Oncogene 5'-3' exoribonuclease 2 enhances epidermal growth factor receptor signaling pathway to promote epithelial-mesenchymal transition and metastasis in non-small-cell lung cancer.

IF 2.5 4区医学 Q2 PATHOLOGY

Cytojournal Pub Date : 2024-11-19 eCollection Date: 2024-01-01 DOI:10.25259/Cytojournal_49_2024

Yonghui Cheng, Mengge Wen, Xiaochun Wang, Hao Zhu

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引用次数: 0

Abstract

Objective: Epithelial-mesenchymal transition (EMT) and metastasis are the primary causes of mortality in non-small-cell lung cancer (NSCLC). 5'-3' exoribonuclease 2 (XRN2) plays an important role in the process of tumor EMT. Thus, this investigation mainly aimed to clarify the precise molecular pathways through which XRN2 contributes to EMT and metastasis in NSCLC.

Material and methods: Western blot and quantitative real-time polymerase chain reaction were first used to assess XRN2 levels in NSCLC cells. Subsequently, short hairpin RNA-XRN2 (Sh-XRN2) and XRN2 overexpression (Ov-XRN2) plasmids were transfected to NSCLC cells. The effects of Sh-XRN2 and Ov-XRN2 on NSCLC cell migration and invasion were evaluated by Transwell assay. Western blot experiments were conducted to assess the effects of Sh-XRN2 and Ov-XRN2 on proteins related to EMT and the epidermal growth factor receptor (EGFR) signaling pathway in H460 cells. Then, Sh-XRN2 and EGFR overexpression (Ov-EGFR) plasmids were transfected to NSCLC cells. Changes in NSCLC cell migration and invasion were measured using a Transwell assay with Sh-XRN2 and Sh-XRN2+Ov-EGFR. Changes in the expression of proteins related to EMT in NSCLC cells were detected by Western blot assays with Sh-XRN2 and Sh-XRN2+Ov-EGFR. Furthermore, a subcutaneous tumor model for NSCLC was established. Immunohistochemical analysis was performed to assess the levels of Cluster of Differentiation 31 (CD31) in lung metastatic lesions. H460 cells transfected with Sh-XRN2, Ov-XRN2 or Sh-XRN2+Ov-EGFR were co-cultured with human umbilical vein endothelial cells (HUVECs) to assess the tube formation ability of the cells.

Results: Compared with those observed in human bronchial epithelial cells (BEAS-2B cells), XRN2 expression levels were significantly upregulated in NSCLC cell lines (H460 cells) (P < 0.001). XRN2 overexpression considerably promoted the NSCLC cell migration and invasion, EMT process, and tube formation ability of HUVECs (P < 0.001). On the contrary, XRN2 knockdown led to a reduction in these processes. In addition, XRN2 overexpression increased the expression levels of CD31 in lung metastatic lesions and activated the phosphorylation of EGFR signaling pathway (P < 0.001). Furthermore, Sh-XRN2+Ov-EGFR significantly promoted migration, invasion, and EMT processes in H460 cells (P < 0.001). In the meantime, compared with the co-H460+Sh-XRN2+Ov-NC group, co-H460+Sh-XRN2+Ov-EGFR significantly enhanced the tube formation ability of HUVECs (P < 0.001).

Conclusion: XRN2 promoted EMT and metastasis in NSCLC through improving the phosphorylation of the EGFR signaling pathway in NSCLC cells.

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癌基因5′-3′外核糖核酸酶2增强表皮生长因子受体信号通路促进非小细胞肺癌上皮-间质转化和转移

目的：上皮-间质转化（EMT）和转移是导致非小细胞肺癌（NSCLC）死亡的主要原因。5‘-3’外核糖核酸酶2 （XRN2）在肿瘤EMT过程中起重要作用。因此，本研究主要旨在明确XRN2在NSCLC中参与EMT和转移的确切分子途径。材料和方法：首次采用Western blot和实时定量聚合酶链反应（pcr）技术检测NSCLC细胞中的XRN2水平。随后，将短发夹RNA-XRN2 （Sh-XRN2）和XRN2过表达（Ov-XRN2）质粒转染至NSCLC细胞。Transwell法观察Sh-XRN2和Ov-XRN2对NSCLC细胞迁移和侵袭的影响。Western blot实验评估Sh-XRN2和Ov-XRN2对H460细胞EMT相关蛋白和表皮生长因子受体（EGFR）信号通路的影响。然后将Sh-XRN2和EGFR过表达（Ov-EGFR）质粒转染至NSCLC细胞。采用Sh-XRN2和Sh-XRN2+Ov-EGFR的Transwell实验测量NSCLC细胞迁移和侵袭的变化。用Sh-XRN2和Sh-XRN2+Ov-EGFR检测NSCLC细胞中EMT相关蛋白的表达变化。建立非小细胞肺癌皮下肿瘤模型。采用免疫组化方法检测肺转移灶中CD31的表达水平。将转染Sh-XRN2、Ov-XRN2或Sh-XRN2+Ov-EGFR的H460细胞与人脐静脉内皮细胞（HUVECs）共培养，评估细胞的成管能力。结果：与人支气管上皮细胞（BEAS-2B细胞）相比，XRN2在非小细胞肺癌细胞系（H460细胞）中的表达水平显著上调（P < 0.001）。XRN2过表达显著促进了非小细胞肺癌细胞的迁移侵袭、EMT过程和HUVECs的成管能力（P < 0.001）。相反，XRN2敲低导致这些过程的减少。此外，XRN2过表达增加了肺转移灶中CD31的表达水平，激活了EGFR信号通路的磷酸化（P < 0.001）。此外，Sh-XRN2+Ov-EGFR显著促进H460细胞的迁移、侵袭和EMT过程（P < 0.001）。同时，与co-H460+Sh-XRN2+Ov-NC组相比，co-H460+Sh-XRN2+Ov-EGFR显著增强了HUVECs成管能力（P < 0.001）。结论：XRN2通过改善NSCLC细胞中EGFR信号通路的磷酸化，促进了NSCLC的EMT和转移。

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来源期刊

Cytojournal PATHOLOGY-

CiteScore

2.20

自引率

42.10%

发文量

审稿时长

>12 weeks

期刊介绍： The CytoJournal is an open-access peer-reviewed journal committed to publishing high-quality articles in the field of Diagnostic Cytopathology including Molecular aspects. The journal is owned by the Cytopathology Foundation and published by the Scientific Scholar.