Liang Chaojie, Xu Yingchen, Ge Hua, Xing Bingchen, Li Guanqun, Li Guangming, Wu Jixiang
Department of General Surgery, Beijing Tongren Hospital, Capital Medical University, Dongcheng, Beijing 100730, China.
Oncotarget. 2017 Nov 18;8(64):107543-107557. doi: 10.18632/oncotarget.22504. eCollection 2017 Dec 8.
Recent studies have shown that miR-564 is closely related to the development of various tumors, including breast cancer, lung cancer and glioma. However, few studies have examined miR-564 in hepatocellular carcinoma (HCC). Here, we demonstrated that miR-564 expression in HCC tissues was lower than that in adjacent noncancerous tissues and that miR-564 expression was associated with tumor size, tumor number and vein invasion. Bioinformatics analyses showed that low levels of miR-564 were correlated with poor prognosis. Furthermore, upregulation of miR-564 impaired SMCC7721 and MHCC97H cell proliferation, migration and invasion and reduced tumorigenesis . Next, we found that GRB2 was a direct target gene of miR-564 in the HCC cell lines. GRB2 was highly expressed in HCC tissues and negatively correlated with miR-564 expression levels. When GRB2 was downregulated by GRB2-siRNA, HCC cell proliferation, invasion and metastasis were impaired, and restoring GRB2 expression partially reversed the inhibitory effects of miR-564. Western blot analysis showed that miR-564 overexpression reduced GRB2 expression in HCC cell lines and inhibited ERK1/2 and AKT phosphorylation. miR-564 overexpression also upregulated the epithelial-like cell marker E-cadherin and downregulated the interstitial cell-like markers N-cadherin and vimentin. These results suggest that miR-564 inhibits the malignant phenotype of HCC cells by targeting the GRB2-ERK1/2-AKT axis. Consequently, miR-564 may be used as a prognostic marker and therapeutic target for HCC.
最近的研究表明，miR-564与包括乳腺癌、肺癌和神经胶质瘤在内的各种肿瘤的发生发展密切相关。然而，很少有研究对肝细胞癌（HCC）中的miR-564进行检测。在此，我们证明HCC组织中miR-564的表达低于相邻的非癌组织，且miR-564的表达与肿瘤大小、肿瘤数量和静脉侵犯相关。生物信息学分析表明，低水平的miR-564与预后不良相关。此外，miR-564的上调损害了SMCC7721和MHCC97H细胞的增殖、迁移和侵袭，并减少了肿瘤发生。接下来，我们发现GRB2是HCC细胞系中miR-564的直接靶基因。GRB2在HCC组织中高表达，且与miR-５64表达水平呈负相关。当通过GRB2-siRNA下调GRB2时，HCC细胞的增殖、侵袭和转移受到损害，恢复GRB2表达部分逆转了miR-564的抑制作用。蛋白质印迹分析表明，miR-564过表达降低了HCC细胞系中GRB2的表达，并抑制了ERK1/2和AKT的磷酸化。miR-564过表达还上调了上皮样细胞标志物E-钙黏蛋白，并下调了间质样细胞标志物N-钙黏蛋白和波形蛋白。这些结果表明，miR-564通过靶向GRB2-ERK1/2-AKT轴抑制HCC细胞的恶性表型。因此，miR-564可能用作HCC的预后标志物和治疗靶点。
