PDF(Pubmed)
Abstract：
Drug resistance is a key factor underlying the failure of tumor chemotherapy. It enhances the stem‑like cell properties of cancer cells, tumor metastasis and relapse. Luteolin is a natural flavonoid with strong anti‑tumor effects. However, the mechanism(s) by which luteolin protects against paclitaxel (PTX)‑resistant cancer cell remains to be elucidated. The inhibitory effect of luteolin on the proliferation of EC1/PTX and EC1 cells was detected by cell counting kit‑8 assay. Colony formation and flow cytometry assays were used to assess clonogenic capacity, cell cycle and apoptosis. Wound healing and Transwell invasion tests were used to investigate the effects of luteolin on the migration and invasion of EC1/PTX cells. Western blotting was used to detect the protein levels of EMT‑related proteins and stem cell markers after sphere formation. Parental cells and drug‑resistant cells were screened by high‑throughput sequencing to detect the differential expression of RNA and differential genes. ELISA and western blotting were used to verify the screened PI3K/Akt signaling pathway, key proteins of which were explored by molecular docking. Hematoxylin and eosin staining and TUNEL staining were used to observe tumor xenografts on morphology and apoptosis in nude mice. The present study found that luteolin inhibited tumor resistance (inhibited proliferation, induced cell cycle arrest and apoptosis and hindered migration invasion, EMT and stem cell spherification) in vitro in PTX‑resistant esophageal squamous cell carcinoma (ESCC) cells. In addition, luteolin enhanced drug sensitivity and promoted the apoptosis of drug‑resistant ESCC cells in combination with PTX. Mechanistically, luteolin may inhibit the PI3K/AKT signaling pathway by binding to the active sites of focal adhesion kinase (FAK), Src and AKT. Notably, luteolin lowered the tumorigenic potential of PTX‑resistant ESCC cells but did not show significant toxicity in vivo. Luteolin enhanced drug chemosensitivity by downregulating the FAK/PI3K/AKT pathway in PTX‑resistant ESCC and could be a promising agent for the treatment of PTX‑resistant ESCC cancers.
摘要：
耐药性是导致肿瘤化疗失败的关键因素。它增强了癌细胞的干细胞特性，肿瘤转移和复发。木犀草素是一种天然黄酮类化合物，具有很强的抗肿瘤作用。然而,木犀草素对紫杉醇(PTX)耐药癌细胞的保护机制尚待阐明.通过细胞计数试剂盒-8法检测木犀草素对EC1/PTX和EC1细胞增殖的抑制作用。集落形成和流式细胞术测定用于评估克隆形成能力，细胞周期和细胞凋亡。采用创面愈合和Transwell侵袭试验研究木犀草素对EC1/PTX细胞迁移和侵袭的影响。Western印迹用于检测球体形成后EMT相关蛋白和干细胞标志物的蛋白水平。通过高通量测序筛选亲本细胞和耐药细胞，以检测RNA和差异基因的差异表达。采用ELISA和免疫印迹对筛选出的PI3K/Akt信号通路进行验证,通过分子对接探索了其中的关键蛋白质。采用苏木精、伊红染色和TUNEL染色观察裸鼠移植瘤的形态和凋亡。本研究发现木犀草素抑制肿瘤耐药(抑制增殖,诱导细胞周期阻滞和凋亡，并阻碍迁移侵袭，EMT和干细胞球化）在PTX抗性食管鳞状细胞癌（ESCC）细胞中的体外研究。此外,木犀草素与PTX联合可增强耐药ESCC细胞的药物敏感性并促进其凋亡。机械上,木犀草素可能通过与粘着斑激酶(FAK)的活性位点结合而抑制PI3K/AKT信号通路,Src和AKT。值得注意的是,木犀草素降低了PTX耐药ESCC细胞的致瘤潜能，但在体内未显示出明显的毒性。木犀草素通过下调PTX耐药ESCC中的FAK/PI3K/AKT途径增强药物化学敏感性，可能是治疗PTX耐药ESCC癌症的有前途的药物。
