Du Guang, Zhang Zheng-wang, Zhang Yong-kang, Guo Jian-ming
Department of Urology, ST. Luke's Hospital, Shanghai 200050, China.
Zhonghua Nan Ke Xue. 2007 Jun;13(6):563-7.
To study the action mechanism of a Chinese herbal mixture PC-SPES II inducing the apoptosis of androgen independent prostate adenocarcinoma cell line (PC-3).
The growth of PC-3 was shown by MTT. Immunofluorescence staining of acridine orange (AO) and flow cytometry were used to detect the apoptosis. The expressions of the apoptosis-related proteins were analyzed with their monoclonal or polyclonal antibodies after Western blotting.
PC-SPES II not only inhibited the growth of PC-3 cells but also induced their death. The apoptosis of PC-3 cells treated with PC-SPES II was detected by immunofluorescence staining of AO and flow cytometry, which showed the apoptotic cells to be (29.8 +/- 5.6)%, but the untreated control cells (0.06 +/- 0.014)%, (P < 0.01). The expression of Bcl-2 and Bcl-xL, two antiapoptosis proteins, was decreased while Bax, a pro apoptosis protein, was elevated in the cells treated with PC-SPES II as compared with the untreated control (P < 0.01). Accordingly, the expression of the activated fragments of caspase-3 was also increased (P < 0.01).
The Chinese herbal mixture PCSPES II can induce the apoptosis of PC-3. Its mechanism may lie in the up-regulation of Bax expression and down-regulation of Bcl-2 and Bcl-xL expressions, which induce the activated fragments of caspase-3 by mitochondria.
研究中药复方PC-SPES II诱导雄激素非依赖性前列腺腺癌细胞系(PC-3)凋亡的作用机制。
采用MTT法检测PC-3细胞的生长情况。用吖啶橙(AO)免疫荧光染色和流式细胞术检测细胞凋亡。Western印迹后用单克隆或多克隆抗体分析凋亡相关蛋白的表达。
PC-SPES II不仅抑制PC-3细胞的生长,还诱导其死亡。通过AO免疫荧光染色和流式细胞术检测PC-SPES II处理的PC-3细胞凋亡情况,结果显示凋亡细胞为(29.8±5.6)%,而未处理的对照细胞为(0.06±0.014)%,(P<0.01)。与未处理的对照相比,PC-SPES II处理的细胞中两种抗凋亡蛋白Bcl-2和Bcl-xL的表达降低,而促凋亡蛋白Bax的表达升高(P<0.01)。相应地,caspase-3活化片段的表达也增加(P<0.01)。
中药复方PCSPES II可诱导PC-3细胞凋亡。其机制可能在于上调Bax表达,下调Bcl-2和Bcl-xL表达,通过线粒体诱导caspase-3活化片段的产生。