Anti-tumor effects of a recombinant anti-prostate specific membrane antigen immunotoxin against prostate cancer cells

BMC Urology

Research Article

Wei-hong Wen¹  Guang-guo Tan²  Qing-chuan Dong³  He Wang⁴  Yan-zhu Wang⁵  Jian-lin Yuan⁵  Ping Meng⁵  Yu-ming Jing⁵  Geng Zhang⁵  Wei-jun Qin⁵  Chen Wang⁶ 

[1] Department of Immunology, Fourth Military Medical University, Xi’an, Shaanxi, China;Department of Pharmaceutical Analysis, School of Pharmacy, Fourth Military Medical University, Xi’an, Shaanxi, China;Department of Urology Surgery, Peoples’ Hospital of Shaanxi Province, Xi’an, Shaanxi, China;Department of Urology, Tangdu Hospital, The Fourth Military Medical University, Xi’an, Shaanxi, China;Department of Urology, Xijing Hospital, Fourth Military Medical University, Xi’an, Shaanxi, China;State Key Laboratory of NBC Protection for Civilian, Beijing, China;

关键词: Gene therapy;    Prostate cancer;    Prostate-specific membrane antigen;    Recombinant protein;    Apoptosis;   

DOI  :  10.1186/s12894-017-0203-9

 received in 2016-09-21, accepted in 2017-02-06,  发布年份 2017

来源: Springer

【 摘 要 】

BackgroundTo evaluate anti-prostate cancer effects of a chimeric tumor-targeted killer protein.MethodsWe established a novel fusion gene, immunocasp-3, composed of NH2-terminal leader sequence fused with an anti-prostate-specific membrane antigen (PSMA) antibody (J591), the furin cleavage sequences of diphtheria toxin (Fdt), and the reverse coding sequences of the large and small subunits of caspase-3 (revcaspase-3). The expressing level of the immunocasp-3 gene was evaluated by using the reverse transcription-PCR (RT-PCR) and western blot analysis. Cell viability assay and cytotoxicity assay were used to evaluate its anti-tumor effects in vitro. Apoptosis was confirmed by electron microscopy and Annexin V-FITC staining. The antitumor effects of immunocasp-3 were assessed in nude mice xenograft models containing PSMA-overexpressing LNCaP cells.ResultsThis study shows that the immunocasp-3 proteins selectively recognized and induced apoptotic death in PSMA-overexpressing LNCaP cells in vitro, where apoptotic cells were present in 15.3% of the cells transfected with the immunocasp-3 expression vector at 48 h after the transfection, in contrast to 5.5% in the control cells. Moreover, LNCaP cells were significantly killed under the condition of the co-culture of the immunocasp-3-secreting Jurkat cells and more than 50% of the LNCaP cells died when the two cell lines were co-cultured within 5 days. In addition, The expression of immunocasp-3 also significantly suppressed tumor growth and greatly prolonged the animal survival rate in vivo.ConclusionA novel fusion gene, immunocasp-3, may represent a viable approach to treating PSMA-positive prostate cancer.

【 授权许可】

CC BY   
© The Author(s). 2017