Department of Pharmacology and Physiology, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok, Thailand.
Cell-Based Drug and Health Product Development Research Unit, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok, Thailand.
Anticancer Res. 2021 Dec;41(12):6147-6154. doi: 10.21873/anticanres.15434.
BACKGROUND/AIM: Cancer metastasis is the main cause of mortality in cancer patients. As lung cancer patients are mostly detected at metastatic stages, strategies that inhibit cancer metastasis may offer effective therapies. Activation of FAK and Akt/mTOR pathways promotes the highly metastatic phenotypes of epithelial to mesenchymal transition (EMT). We unraveled EMT inhibitory action of pongamol and the mechanism controlling cell dissemination in lung cancer cells.
Cytotoxic and antiproliferative effects of pongamol were determined by the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay. Apoptosis and necrosis induction in response to pongamol treatment was observed and visualized by nuclei staining assay. Wound healing migration, invasion, and anchorage-dependent growth assay were conducted to evaluate metastatic behaviors. EMT protein expression and FAK pathway were detected by western blot analysis.
Pongamol at 0-100 μM exhibited significant inhibition on migration, and invasion of cancer cells. Regarding anoikis resistance potential, the compound significantly inhibited survival and growth of cancer cells in an anchorage-independent manner, as indicated by the depletion of growing colonies in pongamol-pretreated cells. Protein level analysis further showed that pongamol exerted its anti-metastasis effect by inhibiting EMT, as indicated by a decrease of several mesenchymal proteins (N-cadherin, vimentin, Snail, and Slug). Regarding the up-stream mechanisms, we found that pongamol inhibited activation of FAK and Akt/mTOR signaling pathways.
Pongamol exhibits potent anti-metastatic activity through suppressing key potentiating factors of cancer metastasis EMT and FAK.
背景/目的：癌症转移是癌症患者死亡的主要原因。由于肺癌患者大多在转移阶段被发现，因此抑制癌症转移的策略可能提供有效的治疗方法。FAK 和 Akt/mTOR 通路的激活促进了上皮间质转化（EMT）的高度转移性表型。我们揭示了蓬甘醇抑制 EMT 的作用及其控制肺癌细胞细胞扩散的机制。
通过 3-[4,5-二甲基噻唑-2-基]-2,5-二苯基四氮唑溴盐（MTT）测定法测定蓬甘醇的细胞毒性和抗增殖作用。通过核染色测定观察和可视化蓬甘醇处理后细胞凋亡和坏死的诱导。进行划痕愈合迁移、侵袭和锚定依赖性生长测定以评估转移行为。通过 Western blot 分析检测 EMT 蛋白表达和 FAK 通路。
蓬甘醇在 0-100 μM 时对癌细胞的迁移和侵袭表现出显著的抑制作用。关于抗凋亡抵抗潜力，该化合物通过在无锚定生长测定中抑制癌细胞的存活和生长来显著抑制，这表明在蓬甘醇预处理的细胞中生长菌落的减少。蛋白质水平分析进一步表明，蓬甘醇通过抑制 EMT 发挥其抗转移作用，这表现为几种间质蛋白（N-钙粘蛋白、波形蛋白、Snail 和 Slug）的减少。关于上游机制，我们发现蓬甘醇抑制了 FAK 和 Akt/mTOR 信号通路的激活。
蓬甘醇通过抑制癌症转移 EMT 和 FAK 的关键增强因子表现出强大的抗转移活性。
