{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T12:13:27Z","timestamp":1785413607444,"version":"3.56.0"},"reference-count":68,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2026,4,22]],"date-time":"2026-04-22T00:00:00Z","timestamp":1776816000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["82360720"],"award-info":[{"award-number":["82360720"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"award":["82360720"],"award-info":[{"award-number":["82360720"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pharmaceuticals"],"abstract":"<jats:p>Background: Prostate cancer (PCa) is the leading male urinary malignancy globally. Our previous article demonstrated the anti-PCa activity of Euphorbia humifusa Willd water extract (EHW) and some of its compounds via downregulating AR expression, but the anti-PCa active compounds from Euphorbia humifusa Willd (EH) and their mechanisms of action are yet to be clarified. Thus, the current article studied the in vitro anti-PCa effects of 3,3\u2032-di-O-methylellagic acid (3,3\u2032-di-O-Me-EA) derived from EHW and the related mechanism involved. Methods: 3,3\u2019-di-O-Me-EA was isolated from EHW applying bioassay-guided fractionation. The spectroscopic methods were used to determining the structure of 3,3\u2032-di-O-Me-EA. The drug-likeness and ADMET properties (absorption, distribution, metabolism, excretion, and toxicity) of 3,3\u2032-di-O-Me-EA were analyzed in silico. Molecular docking and real-time surface plasmon resonance (SPR) analysis were performed to measure the interaction of 3,3\u2032-di-O-Me-EA and VDAC1 protein. The viability and apoptosis of 22RV-1 and DU145 PCa cells were determined using MTT and Annexin V-FITC staining assay, respectively. q-PCR and Western blot experiments were used to analyzing the gene and protein expressions of VDAC1. Results: 3,3\u2032-di-O-Me-EA was isolated and purified from EHW with a purity of \u226590.06%, and its structure was identified by HRTOF mass, NMR, and an authentic standard. In silico ADMET analysis indicated its favorable drug-like and pharmacokinetic properties. Molecular docking and SPR results confirmed that 3,3\u2032-di-O-Me-EA could bind with the VDAC1 protein. Moreover, 3,3\u2032-di-O-Me-EA dose- and time-dependently inhibited 22RV-1 and DU145 PCa cell viability, and induced apoptosis in a dose-dependent manner (p &lt; 0.05). RT-qPCR and Western blot results showed that 3,3\u2032-di-O-Me-EA dose-dependently up-regulated VDAC1 gene and protein expression levels in 22RV-1 and DU145 cells (p &lt; 0.05). Meanwhile, in VDAC1-depleted 22RV-1 and DU145 cells, 3,3\u2032-di-O-Me-EA down-regulated VDAC1 gene and protein expression levels, increased cell viability, and inhibited apoptosis compared to 22RV-1 and DU145 cells (p &lt; 0.05). Furthermore, 3,3\u2032-di-O-Me-EA enhanced VDAC1 gene and protein expression levels, inhibited cell viability, and induced apoptosis in VDAC1-overexpressed 22RV-1 and DU145 cells compared with 22RV-1 and DU145 cells (p &lt; 0.05). Overall, EH active compound 3,3\u2032-di-O-Me-EA may inhibit viability and induce apoptosis of 22RV-1 and DU145 PCa cells via up-regulating VDAC1 gene and protein expression levels. Conclusion: The results indicated that the 22RV1 and DU145 PCa cell viability inhibitory effects of 3,3\u2032-di-O-Me-EA isolated from EH may be mediated by induction of apoptosis through up-regulation of VDAC1 gene and protein expression levels.<\/jats:p>","DOI":"10.3390\/ph19050652","type":"journal-article","created":{"date-parts":[[2026,4,22]],"date-time":"2026-04-22T12:00:32Z","timestamp":1776859232000},"page":"652","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["3,3\u2032-Di-O-methylellagic Acid Isolated from Euphorbia humifusa Willd Suppresses Prostate Cancer Cell Viability via Regulating VDAC1 Protein Expression"],"prefix":"10.3390","volume":"19","author":[{"given":"Gulijikere","family":"Kuerban","sequence":"first","affiliation":[{"name":"College of Pharmacy, Xinjiang Medical University, Shang De North Road, Shui Mo Gou District, Urumqi 830017, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xinling","family":"Wang","sequence":"additional","affiliation":[{"name":"College of Pharmacy, Xinjiang Medical University, Shang De North Road, Shui Mo Gou District, Urumqi 830017, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chengjing","family":"Shen","sequence":"additional","affiliation":[{"name":"College of Pharmacy, Xinjiang Medical University, Shang De North Road, Shui Mo Gou District, Urumqi 830017, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mireguli","family":"Abulimiti","sequence":"additional","affiliation":[{"name":"College of Pharmacy, Xinjiang Medical University, Shang De North Road, Shui Mo Gou District, Urumqi 830017, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jialu","family":"Hu","sequence":"additional","affiliation":[{"name":"College of Pharmacy, Xinjiang Medical University, Shang De North Road, Shui Mo Gou District, Urumqi 830017, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zilala","family":"Yalihong","sequence":"additional","affiliation":[{"name":"College of Pharmacy, Xinjiang Medical University, Shang De North Road, Shui Mo Gou District, Urumqi 830017, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aikebaier","family":"Maimaiti","sequence":"additional","affiliation":[{"name":"College of Pharmacy, Xinjiang Medical University, Shang De North Road, Shui Mo Gou District, Urumqi 830017, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,4,22]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Sekhoacha, M., Riet, K., Motloung, P., Gumenku, L., Adegoke, A., and Mashele, S. 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