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Prostate-specific membrane antigen (PSMA), which is highly expressed on prostate cancer cells, represents a promising target for precision drug delivery. In this study, we developed a folate-modified, PSMA-targeting nanoliposome loaded with docetaxel (DFL) to enhance tumor specificity and therapeutic efficacy. Methods: DFL was prepared using a thin-film hydration\u2013sonication method and characterized through physicochemical analyses. Cellular uptake and cytotoxicity were evaluated in PSMA-high LNCaP cells, with PSMA knockdown used to assess target-dependent internalization. Antitumor efficacy was examined with a microfluidic system and LNCaP xenograft nude mice, and safety was evaluated by measuring hepatic and renal biomarkers and performing histopathological analysis of major organs. Results: DFL demonstrated favorable physicochemical properties and significantly enhanced cellular uptake and cytotoxicity in LNCaP cells relative to control formulations. PSMA knockdown markedly attenuated cellular sensitivity to DFL, confirming PSMA-dependent internalization. A 3D microfluidic perfusion platform further corroborated robust and selective DFL uptake under dynamic flow conditions, thereby strengthening the translational relevance of the targeting effect beyond static cultures. In vivo, DFL substantially inhibited tumor progression in LNCaP xenograft models, reducing both tumor volume and weight by more than 50%. TUNEL assays showed increased apoptosis, and immunohistochemistry revealed reduced Ki-67 expression with concomitant upregulation of Caspase-3. No significant alterations in hepatic or renal biomarkers were observed, and histopathological evaluation demonstrated no treatment-associated lesions in major organs. Conclusions: A folate-modified, PSMA-targeting docetaxel nanoliposome was successfully developed, demonstrating enhanced tumor-specific drug delivery and improved antitumor activity with favorable biocompatibility in preclinical models. DFL represents a promising nanomedicine strategy for the precision chemotherapy of prostate cancer.<\/jats:p>","DOI":"10.3390\/pharmaceutics18020244","type":"journal-article","created":{"date-parts":[[2026,2,16]],"date-time":"2026-02-16T11:11:28Z","timestamp":1771240288000},"page":"244","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Harnessing Folate-Mediated PSMA Targeting for Precision Therapy: An Intelligent Liposomal Nanoplatform Against Prostate Cancer"],"prefix":"10.3390","volume":"18","author":[{"given":"Youlong","family":"Hai","sequence":"first","affiliation":[{"name":"Department of Urology, Shanghai Jiao Tong University School of Medicine Affiliated Shanghai Sixth People\u2019s Hospital, 600 Yi Shan Road, Shanghai 200233, China"},{"name":"Department of Hepatobiliary Surgery, Shanghai Pudong Hospital, Fudan University Pudong Medical Center, Shanghai 201399, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jiayi","family":"Ma","sequence":"additional","affiliation":[{"name":"Department of Urology, Shanghai Jiao Tong University School of Medicine Affiliated Shanghai Sixth People\u2019s Hospital, 600 Yi Shan Road, Shanghai 200233, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xuehao","family":"Yu","sequence":"additional","affiliation":[{"name":"Department of Radiology, Shanghai Jiao Tong University School of Medicine Affiliated Shanghai Sixth People\u2019s Hospital, 600 Yi Shan Road, Shanghai 200233, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kun","family":"Zheng","sequence":"additional","affiliation":[{"name":"Department of Urology, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou 510289, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yu","family":"Huang","sequence":"additional","affiliation":[{"name":"Department of Radiology, Shanghai Jiao Tong University School of Medicine Affiliated Shanghai Sixth People\u2019s Hospital, 600 Yi Shan Road, Shanghai 200233, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5975-2026","authenticated-orcid":false,"given":"Kai","family":"Ni","sequence":"additional","affiliation":[{"name":"Department of Urology, Shanghai Jiao Tong University School of Medicine Affiliated Shanghai Sixth People\u2019s Hospital, 600 Yi Shan Road, Shanghai 200233, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xiaoyong","family":"Hu","sequence":"additional","affiliation":[{"name":"Department of Urology, Shanghai Jiao Tong University School of Medicine Affiliated Shanghai Sixth People\u2019s Hospital, 600 Yi Shan Road, Shanghai 200233, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2026,2,15]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Haier, J., and Schaefers, J. 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