14-3-3σ 蛋白促进肝细胞生长因子介导的人胃癌细胞增殖和侵袭基质金属蛋白酶-1 的调节。

14-3-3 Sigma Protein Contributes to Hepatocyte Growth Factor-mediated Cell Proliferation and Invasion Matrix Metalloproteinase-1 Regulation in Human Gastric Cancer.

机构信息

Department of Hematology-Oncology, College of Medicine, Yeungnam University, Daegu, Republic of Korea.

Department of Biochemistry and Molecular Biology, College of Medicine, Yeungnam University, Daegu, Republic of Korea

出版信息

Anticancer Res. 2022 Jan;42(1):519-530. doi: 10.21873/anticanres.15510.

DOI:10.21873/anticanres.15510

PMID:34969762

Abstract

BACKGROUND/AIM: The 14-3-3 protein family has a variety of functions in cellular responses in different organisms, including cell-cycle regulation, apoptosis, and malignant transformation. 14-3-3 Sigma protein (14-3-3σ) induces G2 arrest, which enables repair of damaged DNA. The purpose of this study was to identify the role of 14-3-3σ up-regulation by hepatocyte growth factor (HGF) in cancer cell proliferation and invasion in gastric cancer.

MATERIALS AND METHODS

In this study, cell culture, western blotting, real-time polymerase chain reaction, zymography, 14-3-3σ knock-down using short hairpin RNA (shRNA), electrophoresis mobility-shift assay, chromatin immunoprecipitation assay and standard two-chamber invasion assay were applied.

RESULTS

Firstly, we confirmed that the expression of 14-3-3σ in gastric cancer cells was up-regulated by HGF. To identify how HGF-induced 14-3-3σ expression affects matrix metalloproteinase-1 (MMP1) expression, the cells were treated with the mitogen-activated protein kinase kinase inhibitor PD098059 and analyzed using western blotting. The HGF-mediated expression of MMP1 protein decreased in the presence of PD098059. The role of 14-3-3σ in MMP1 expression was determined through 14-3-3σ knockdown using shRNA. 14-3-3σ-shRNA cells showed reduced levels of MMP1, phosphorylated extracellular signal-regulated kinase, and pp38. HGF-mediated cell proliferation and in vitro invasion were reduced in 14-3-3σ knockdown cells. Serum 14-3-3σ levels were also significantly reduced following gastrectomy in patients with stage II or stage III gastric cancer (p<0.05).

CONCLUSION

These results suggest that 14-3-3σ plays an important role in cell proliferation and metastasis in gastric cancer, and 14-3-3σ may be a novel target for detection and prevention of progression of gastric cancer. In addition, the serum 14-3-3σ level is associated with treatment status in patients with locally advanced gastric cancer.

摘要

背景/目的：14-3-3 蛋白家族在不同生物体的细胞反应中具有多种功能，包括细胞周期调控、细胞凋亡和恶性转化。14-3-3σ 蛋白（14-3-3σ）诱导 G2 期阻滞，从而促进受损 DNA 的修复。本研究旨在确定肝细胞生长因子（HGF）上调 14-3-3σ 在胃癌细胞增殖和侵袭中的作用。

材料和方法

在这项研究中，应用细胞培养、western blot、实时聚合酶链反应、酶谱分析、短发夹 RNA（shRNA）敲低 14-3-3σ、电泳迁移率变动分析、染色质免疫沉淀分析和标准双层侵袭分析。

结果

首先，我们证实了 HGF 上调胃癌细胞中 14-3-3σ 的表达。为了确定 HGF 诱导的 14-3-3σ 表达如何影响基质金属蛋白酶-1（MMP1）的表达，用丝裂原活化蛋白激酶激酶抑制剂 PD098059 处理细胞，并进行 western blot 分析。在 PD098059 存在的情况下，HGF 介导的 MMP1 蛋白表达减少。通过 shRNA 敲低 14-3-3σ 确定 14-3-3σ 在 MMP1 表达中的作用。14-3-3σ-shRNA 细胞显示 MMP1、磷酸化细胞外信号调节激酶和 pp38 的水平降低。14-3-3σ 敲低细胞中 HGF 介导的细胞增殖和体外侵袭减少。在 II 期或 III 期胃癌患者行胃切除术后，血清 14-3-3σ 水平也显著降低（p<0.05）。