摘要
目的 从细胞水平探讨芪玉三龙方是否通过 Wnt5a/β-catenin信号通路抑制上皮—间充质细胞转化,抑制肺癌转移.方法 将重组慢病毒转染小鼠肺癌细胞建立稳定过表达 Wnt5a的细胞系,同时构建 Wnt5a干扰和空载细胞株.qRT-PCR法检测 Wnt5a、β-catenin mRNA的表达水平.CCK8、EdU实验检测肺癌细胞的增殖活性,Transwell实验检测肺癌细胞的迁移和侵袭能力.Western blot法检测 Wnt5a、β-catenin、c-Myc、Cyclin D1、Vimen-tin、N-cadherin、E-cadherin、Snail蛋白表达水平.结果 与对照组比较,芪玉三龙方组 Wnt5a、β-catenin、c-Myc、Cy-clin D1 蛋白表达水平下降(P＜0.05),细胞增殖活性、侵袭和迁移能力显著降低(P＜0.05).Wnt5a过表达组中Vimentin、N-cadherin、Snail表达水平增加(P＜0.05),E-cadherin蛋白表达水平下降(P＜0.05),细胞增殖活性、侵袭和迁移能力显著增强(P＜0.05);Wnt5a干扰组 Vimentin、N-cadherin、Snail表达水平下降(P＜0.05),E-cadherin蛋白表达水平升高(P＜0.05),细胞增殖活性、侵袭和迁移能力显著下降(P＜0.05),明显抑制 EMT的发生.芪玉三龙方处理后,Wnt5a过表达组 Wnt5a、β-catenin、c-Myc、Cyclin D1、Vimentin、N-cadherin、Snail表达水平下降(P＜0.05),E-cadherin蛋白表达水平升高(P＜0.05),细胞增殖活性、侵袭和迁移能力得到抑制.结论 芪玉三龙方可能通过 Wnt5a/β-catenin信号通路逆转上皮—间充质转化,抑制肺癌细胞增殖和转移.
Abstract
Objective To investigate whether Qiyu Sanlong Prescription can inhibit the metastasis of lung cancer by inhibiting epithelial-mesenchymal transition via the Wnt5a/β-catenin signaling pathway at the cellular level.Methods Mouse lung cancer cells were transfected with recombinant lentivirus to establish a cell line with the stable expression of Wnt5a,and Wnt5a inter-ference and empty vector cell lines were established.qRT-PCR was used to measure the expression of Wnt5a and β-catenin.CCK8 and EdU assay were used to measure the proliferative activity of cells,and Transwell assay was used to measure the mi-gration and invasion abilities of cells.Western blot was used to measure the protein expression levels of Wnt5a,β-catenin,c-Myc,Cyclin D1,Vimentin,N-cadherin,E-cadherin,and Snail.Results Compared with the control group,the Qiyu Sanlong Pre-scription group had significant reductions in the protein expression levels of Wnt5a,β-catenin,c-Myc,and Cyclin D1(P＜0.05)and significantly lower proliferative activity and invasion and migration abilities of cells(P＜0.05).The Wnt5a overexpression group had significant increases in the expression levels of Vimentin,N-cadherin,and Snail(P＜0.05),a significant reduction in the protein expression level of E-cadherin(P＜0.05),and significant enhancement of the proliferative activity and invasion and migration abilities of cells(P＜0.05);the Wnt5a interference group had significant reductions in the expression levels of Vimentin,N-cadherin,and Snail(P＜0.05);a significant increase in the protein expression level of E-cadherin(P＜0.05),and significant reductions in the proliferative activity and invasion and migration abilities of cells(P＜0.05).After treatment with Qiyu Sanlong pre-scription,the Wnt5a overexpression group had significant reductions in the expression levels of Wnt5a,β-catenin,c-Myc,Cyclin D1,Vi-mentin,N-cadherin,and Snail(P＜0.05),a significant increase in the expression level of E-cadherin(P＜0.05),and significant inhibition of the proliferative activity and invasion and migration abilities of cells(P＜0.05).Conclusion Qiyu Sanlong prescription inhibits the proliferation of tumor cells by reversing epithelial-mesenchymal transition via the Wnt5a/β-catenin signaling pathway.
基金项目
国家自然科学基金(81874431)
安徽省高等学校优秀拔尖人才培育项目(gxbjZD2021057)
安徽省高等学校自然科学研究重点项目(KJ2019A0468)
安徽省高等学校自然科学研究重点项目(KJ2021A0542)
维普
万方数据