Dehydrodiisoeugenol Targets the PLK1-p53 Axis to Inhibit Breast Cancer Cell Cycle

Wenbin Huang ^1* Lin Li ¹ Yifan Zheng ¹ Yongxia Yang ² Senlin Shi ¹ Shangjie Liu ¹ Keying Huang ¹ Luonan Qiu ² Rongxin Zhang ³ Yin Leng ^1*

• ¹ The First Affiliated Hospital of Guangdong Pharmaceutical University, Guangzhou, China
• ² Guangdong Cosmetics Engineering &Technology Research Center, Guangdong Pharmaceutical University, Guangzhou, Guangdong Province, China
• ³ Guangdong Provincial Key Laboratory of Biotechnology Candidate Drug Research, School of Life Science and Biopharmaceuticals, Guangdong Pharmaceutical University, Guangzhou, Guangdong Province, China

Alpinia Katsumadai Hayata, a traditional Chinese medicine derived from the nearly mature seed of the nutmeg plant in the ginger family, possesses a range of pharmacological properties, including gastric mucosa protection, anti-ulcer, gastrointestinal function enhancement, antiemetic, antibacterial, antioxidant, anti-inflammatory, and antitumor effects. Despite these benefits, the specific compounds and molecular mechanisms underlying its potential in breast cancer treatment remain elusive. This study utilized the TCMSP database to identify seven bioactive compounds in Alpinia Katsumadai Hayata, with dehydrodiisoeugenol (DHIE) emerging as a promising candidate against breast cancer through LC-MS validation and disease enrichment analysis. Bioinformatics analyses, including network pharmacology, transcriptome difference analysis, single-cell sequencing, and molecular docking, suggest that DHIE inhibits breast cancer cell proliferation by modulating the PLK1-p53 signaling axis. CCK-8 assays confirmed that DHIE suppresses breast cancer cell growth within a specific concentration range without harming mammary epithelial cells. Flow cytometry indicated that DHIE arrests the cell cycle at the G0/G1 phase, while Western blotting revealed its downregulation of PLK1 and differential modulation of wtp53/mtp53 expression in various breast cancer cell types, contributing to cell cycle arrest. In vivo studies and immunohistochemistry further demonstrated that the combination of DHIE and DOX enhances survival rates and inhibits breast cancer progression, as well as reducing Ki-67 positivity. Collectively, these findings suggest that DHIE from Alpinia Katsumadai Hayata exhibits anti-breast cancer activity, potentially through the PLK1-p53 pathway.