infection in carcinoma cell lines and normal cell line. RESULTS： The tumor - specific replication ability and cytotoxicity of CNHK300 - Mγ were similar to those of CNHK300. The IC50 was as low as MOI of 0.47 pfu/cell for A549 cells, 0. 074 pfu/cell for SMMC - 7721 cells, 0.532 pfu/cell for PANC - 1 cells and was as high as MOI of 281.73 pfu/cell for BJ cells. CNHK300 - Mγ was a more powerful killer of malignant tumor cells than ONYX -015 （P 〈0. 01 ）. The tumor cells infected with CNHK300 - Mγ efficiently expressed mIFN - γ in vitro and mIFN - γ largely increased as the time prolonged in A549, SMMC -7721 and PANC -1 cells. The mIFN -γ expression in the carcinoma cell lines infected with CNHK300 - M3＇ was much higher than that in the cells infected with AdEasy - Mγ, （ P 〈 0.01 ）, but was similar to that in the normal cell line （P 〉 0. 05 ）. CONCLUSION ： CNHK300 - Mγ, selectively replicates and effectively promotes the expression of mIFN γ in carcinoma cells, and specifically kills the tumor cells.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2012年第5期802-806,共5页
Chinese Journal of Pathophysiology
基金
全国优秀博士论文专项基金资助项目(No.FANEDD 200774)
国家自然科学基金国际合作重大项目(No.03102160823)
关键词
溶瘤腺病毒
基因疗法
干扰素γ
肿瘤
Oncolytic adenovirus
Gene therapy
Interferon γ
Neoplasms
