背景:卵巢癌(OVCA)是最致命的妇科恶性肿瘤之一。和厚朴酚(HNK)对包括OVCA在内的恶性肿瘤的抗肿瘤活性已被大量研究证实。因此,这项工作旨在阐明HNK介导的YAP/TAZ通路调节对OVCA细胞生物学功能的影响.
方法:用不同浓度(0、25、50、75和100μM)的HNK处理OVCA细胞,伴随YAP激动剂(XMU)的给药。使用CCK-8测定进行细胞活力的评估,同时通过集落形成测定法对细胞增殖进行定量。使用流式细胞术确定细胞凋亡,和凋亡相关蛋白(caspase-3,Bcl-2,Bax)的表达,EMT相关蛋白(E-cadherin,N-钙黏着蛋白),迁移相关蛋白(MMP-2,MMP-9),和YAP/TAZ途径相关蛋白通过蛋白质印迹进行评估。进行Transwell实验以评估细胞迁移和侵入性倾向。建立移植瘤模型观察肿瘤生长(体积和重量),通过TUNEL染色评估细胞凋亡,通过IHC评估Ki67表达。
结果:HNK对OVCA细胞的活力和增殖能力有抑制作用,引起的凋亡反应,减少了细胞的迁移和侵袭倾向,并下调YAP/TAZ途径。用YAP激动剂(XMU-MP-1)刺激部分减弱HNK对OVCA细胞生物学的影响。体内实验证实HNK抑制OVCA肿瘤生长。
结论:这项研究的结果最终确定了HNK协调了YAP/TAZ途径的调节,从而控制OVCA细胞的恶性表型表现。确定的HNK在抑制细胞增殖和肿瘤进展中的功能提供了其在OVCA细胞内的抗增殖活性的新证据。
BACKGROUND: Ovarian cancer (OVCA) stands as one of the most fatal gynecological malignancies. Honokiol (HNK) has been substantiated by numerous studies for its anti-tumor activity against malignancies including OVCA. Consequently, this work was designed to elucidate the impact of HNK-mediated modulation of the YAP/TAZ pathway on the biological functions of OVCA cells.
METHODS: OVCA cells were subjected to treatment with varying concentrations (0, 25, 50, 75, and 100 μM) of HNK, concomitant with the administration of YAP agonist (XMU). Assessment of cellular viability was executed employing the CCK-8 assay, while quantification of cellular proliferation transpired via colony formation assays. Apoptosis was ascertained using flow cytometry, and expression of apoptosis-related proteins (caspase-3, Bcl-2, Bax), EMT-related proteins (E-cadherin, N-cadherin), migration-associated proteins (MMP-2, MMP-9), and YAP/TAZ pathway-related proteins was evaluated by western blot. Transwell experiments were conducted to assess cellular migratory and invasive propensities. Xenograft tumor models were built to observe tumor growth (volume and weight), apoptosis was assessed by TUNEL staining, and Ki67 expression was evaluated through IHC.
RESULTS: HNK exerted inhibitory effects on the viability and proliferative capacity of OVCA cells, elicited apoptotic responses, curtailed the migratory and invasive tendencies of cells, and downregulated the YAP/TAZ pathway. Stimulation with YAP agonist (XMU-MP-1) partially attenuated the impacts of HNK on OVCA cell biology. Experiments in vivo confirmed that HNK inhibited OVCA tumor growth.
CONCLUSIONS: The outcomes of this investigation conclusively established that HNK orchestrated the modulation of the YAP/TAZ pathway, thereby exerting control over the malignant phenotypic manifestations of OVCA cells. The ascertained function of HNK in restraining cellular proliferation and tumor progression provided novel evidence of its anti-proliferative activity within OVCA cells.