异补骨脂查耳酮对鼻咽癌CNE1细胞增殖和转移的影响及作用机制的研究

Effects and Mechanism of Isobavachalcone on the Proliferation and Metastasis of Nasopharyngeal Carcinoma CNE1 Cells

目的:探讨异补骨脂查耳酮对鼻咽癌CNE1细胞增殖和转移的影响及作用机制。方法:采用CCK-8、克隆平板、划痕和Transwell实验检测不同浓度(10,20,40μmol·L^(-1))异补骨脂查耳酮对鼻咽癌CNE1细胞增殖、克隆形成、迁移和侵袭能力的影响;利用生物信息学技术分析异补骨脂查耳酮的核心作用靶点;Western Blot检测异补骨脂查耳酮对其核心作用靶点表皮生长因子(EGFR)及其下游通路蛋白磷脂酰肌醇-3-羟激酶(PI3K)、磷酸化磷脂酰肌醇-3-羟激酶(p-PI3K)、蛋白激酶B(AKT)、磷酸化蛋白激酶B(p-AKT)和雷帕霉素靶蛋白(mTOR)蛋白的影响。CNE-1细胞分别予以DMSO、40μmol·L^(-1)异补骨脂查耳酮、40μmol·L^(-1)异补骨脂查耳酮联合100μmol·L^(-1)EGFR激动剂NSC 228155以及40μmol·L^(-1)异补骨脂查耳酮联合10μmol·L^(-1)AKT激动剂SC79处理,应用CCK-8、克隆平板、划痕及Transwell实验检测各组细胞的增殖、克隆形成、迁移和侵袭能力的改变。结果:异补骨脂查耳酮显著抑制鼻咽癌CNE1细胞的增殖、克隆形成、迁移和侵袭能力(P<0.05或P<0.01)。通过生物信息学分析,异补骨脂查耳酮有104个作用靶点,作用靶点主要富集在肿瘤发生通路上;其中,作用靶点EGFR与其他靶点蛋白联系最广泛,且在多个通路上均有富集,被认为是异补骨脂查耳酮的核心作用靶点。Western Blot结果显示,异补骨脂查耳酮显著抑制鼻咽癌CNE1细胞中EGFR、p-PI3K、p-AKT和mTOR蛋白的表达(P<0.05或P<0.01)。此外,EGFR激动剂NSC228155和AKT激动剂SC79能够显著减轻异补骨脂查耳酮介导的CNE-1细胞增殖、克隆形成、迁移和侵袭能力的抑制(P<0.01)。结论:异补骨脂查耳酮可能是通过抑制EGFR/PI3K/AKT/mTOR信号通路减少鼻咽癌CNE1细胞增殖和转移能力。

Objective:To investigate the effects and mechanism of isobavachalcone on the proliferation and metastasis of nasopharyngeal carcinoma CNE1 cells.Methods:CCK-8,colony formation assay,wound healing and Transwell assay were used to detect the effects of 0,10,20 and 40μmol·L^(-1)isobavachalcone on the proliferation,colony formation,migration and invasion of nasopharyngeal carcinoma CNE1 cells;the bioinformatics method was performed to analyze the core target of isobavachalcone;Western blot was used to detect the expression of core target EGFR of isobavachalcone in nasopharyngeal carcinoma CNE1 cells,as well as the downstream proteins of EGFR such as PI3 K,p-PI3 K,AKT,p-AKT and mTOR.CNE-1 was treated with DMSO,40μmol·L^(-1)isobavachalcone,40μmol·L^(-1)isobavachalcone plus 100μmol·L^(-1)EGFR activator NSC228155 and 40μmol·L^(-1)isobavachalcone plus 10μmol·L^(-1)AKT activator SC79,and CCK-8,colony formation,wound healing and transwell assay were used to detect the proliferation,colony formation,migration and invasion in each group.Results:Isobavachalcone significantly decreased the proliferation,colony formation,migration and invasion of nasopharyngeal carcinoma CNE1 cells(P<0.05 or P<0.01).Bioinformatics results demonstrated that there were 104 protein targets for isobavachalcone,and the targets were most enriched in pathways in cancer.Among them,EGFR interacted with the protein targets comprehensively and enriched in a series of pathways,therefore,EGFR was set as the core target of isobavachalcone.Western blot results indicated that isobavachalcone significantly decreased the expressions of EGFR,p-PI3 K,p-AKT and mTOR(P<0.05 or P<0.01).Moreover,EGFR activator NSC228155 and AKT activator SC79 could significantly decrease the inhibitory effects of isobavachalcone on the proliferation,colony formation,migration and invasion of CNE-1 cell(P<0.01).Conclusion:Isobavachalcone may decrease the proliferation and metastasis of nasopharyngeal carcinoma CNE1 cells by EGFR/PI3 K/AKT/mTOR pathway.