EGFL7 promotes hepatocellular carcinoma cell proliferation and inhibits cell apoptosis through increasing CKS2 expression by activating Wnt/β-catenin signaling
EGFL7 promotes hepatocellular carcinoma cell proliferation and inhibits cell apoptosis through increasing CKS2 expression by activating Wnt/β-catenin signaling
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EGFL7 通过激活 Wnt/β-catenin 信号传导增加 CKS2 表达，促进肝细胞癌细胞增殖并抑制细胞凋亡
DOI：
10.1002/jcb.27375
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发表时间：
2018-12-01
影响因子：
4
通讯作者：
Ni, Cai-Fang
中科院分区：
文献类型：
--
作者：
Li, Zhi;Xue, Tong-Qing;Ni, Cai-Fang
表皮生长因子样结构域多聚体7(EGFL7)是一种重要的运动刺激因子，运动相关因子显著促进肿瘤细胞的转移，并在包括肝细胞癌在内的多种肿瘤中过表达，与肿瘤的发生有关。然而，EGFL7调控肝癌细胞增殖和凋亡的分子机制及其与细胞周期蛋白依赖性蛋白激酶调节亚基2(CKS2)之间的关系尚未完全阐明。本研究采用实时定量聚合酶链式反应和免疫组织化学方法检测了肝细胞癌组织中EGFL7和CKS2的表达。将pLKO.1-EGFL7-shRNA、pLVX-Puro-EGFL7重组载体和CKS2小干扰RNA分别转染肝癌细胞后，分别用细胞计数试剂盒8和流式细胞仪检测细胞增殖和凋亡情况，并用Western印迹法检测β-连环蛋白、CKS2、CDK2和裂解caspase-3的表达。我们发现，EGFL7和CKS2在肝细胞癌组织中均有过表达，且两者呈正相关。Egfl7基因敲除可明显抑制肝癌细胞增殖，促进细胞凋亡，同时CKS2和CDK2表达降低，caspase-3裂解表达增加，而EGFl7过表达则相反。在裸鼠体内，Egfl7的沉默也显示出肿瘤生长的减少和蛋白表达的改变，这与其在体外对肝癌细胞的影响相似。重要的是，CKS2沉默显著抑制了EGFL7诱导的肝癌细胞的增殖和蛋白表达，Wnt/β-catenin信号通路抑制剂IWR-1-Endo显著抑制了肝癌细胞中CKS2的表达。综上所述，EGFL7通过激活Wnt/β-catenin信号通路增加CKS2的表达，促进肝癌细胞增殖，抑制细胞凋亡。
Epidermal growth factor-like domain multiple 7 (EGFL7) is an important sport stimulating factor and motility related factors significantly enhanced the tumor cell metastasis and overexpressed in many cancers, including hepatocellular carcinoma (HCC), associated with tumorigenesis. However, the molecular mechanism by which EGFL7 regulates HCC cell proliferation and apoptosis and the correlation between EGFL7 and cyclin-dependent kinases regulatory subunit 2 (CKS2), which is essential for biological function, have not fully explained. In this study, EGFL7 and CKS2 expression in patients with HCC was measured by real-time polymerase chain reaction and immunohistochemistry. After HCC cells respectively transfected with pLKO.1-EGFL7-shRNA, pLVX-Puro-EGFL7 recombined vector or CKS2 small interfering RNA, cell counting kit-8 and flow cytometry was performed to examine the cell proliferation and apoptosis, respectively, and the expression of beta-catenin, CKS2, CDK2, and cleaved caspase-3 was measured by Western blot analysis. We found that EGFL7 and CKS2 were overexpressed in HCC tissues and a positive correlation was found between them. EGFL7 knockdown markedly inhibited proliferation and promoted apoptosis of HCC cells, along with decreased expression of CKS2 and CDK2, but increased cleaved caspase-3 expression, while EGFL7 overexpression showed an opposite effect. EGFL7 silencing in nude mice also showed decreased tumor growth and altered protein expression similar to its effect in HCC cells in vitro. Importantly, CKS2 silencing significantly inhibited EGFL7-induced HCC cell proliferation and protein expression, and Wnt/beta-catenin signaling pathway inhibitor IWR-1-endo significantly inhibited CKS2 expression in HCC cells. Taken together, EGFL7 promotes HCC cell proliferation and inhibits cell apoptosis through increasing CKS2 expression by activating Wnt/beta-catenin signaling.