Yang Haolin, Bi Yu, Wang Min, Liu Yan, Naseem Anam, Huang Jinchang
Beijing University of Chinese Medicine, Beijing, China.
Heilongjiang University of Chinese Medicine 24 Heping Road, Xiangfang District, Harbin 150040, China.
J Ethnopharmacol. 2025 Mar 26;344:119556. doi: 10.1016/j.jep.2025.119556. Epub 2025 Feb 24.
Xihuang pill (XHP), a traditional prescription for treating "Ru yan (breast cancer)" for 270 years, has the efficacies of clearing heat and improving blood circulation. Previous research has demonstrated that XHP yields significant therapeutic effects in the management of triple-negative breast cancer (TNBC).
To explore and validate the usage of a traditional Chinese medicine formula XHP in combating TNBC by epithelial-mesenchymal transition (EMT) and apoptosis mechanisms.
The cytotoxic effects of XHP were evaluated using the CCK-8 assay after incubating with 4T1 and MDA-MB-231 TNBC cell lines. WB assay was conducted to detect the expression of proteins associated with EMT and apoptosis signaling pathways, elucidating the mechanism by which XHP inhibits TNBC. DAPI staining, and mitochondrial membrane potential (MMP) assays (JC-1) were utilized to further confirm the effect of apoptosis in TNBC. Moreover, morphological examination, wound healing assays, and transwell experiments were used to investigate the EMT properties of XHP. Besides, this study constructed 4T1-xenograft model to assess the anti-tumor efficacy of XHP, while the underlying mechanism was revealed by H&E staining.
XHP suppressed tumor growth and metastasis in Balb/c transplanted tumor model by inducing EMT and apoptosis. XHP modulated the expression of the AXL/TGF-β1/RhoA/ROCK pathway, reducing tumor cell EMT, and influencing the ERK signaling pathway. Moreover, XHP inhibited the expression level of Bcl2, Bax and caspase 9 to promote apoptosis in tumor cells. These phenomena eventually manifested as inhibiting the proliferation and migration of TNBC cells.
XHP inhibits TNBC and regulates EMT and apoptosis phenotypes by regulating the AXL/TGF-β1/RhoA/ROCK and ERK pathways. Therefore, it could be further utilized in the discovery of anti-cancer drugs.
西黄丸（XHP）是一种治疗“乳岩（乳腺癌）”已有270年历史的传统方剂，具有清热和活血化瘀的功效。先前的研究表明，西黄丸在三阴性乳腺癌（TNBC）的治疗中产生显著的治疗效果。
通过上皮-间质转化（EMT）和凋亡机制探索并验证中药方剂西黄丸在对抗三阴性乳腺癌中的作用。
将西黄丸与4T1和MDA-MB-231三阴性乳腺癌细胞系孵育后，使用CCK-8法评估其细胞毒性作用。进行蛋白质免疫印迹（WB）分析以检测与EMT和凋亡信号通路相关的蛋白质表达，阐明西黄丸抑制三阴性乳腺癌的机制。利用4',6-二脒基-2-苯基吲哚（DAPI）染色和线粒体膜电位（MMP）检测（JC-1）进一步确认西黄丸对三阴性乳腺癌细胞凋亡的影响。此外，通过形态学检查、伤口愈合试验和Transwell实验研究西黄丸的EMT特性。此外，本研究构建了4T1异种移植模型以评估西黄丸的抗肿瘤疗效，同时通过苏木精-伊红（H&E）染色揭示其潜在机制。
西黄丸通过诱导EMT和凋亡抑制Balb/c移植瘤模型中的肿瘤生长和转移。西黄丸调节AXL/转化生长因子-β1（TGF-β1）/RhoA/ Rho相关卷曲螺旋形成蛋白激酶（ROCK）信号通路的表达，减少肿瘤细胞的EMT，并影响细胞外信号调节激酶（ERK）信号通路。此外，西黄丸抑制B细胞淋巴瘤-2（Bcl2）、Bax和半胱天冬酶9（caspase 9）的表达水平以促进肿瘤细胞凋亡。这些现象最终表现为抑制三阴性乳腺癌细胞的增殖和迁移。
西黄丸通过调节AXL/TGF-β1/RhoA/ROCK和ERK信号通路抑制三阴性乳腺癌并调节EMT和凋亡表型。因此，它可进一步用于抗癌药物的研发。
