Retrovirus-mediated HSV_1-tk gene therapy for androgen-independent prostate cancer / 中华泌尿外科杂志

ABSTRACT

Objective To use HSV 1 tk (herpes simplex virus type Ⅰthymidine kinase)/GCV (ganciclovir) in androgen independent prostate cancer cells(C4 2,PC3) in vitro in order to provide useful basis for clinical use. Methods HSV 1 tk gene was ligated to a pN 2A retroviral vector. Recombinant DNA molecules being introduced into a packaging cell line PA317,the high titer virus producer cells (VPC) were screened.The integration and expression of HSV 1 tk gene in VPC was observed by PCR and RT PCR. VPC was co cultured with these cancer cells in the light of 1∶1,1∶2,1∶4,1∶8.Cell viability (cytotoxicity) was assessed by SRB (sulforhodamine B protein dye binding)after the first day,the third day,the fifth and the seventh day. Results The highest titer VPC producing HSV 1 tk gene was isolated. Retrovirus mediated HSV 1 tk gene therapy was effective and active against such prostate cancer cells.The best one was co culture of VPC and cancer cells at 1∶1 and the fifth day followed by GCV. Compared with C4 2, PC3 decreased remarkably.The activation of apoptosis and other ways failed to be found. Conclusions Retrovirus mediated HSV 1 tk gene therapy in Vitro directly killed the tumor cells by cytolytic activity.