Zhang Hailong, Liu Xiaogang, Wu Fengbo, Qin Feifei, Feng Ping, Xu Ting, Li Xiang, Yang Li
State Key Laboratory of Biotherapy, West China Hospital, West China Medical School, Sichuan University, Chengdu 610041, China.
Department of Gastroenterology, Hospital of the University of Electronic Science and Technology of China and Sichuan Provincial People's Hospital, Chengdu 610041, China.
Int J Mol Sci. 2016 May 17;17(5):676. doi: 10.3390/ijms17050676.
Prostate cancer (PCa) is a malignant tumor for which there are no effective treatment strategies. In this study, we developed a targeted strategy for prostate-specific membrane-antigen (PSMA)-positive PCa in vitro based on 2-(3-((S)-5-amino-1-carboxypentyl)ureido) pentanedioic acid (ACUPA) modified polyethylene glycol (PEG)-Cholesterol micelles containing wogonin (WOG), which was named ACUPA-M-WOG. ACUPA-M-WOG was conventionally prepared using a self-assembling method, which produced stable particle size and ζ potential. Moreover, ACUPA-M-WOG showed good drug encapsulating capacity and drug release profiles. Fluorescence activated cell sorting (FACS) results suggested that ACUPA modified PEG-Cholesterol micelles could effectively enhance the drug uptake on PSMA(+) PCa cells, and the cytotoxicity of ACUPA-M-WOG was stronger than other controls according to in vitro cellular proliferation and apoptosis assays, separately through methyl thiazolyl tetrazolium (MTT) and Annexin V/Propidium Iodide (PI) staining. Finally, the molecular mechanisms of ACUPA-M-WOG's effects on human PSMA(+) PCa were investigated, and were mainly the intrinsic or extrinsic apoptosis signaling pathways. The Western blot results suggested that ACUPA-M-WOG could enhance the WOG-induced apoptosis, which was mainly via the intrinsic signaling pathway rather than the extrinsic signaling pathway. In conclusion, ACUPA-M-WOG was successfully developed for WOG-selective delivery to PSMA(+) PCa cells and had stronger inhibition than free drugs, which might make it an effective strategy for PSMA(+) PCa.
前列腺癌（PCa）是一种尚无有效治疗策略的恶性肿瘤。在本研究中，我们基于2-(3-((S)-5-氨基-1-羧基戊基)脲基)戊二酸（ACUPA）修饰的聚乙二醇（PEG）-胆固醇胶束，开发了一种体外靶向前列腺特异性膜抗原（PSMA）阳性PCa的策略，该胶束含有汉黄芩素（WOG），命名为ACUPA-M-WOG。ACUPA-M-WOG采用自组装方法常规制备，具有稳定的粒径和ζ电位。此外，ACUPA-M-WOG表现出良好的药物包封能力和药物释放曲线。荧光激活细胞分选（FACS）结果表明，ACUPA修饰的PEG-胆固醇胶束可有效增强PSMA(+) PCa细胞对药物的摄取，根据体外细胞增殖和凋亡试验，分别通过甲基噻唑基四氮唑（MTT）和膜联蛋白V/碘化丙啶（PI）染色，ACUPA-M-WOG的细胞毒性比其他对照更强。最后，研究了ACUPA-M-WOG对人PSMA(+) PCa作用的分子机制，主要是内源性或外源性凋亡信号通路。蛋白质印迹结果表明，ACUPA-M-WOG可增强WOG诱导的凋亡，主要通过内源性信号通路而非外源性信号通路。总之，成功开发了ACUPA-M-WOG用于将WOG选择性递送至PSMA(+) PCa细胞，且其抑制作用比游离药物更强，这可能使其成为治疗PSMA(+) PCa的有效策略。
