目的: 探讨抗前列腺癌多肽APP216对体外培养的前列腺癌细胞的杀伤作用，为抗前列腺癌新药的研究奠定基础。方法：利用MTT实验、细胞凋亡染色及流式细胞仪,检测包含有BH3、K237、DG2结构域和能被PSA特异性水解的短肽序列的多肽药物APP216对分泌PSA的前列腺癌细胞系LNCaP、22RV1及不分泌PSA的前列腺癌细胞系PC3m、DU145的杀伤作用。结果：APP216（270 μg/ml）处理48 h后，前列腺癌细胞系LNCaP、22RV1的细胞生存率分别为22%和34%，72 h后为10%和8%；前列腺癌细胞系PC3m、DU145的细胞生存率分别为90%和95%，72 h后为87%和92%。APP216作用后，分泌PSA的前列腺癌细胞胞核呈致密浓染，或呈碎块状，有凋亡小体出现；不分泌PSA的PC3m细胞则未发现改变。APP216（270 μg/ml）处理48 h后，分泌PSA的LNCaP细胞凋亡率为36.26%，不分泌PSA的PC3m细胞凋亡率仅为1.63%。结论：APP216多肽对分泌PSA的前列腺癌细胞有杀伤作用，可诱导肿瘤细胞发生凋亡；而对于不分泌PSA的前列腺癌细胞则效果不佳。证实了该多肽可被PSA特异性酶切；同时，BH3结构域可通过HIV-TAT的转导作用转入细胞内诱导凋亡。

Objective: To investigate the in vitro the anti-prostate cancer effect of a novel polypeptide, APP216, so as to provide a basis for development of new drug for treatment of prostate cancer. Methods: The polypeptide drug included the amino sequences of BH3, K237 and DG2 domain and the peptide that could be digested by PSA. The anti-prostate cancer effects of the polypeptide prodrug on prostate cancer cell line LNCaP, 22RV1 (secreting PSA) and PC3, DU145 (secreting no PSA) were determined by MTT test and Hoechst 33258 staining. Results: MTT test revealed that the surviving rates of LNCaP and 22PV1 cells were respectively 22% and 34% 48 h after APP216 (270 μg/ml) treatment, and 9.8% and 8.2% 72 h after APP216 (270 μg/ml) treatment. The surviving rates of PC3 and DU145 cells were respectively 90% and 95% 48 h after APP216 (270 μg/ml) treatment, and 87% and 92% 72 h after APP216 (270 μg/ml) treatment. Hoechst 33258 staining showed the typical features of cell apoptosis: cell shrinkage, chromatin condensation and hypodiploid genomic DNA content in LNCaP and PC3m cells. Flow cytometry showed an apotosis rate of 36.26% 48 h after APP216 (270 μg/ml) treatment in LNCaP cells, and of 1.63% after 48 h APP216 (270 μg/ml) treatment in PC3m cells.Conclusion: APP216 has a satisfactory in vitro cytotoxicity on human PSA-secreting prostate cancer cells and can induce tumor cell apoptosis, but not on non-PSA secreting prostate cancer cells, indicating APP216 polypeptide can be specifically digested by endonuclease enzyme. Meanwhile, BH3 domain can be transferred into cells and induce apoptosis through HIV-TAT.

国家高新技术研究发展规划(863)资助项目(NO.2001AA215321)