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沉默神经纤毛蛋白-1 对结肠癌 SW480 细胞转化生长因子-β1 介导的上皮-间充质转化的影响及其对结肠癌细胞增殖和迁移的影响。

The effect of neuropilin-1 silencing on the transforming growth factor-β1-mediated epithelial-mesenchymal transition of colon cancer SW480 cells and its effect on the proliferation and migration of colon cancer cells.

机构信息

Department of Surgical Oncology, North China University of Science and Technology Affiliated Hospital, Tangshan, China.

Department of General Surgery, North China University of Science and Technology Affiliated Hospital, Tangshan, China.

出版信息

J Physiol Pharmacol. 2022 Apr;73(2). doi: 10.26402/jpp.2022.2.07. Epub 2022 Sep 29.

Abstract

This study aimed to investigate the effect of neuropilin-1 (NRP-1) silencing on epithelial-mesenchymal transformation (EMT) mediated by transforming growth factor-β1 (TGF-β1) and on the proliferation and migration of colon cancer SW480 cells. After transfection of small interfering ribonucleic acid (siRNA)-NRP-1 into colon cancer SW480 cells, the messenger RNA (mRNA) and protein expression levels of NRP-1 were detected using quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot. Four EMT models were induced using 0, 2, 5, and 10 ng/mL TGF-β1, respectively. Cell proliferation was detected using Cell Counting Kit-8, and the protein levels of EMT markers E-cadherin and vimentin were detected using Western blot. EMT was induced in the transfected SW480 cells using TGF-β1, after which four groups were created: a negative control group (siRNA-Ncontrol), a transfection group (siRNA-NRP-1), an induction group (TGF-β1), and a transfection + induction group (siRNA-NRP-1+TGF-β1). Western blot was then used to detect the expression of E-cadherin and vimentin, and cell proliferation and migration were detected using cell counting kit-8 (CCK-8) and scratch assay. After transfection with siRNA-NRP-1, the mRNA and protein expression levels of SW480 cells were significantly decreased (P<0.05). After 48 hours of induction with 10 ng/mL TGF-β1, cell proliferation was obvious, E-cadherin expression decreased, and vimentin expression significantly increased (P<0.05), indicating that EMT had been successfully induced compared with the induction group, the transfection + induction group had significantly increased E-cadherin expression after corresponding treatments (including transfection and induction alone) (P<0.05), and the proliferation and migration of colon cancer cells decreased (P<0.05). In conclusion: silencing, NRP-1 in colon cancer SW480 cells can partially reverse TGF-β1-mediated EMT, reduce the proliferation activity of colon cancer cells, and slow their migration ability. Therefore, NRP-1 may become a new target for the treatment of colon cancer.

摘要

本研究旨在探讨神经纤毛蛋白-1(NRP-1)沉默对转化生长因子-β1(TGF-β1)介导的上皮-间充质转化(EMT)以及结肠癌细胞 SW480 增殖和迁移的影响。用小干扰核糖核酸(siRNA)-NRP-1 转染结肠癌细胞 SW480 后,采用实时定量聚合酶链反应(qRT-PCR)和 Western blot 检测 NRP-1 的信使 RNA(mRNA)和蛋白表达水平。分别用 0、2、5 和 10 ng/mL TGF-β1 诱导 4 种 EMT 模型。采用细胞计数试剂盒-8(Cell Counting Kit-8,CCK-8)检测细胞增殖,Western blot 检测 EMT 标志物 E-钙黏蛋白和波形蛋白的蛋白水平。用 TGF-β1 诱导转染的 SW480 细胞发生 EMT,然后创建 4 个组:阴性对照组(siRNA-Ncontrol)、转染组(siRNA-NRP-1)、诱导组(TGF-β1)和转染+诱导组(siRNA-NRP-1+TGF-β1)。然后用 Western blot 检测 E-钙黏蛋白和波形蛋白的表达,用 CCK-8 和划痕实验检测细胞增殖和迁移。转染 siRNA-NRP-1 后,SW480 细胞的 mRNA 和蛋白表达水平显著降低(P<0.05)。用 10 ng/mL TGF-β1 诱导 48 小时后,细胞增殖明显,E-钙黏蛋白表达减少,波形蛋白表达显著增加(P<0.05),表明与诱导组相比 EMT 成功诱导,转染+诱导组在单独转染和诱导后 E-钙黏蛋白表达显著增加(P<0.05),结肠癌细胞的增殖和迁移减少(P<0.05)。结论:沉默结肠癌细胞 SW480 中的 NRP-1 可部分逆转 TGF-β1 介导的 EMT,降低结肠癌细胞的增殖活性,减缓其迁移能力。因此,NRP-1 可能成为结肠癌治疗的新靶点。

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