Department of Surgical Oncology, Ningbo No.2 Hospital, Ningbo 315010, P.R. China.
School of Nursing, Wenzhou Medical University, Wenzhou 325000, P.R. China.
Mol Cells. 2018 Sep 30;41(9):853-867. doi: 10.14348/molcells.2018.0103. Epub 2018 Aug 31.
As the most common type of endocrine malignancy, papillary thyroid cancer (PTC) accounts for 85-90% of all thyroid cancers. In this study, we presented the hypothesis that SDC4 gene silencing could effectively attenuate epithelial mesenchymal transition (EMT), and promote cell apoptosis the Wnt/β-catenin signaling pathway in human PTC cells. Bioinformatics methods were employed to screen the determined differential expression levels of SDC4 in PTC and adjacent normal samples. PTC tissues and adjacent normal tissues were prepared and their respective levels of SDC4 protein positive expression, in addition to the mRNA and protein levels of SDC4, Wnt/β-catenin signaling pathway, EMT and apoptosis related genes were all detected accordingly. Flow cytometry was applied in order to detect cell cycle entry and apoptosis. Finally, analyses of PTC migration and invasion abilities were assessed by using a Transwell assay and scratch test. In PTC tissues, activated Wnt/β-catenin signaling pathway, increased EMT and repressed cell apoptosis were determined. Moreover, the PTC K1 and TPC-1 cell lines exhibiting the highest SDC4 expression were selected for further experiments. experiments revealed that SDC4 gene silencing could suppress cell migration, invasion and EMT, while acting to promote the apoptosis of PTC cells by inhibiting the activation of the Wnt/β-catenin signaling pathway. Besides, si-β-catenin was observed to inhibit the promotion of PTC cell migration and invasion caused by SDC4 overexpression. Our study revealed that SDC4 gene silencing represses EMT, and enhances cell apoptosis by suppressing the activation of the Wnt/β-catenin signaling pathway in human PTC.
作为最常见的内分泌恶性肿瘤,甲状腺乳头状癌(PTC)占所有甲状腺癌的 85-90%。在这项研究中,我们提出了这样的假设,即 SDC4 基因沉默可以有效减弱上皮间质转化(EMT),并促进人 PTC 细胞中 Wnt/β-catenin 信号通路的细胞凋亡。采用生物信息学方法筛选出 PTC 与相邻正常样本中确定的 SDC4 差异表达水平。制备 PTC 组织和相邻正常组织,并相应检测 SDC4 蛋白阳性表达水平,以及 SDC4、Wnt/β-catenin 信号通路、EMT 和细胞凋亡相关基因的 mRNA 和蛋白水平。流式细胞术用于检测细胞周期进入和细胞凋亡。最后,通过 Transwell 测定和划痕试验评估 PTC 迁移和侵袭能力。在 PTC 组织中,确定了激活的 Wnt/β-catenin 信号通路、增加的 EMT 和抑制的细胞凋亡。此外,选择 SDC4 表达最高的 PTC K1 和 TPC-1 细胞系进行进一步实验。实验表明,SDC4 基因沉默可通过抑制 Wnt/β-catenin 信号通路的激活来抑制 PTC 细胞迁移、侵袭和 EMT,同时促进 PTC 细胞凋亡。此外,观察到 si-β-catenin 抑制 SDC4 过表达引起的 PTC 细胞迁移和侵袭的促进作用。我们的研究表明,SDC4 基因沉默通过抑制 Wnt/β-catenin 信号通路的激活来抑制 EMT,并增强人 PTC 中的细胞凋亡。