Gao H, Song Q, Yang J, Yu S, Zhao J, Yu G
Department of Urology, the Dalian Central Hospital, Dalian, China.
Department of Urology, the Affiliated Hospital of Dalian Medical University, Dalian, China.
Cell Mol Biol (Noisy-le-grand). 2017 Aug 30;63(8):104-108. doi: 10.14715/cmb/2017.63.8.22.
To investigate the effect of carnosol on the Hedgehog (HH) signaling pathway in human hormone-dependent prostate cancer cell line LNCaP and hormone-independent prostate cancer cell line DU145. The expression levels of glioma-associated oncogene homolog 1 (Gli1) and Sonic hedgehog (Shh) in human prostate cancer tissues were detected by immunohistochemistry. After treated with carnosol (0.25-16 μmol/L), the cell survival of LNCaP and DU145 cells were detected by MTT assay. The expression levels of Gli1 and Shh mRNA and protein in the two cells were detected by qRT-PCR and western blot, respectively. The apoptosis was determined by the caspase-3 activity assay. Results showed that Shh and Gli1 were upregulated in cancer tissues. The inhibitory effect of carnosol on cell survival was enhanced with concentration, suggesting both LNCaP and DU145 cells were sensitive to carnosol. The inhibitory effects of carnosol on Gli1 and Shh mRNAs in the hormone-dependent LNCaP prostate cancer cell was stronger than that in the hormone-independent DU145 prostate cancer cells. Carnosol downregulated the expression of Gli1 in nucleus, and Shh in cells. Greater carnosol concentration resulted in lower levels of Gli1 and Shh. Carnosol increased caspase-3 activity in a dose-dependent manner, suggesting that carnosol promotes cell apoptosis. Thus, carnosol can inhibit the proliferation and induce the apoptosis of prostate cancer cells in vitro, and its mechanism might be associated with the inhibiting of HH signaling pathway. Although the inhibitory effect of carnosol on hormone-dependent LNCaP prostate cancer cells is stronger than hormone-independent DU145 prostate cancer cells, carnosol might be a potential drug for hormone-independent prostate cancer.
研究鼠尾草酸对人激素依赖性前列腺癌细胞系LNCaP和激素非依赖性前列腺癌细胞系DU145中刺猬（HH）信号通路的影响。采用免疫组织化学法检测人前列腺癌组织中胶质瘤相关癌基因同源物1（Gli1）和音猬因子（Shh）的表达水平。用鼠尾草酸（0.25 - 16 μmol/L）处理后，采用MTT法检测LNCaP和DU145细胞的存活率。分别用qRT-PCR和蛋白质印迹法检测两种细胞中Gli1和Shh mRNA及蛋白的表达水平。通过caspase-3活性测定法测定细胞凋亡情况。结果显示，癌组织中Shh和Gli1表达上调。鼠尾草酸对细胞存活的抑制作用随浓度增加而增强，提示LNCaP和DU145细胞对鼠尾草酸均敏感。鼠尾草酸对激素依赖性LNCaP前列腺癌细胞中Gli1和Shh mRNA的抑制作用强于激素非依赖性DU145前列腺癌细胞。鼠尾草酸下调细胞核中Gli1的表达以及细胞中Shh的表达。鼠尾草酸浓度越高，Gli1和Shh水平越低。鼠尾草酸以剂量依赖性方式增加caspase-3活性，提示鼠尾草酸促进细胞凋亡。因此，鼠尾草酸在体外可抑制前列腺癌细胞增殖并诱导其凋亡，其机制可能与抑制HH信号通路有关。尽管鼠尾草酸对激素依赖性LNCaP前列腺癌细胞的抑制作用强于激素非依赖性DU145前列腺癌细胞，但鼠尾草酸可能是激素非依赖性前列腺癌的一种潜在药物。
