Yu De Bin, Zhong Su Yang, Yang Min, Wang Yi Gang, Qian Qi Jun, Zheng Shu, Liu Xin Yuan
Xinyuan Institute of Medicine and Biotechnology, Zhejiang Sci-Tech University, Hangzhou, China.
Cancer Sci. 2009 Apr;100(4):678-83. doi: 10.1111/j.1349-7006.2009.01110.x. Epub 2009 Mar 1.
Following targeted gene virotherapy, the suppression of tumorigenicity 13 (ST13) gene was inserted into the double-regulated oncolytic adenovirus SG500 to ensure more safety and potent antitumor activity against colorectal cancer in vitro and in vivo. We generated the ST13-expressing oncolytic adenovirus SG500-ST13, with which colorectal carcinoma cell lines SW620 and HCT116, and the lung fibroblast cell line WI38, were infected. Crystal violet staining was carried out to detect the cytopathic effect in cells, and the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method was used to assay cell viability. The effect of apoptosis induced by SG500-ST13 was confirmed by Hoechst staining and the TdT-mediated dUTP-biotin nick-end labeling method. To further identify the antitumor effects of SG500-ST13 on HCT116 xenografts in Balb/c nude mice, the induction of cell death was assessed by hematoxylin-eosin staining. Immunohistochemical study was also carried out.
在进行靶向基因病毒疗法后，将抑癌基因13（ST13）插入双调控溶瘤腺病毒SG500中，以确保其在体外和体内对结直肠癌具有更高的安全性和更强的抗肿瘤活性。我们构建了表达ST13的溶瘤腺病毒SG500-ST13，并使用其感染结直肠癌细胞系SW620和HCT116以及肺成纤维细胞系WI38。通过结晶紫染色检测细胞病变效应，采用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐法检测细胞活力。通过Hoechst染色和TdT介导的dUTP生物素缺口末端标记法证实了SG500-ST13诱导细胞凋亡的作用。为了进一步确定SG500-ST13对Balb/c裸鼠体内HCT116异种移植瘤的抗肿瘤作用，通过苏木精-伊红染色评估细胞死亡的诱导情况。还进行了免疫组织化学研究。
