Lin Chung-Ming, Chao Min-Chih, Chen Hsin-Han, Chen Hui-Jye
Department of Biotechnology, Ming Chuan University, Taoyuan 33348, Taiwan.
Graduate Institute of Biomedical Sciences, China Medical University, Taichung 40402, Taiwan.
Int J Mol Sci. 2025 Jun 27;26(13):6210. doi: 10.3390/ijms26136210.
Colorectal cancer remains a leading malignancy. As the aberrant activation of Wnt/β-catenin signaling causes colorectal cancer, Wnt/β-catenin signaling inhibitors are potential candidates for colorectal cancer treatment. Our drug screening platform identified ursolic acid (UA), a triterpenoid with various biological activities, as a potential anticancer drug because it inhibits the T-cell factor (TCF)/β-catenin-mediated transcriptional activity. Here, we discovered that UA inhibited Wnt signaling by reducing the Wnt reporter activity and Wnt target gene expression, leading to a delay in cell cycle progression and the suppression of cell proliferation. Stepwise epistatic analyses suggested that UA functions on β-catenin protein stability in Wnt signaling. Further studies revealed that UA reduced β-catenin protein levels by Western blotting and immunofluorescent staining and induced autophagy by microtubule-associated protein 1 light chain 3 beta (LC3B) punctate staining. The cotreatment with UA and the autophagy inhibitors chloroquine and wortmannin recovered the β-catenin protein levels. Therefore, UA was confirmed to induce β-catenin degradation by the autophagy-lysosomal degradation system through inhibition in the phosphatidylinositol 3-kinase (PI3K)/Ak strain transforming (protein kinase B; AKT)/mammalian target of rapamycin (mTOR) signaling pathway. Our results not only highlight the potential of UA in Wnt-driven colorectal cancer therapy but also provide a workable Wnt signaling termination approach for the treatment of other Wnt-related diseases.
结直肠癌仍然是一种主要的恶性肿瘤。由于Wnt/β-连环蛋白信号通路的异常激活会导致结直肠癌,因此Wnt/β-连环蛋白信号通路抑制剂是结直肠癌治疗的潜在候选药物。我们的药物筛选平台鉴定出熊果酸(UA),一种具有多种生物活性的三萜类化合物,作为一种潜在的抗癌药物,因为它能抑制T细胞因子(TCF)/β-连环蛋白介导的转录活性。在此,我们发现UA通过降低Wnt报告基因活性和Wnt靶基因表达来抑制Wnt信号通路,导致细胞周期进程延迟和细胞增殖受到抑制。逐步上位分析表明,UA在Wnt信号通路中对β-连环蛋白的蛋白质稳定性起作用。进一步的研究表明,通过蛋白质免疫印迹法和免疫荧光染色,UA降低了β-连环蛋白的蛋白质水平,并通过微管相关蛋白1轻链3β(LC3B)点状染色诱导自噬。UA与自噬抑制剂氯喹和渥曼青霉素联合处理可恢复β-连环蛋白的蛋白质水平。因此,证实UA通过抑制磷脂酰肌醇3激酶(PI3K)/Akt(蛋白激酶B;AKT)/雷帕霉素哺乳动物靶蛋白(mTOR)信号通路,通过自噬-溶酶体降解系统诱导β-连环蛋白降解。我们的研究结果不仅突出了UA在Wnt驱动的结直肠癌治疗中的潜力,而且为治疗其他Wnt相关疾病提供了一种可行的Wnt信号通路终止方法。