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BMC Urol. 2017 Feb 13;17(1):14. doi: 10.1186/s12894-017-0203-9.

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

Author information

1
Department of Urology, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi, China.
2
Department of Urology Surgery, Peoples' Hospital of Shaanxi Province, Xi'an, Shaanxi, China.
3
Department of Pharmaceutical Analysis, School of Pharmacy, Fourth Military Medical University, Xi'an, Shaanxi, China.
4
Department of Immunology, Fourth Military Medical University, Xi'an, Shaanxi, China.
5
Department of Urology, Tangdu Hospital, The Fourth Military Medical University, Xi'an, Shaanxi, China.
6
State Key Laboratory of NBC Protection for Civilian, Beijing, China.
7
Department of Urology, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi, China. weijunqfmmu@163.com.
8
Department of Urology, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi, China. jianliny@fmmu.edu.cn.

Abstract

BACKGROUND:

To evaluate anti-prostate cancer effects of a chimeric tumor-targeted killer protein.

METHODS:

We 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.

RESULTS:

This 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.

CONCLUSION:

A novel fusion gene, immunocasp-3, may represent a viable approach to treating PSMA-positive prostate cancer.

KEYWORDS:

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

PMID:
28193277
PMCID:
PMC5307788
DOI:
10.1186/s12894-017-0203-9
[Indexed for MEDLINE]
Free PMC Article

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