Masraksa Wuttipong, Tanasawet Supita, Hutamekalin Pilaiwanwadee, Wongtawatchai Tulaporn, Sukketsiri Wanida
Department of Pharmacology, Faculty of Science, Prince of Songkla University, Songkhla, 90110, Thailand.
Department of Anatomy, Faculty of Science, Prince of Songkla University, Songkhla, 90110, Thailand.
Nutr Res Pract. 2020 Apr;14(2):127-133. doi: 10.4162/nrp.2020.14.2.127. Epub 2019 Nov 22.
BACKGROUND/OBJECTIVES: Non-small cell lung cancer is mostly recognized among other types of lung cancer with a poor prognosis by cause of chemotherapeutic resistance and increased metastasis. Luteolin has been found to decrease cell metastasis. However, its underlying mechanisms remain unresolved. The objective of this study was to examine the effect (and its mechanism) of luteolin on the migration and invasion of human non-small cell lung cancer A549 cells.
MATERIALS/METHODS: Cell viability was investigated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Wound healing and transwell assays were evaluated to assess migration and invasion, respectively. Western blot analysis and immunofluorescence were further performed to investigate the role of luteolin and its mechanisms of action.
Administration with up to 40 µM luteolin showed no cytotoxic activity on lung cancer A549 cells or non-cancer MRC-5 cells. Additionally, luteolin at 20-40 µM significantly suppressed A549 cells' migration, invasion, and the formation of filopodia in a concentration-dependent manner at 24 h. This is similar with western blot analysis, which revealed diminished the phosphorylated focal adhesion kinase (pFAK), phosphorylated non-receptor tyrosine kinase (pSrc), Ras-related C3 botulinum toxin substrate 1 (Rac1), cell division control protein 42 (Cdc42), and Ras homolog gene family member A (RhoA) expression levels.
Overall, our data indicate that luteolin plays a role in controlling lung cancer cells' migration and invasion via Src/FAK and its downstream Rac1, Cdc42, and RhoA pathways. Luteolin might be considered a promising candidate for suppressing invasion and metastasis of lung cancer cells.
背景/目的：非小细胞肺癌在其他类型的肺癌中最为常见，因其化疗耐药性和转移增加，预后较差。已发现木犀草素可减少细胞转移。然而，其潜在机制仍未明确。本研究的目的是探讨木犀草素对人非小细胞肺癌A549细胞迁移和侵袭的影响（及其机制）。
材料/方法：采用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐法检测细胞活力。分别通过伤口愈合试验和Transwell试验评估迁移和侵袭能力。进一步进行蛋白质免疫印迹分析和免疫荧光检测，以研究木犀草素的作用及其作用机制。
给予高达40 μM的木犀草素对肺癌A549细胞或非癌MRC-5细胞均无细胞毒性活性。此外，20 - 40 μM的木犀草素在24小时时以浓度依赖的方式显著抑制A549细胞的迁移、侵袭和丝状伪足的形成。这与蛋白质免疫印迹分析结果相似，该分析显示磷酸化粘着斑激酶（pFAK）、磷酸化非受体酪氨酸激酶（pSrc）、Ras相关C3肉毒杆菌毒素底物1（Rac1）、细胞分裂控制蛋白42（Cdc42）和Ras同源基因家族成员A（RhoA）的表达水平降低。
总体而言，我们的数据表明木犀草素通过Src/FAK及其下游的Rac1、Cdc42和RhoA途径在控制肺癌细胞的迁移和侵袭中发挥作用。木犀草素可能被认为是抑制肺癌细胞侵袭和转移的有前途的候选药物。
