Shi Sheng-Yu, Zhao Liang-Wei, Liu Chong-Bing, Xiao Hua-Ming, Jiang Zhong-Jun
Department of General Surgery, Huzhou Traditional Chinese Medicine Hospital Affiliated to Zhejiang Chinese Medical University, Huzhou, 313000, Zhejiang, China.
Department of Urology, Huzhou Traditional Chinese Medicine Hospital Affiliated to Zhejiang Chinese Medical University, Huzhou, 313000, Zhejiang, China.
Naunyn Schmiedebergs Arch Pharmacol. 2025 May;398(5):5915-5925. doi: 10.1007/s00210-024-03652-0. Epub 2024 Dec 2.
Breast cancer is a common malignant tumor in women and triple-negative breast cancer (TNBC) is the most challenging type of breast cancer with poor prognosis. We aimed to elucidate the effects of pirfenidone, a FDA-approved oral anti-fibrotic drug which has recently shown antitumor potential, in the progression of TNBC and the underlying mechanisms. After TNBC cells were treated with pirfenidone, cell viability was evaluated using CCK-8 assay. The EDU staining was applied for reflecting the ability of cell proliferation. Additionally, cell cycle distribution and apoptotic rate of TNBC cells exposed to pirfenidone were determined by flow cytometry. The levels of proteins associated with cell cycle, apoptosis and Hedgehog/gliomaassociated oncogene homolog (GLI)1 signaling was examined through western blot. Then, GLI1 was upregulated to analyze the proliferation, cell cycle and apoptosis of TNBC cells in the presence of pirfenidone to reveal the regulatory mechanism. Pirfenidone dose-dependently decreased the viability and proliferation of MDA-MB-231 and HCC-1937 cells. Besides, the distribution of TNBC cells in G0/G1 phase was significantly elevated by pirfenidone, accompanied by downregulated levels of CyclinD1, CDK4 and CDK6. Simultaneously, pirfenidone caused remarkably increased apoptotic MDA-MB-231 and HCC-1937 cells, coupled with downregulated BCL2 expression as well as upregulated Bax, cleaved caspase3 and cleaved PARP expression. Expression of proteins related to Hedgehog/gliomaassociated oncogene homolog (GLI)1 signaling was tested through western blot. Particularly, GLI1 and PTCH1 levels were dose-dependently inhibited after pirfenidone exposure. Interestingly, GLI1 overexpression attenuated the influences of pirfenidone on the proliferation, cell cycle and apoptosis of TNBC cells. Collectively, pirfenidone arrests the cell cycle and promotes apoptosis of TNBC cells by suppressing Hedgehog/GLI1 signaling.
乳腺癌是女性常见的恶性肿瘤，三阴性乳腺癌（TNBC）是最具挑战性的乳腺癌类型，预后较差。我们旨在阐明吡非尼酮（一种最近显示出抗肿瘤潜力的FDA批准的口服抗纤维化药物）对TNBC进展的影响及其潜在机制。用吡非尼酮处理TNBC细胞后，使用CCK-8法评估细胞活力。应用EDU染色来反映细胞增殖能力。此外，通过流式细胞术测定暴露于吡非尼酮的TNBC细胞的细胞周期分布和凋亡率。通过蛋白质印迹法检测与细胞周期、凋亡和刺猬因子/胶质瘤相关癌基因同源物（GLI）1信号通路相关的蛋白质水平。然后，上调GLI1以分析在吡非尼酮存在下TNBC细胞的增殖、细胞周期和凋亡，以揭示调节机制。吡非尼酮剂量依赖性地降低MDA-MB-231和HCC-1937细胞的活力和增殖。此外，吡非尼酮显著提高了TNBC细胞在G0/G1期的分布，同时细胞周期蛋白D1、细胞周期蛋白依赖性激酶4（CDK4）和细胞周期蛋白依赖性激酶6（CDK6）水平下调。同时，吡非尼酮导致MDA-MB-231和HCC-1937细胞凋亡显著增加，同时BCL2表达下调，而Bax、裂解的半胱天冬酶3和裂解的聚（ADP-核糖）聚合酶（PARP）表达上调。通过蛋白质印迹法检测与刺猬因子/胶质瘤相关癌基因同源物（GLI）1信号通路相关的蛋白质表达。特别是，吡非尼酮暴露后，GLI1和PTCH1水平呈剂量依赖性抑制。有趣的是，GLI1过表达减弱了吡非尼酮对TNBC细胞增殖、细胞周期和凋亡的影响。总的来说，吡非尼酮通过抑制刺猬因子/GLI1信号通路来阻滞TNBC细胞的细胞周期并促进其凋亡。
