摘要
目的:运用小干扰RNA技术下调表达β-catenin来评价Wnt/β-catenin信号通路在TGF-β1诱导肝癌细胞发生生物学行为改变中的作用。方法:首先通过蛋白印迹实验验证siRNA-β-catenin的干扰效果;肝癌细胞转染对照质粒和siRNA-β-catenin后分别加入细胞因子TGF-β1共培养,再采用MTT、细胞划痕实验、Transwell实验检测肝癌细胞增殖、迁移、侵袭等生物学行为的改变;蛋白印迹实验检测转染siRNA-β-catenin后加入细胞因子TGF-β1诱导的肝癌细胞中EMT相关标志物蛋白表达变化。结果:siRNA-β-catenin能够显著下调肝癌细胞中β-catenin蛋白的表达;MTT结果显示:siRNA-β-catenin转染肝癌细胞后能够显著抑制TGF-β1诱导的促进肝癌细胞增殖的作用;细胞划痕实验和Transwell实验也证实转染了siRNA-β-catenin的肝癌细胞能够部分抵抗TGF-β1诱导的促进细胞迁移和侵袭的作用;蛋白印迹实验结果显示siRNA-β-catenin转染肝癌细胞后,能够抑制TGF-β1诱导的EMT样改变。这些结果提示我们β-catenin可能是TGF-β1诱导肝癌细胞发生EMT样改变影响其增殖、迁移、侵袭等生物学行为中的关键蛋白。结论:下调肝癌细胞中β-catenin蛋白表达能够部分抑制TGF-β1诱导的EMT样改变,提示β-catenin可能是TGF-β1诱导肝癌细胞发生EMT样改变中的关键蛋白分子。
Objective:To clarify whether siRNA-β-catenin could inhibit the proliferation,invasion and metastasis SMMC-7721 induced by TGF-β1.Methods:First Western blot experiments were used to detect the interference effect of siRNA-β-catenin;hepatoma cells were transfected with control plasmid and siRNA-β-catenin when added cytokine TGF-β1 were co-cultured,and then used MTT,wound-healing assay,transwell experiments used to detect cell proliferation,migration,invasion changes;western blot assays were used to detect EMT-related markers after transfection of siRNA-β-catenin meanwhile added cytokine TGF-β1.Results:siRNA-β-catenin significantly down-regulated expression ofβ-catenin protein in liver cancer cell SMMC-7721;MTT results showed that siRNA-β-catenin transfected hepatoma cells can significantly inhibit TGF-β1-induced promotion of liver cancer cell proliferation;cell scratch experiment and Transwell experiments also confirmed transfected with siRNA-β-catenin in hepatoma cells can be partially resistant to TGF-β1-induced promote cell migration and invasion;after Western blot results showed that siRNA-β-catenin transfected hepatoma cells,can inhibition of TGF-β1-induced EMT-like changes,showing the expression of EMT-related marker proteins.These results suggest thatβ-catenin might be a key protein in TGF-β1-induced liver cancer cells EMT-like changes.Conclusions:Down-regulatedβ-catenin could inhibit the proliferation,invasion and metastasis of liver cancer cells and the possible mechanism was Wnt/β-catenin signal pathway.
作者
吴志超
王海全
乔晟先
WU Zhi-chao;WANG Hai-quan;QIAO Sheng-xian(Department of Hepatobiliary Surgery,the Second Clinical Medicine School of Qingdao University Qingdao Central Hostipal,Qingdao 266042,China)
出处
《中国现代普通外科进展》
CAS
2021年第10期768-772,共5页
Chinese Journal of Current Advances in General Surgery
基金
青岛市卫计委2017年医药科研计划入选科研课题(2017-WJZD056)。
