miR-199a-3p 通过靶向 ETNK1 促进胃癌细胞的侵袭和迁移，并与不良预后相关。

miR-199a-3p targets ETNK1 to promote invasion and migration in gastric cancer cells and is associated with poor prognosis.

机构信息

Clinical Research Institute, Zhejiang Provincial People's Hospital, People's Hospital of Hangzhou Medical College, Hangzhou, 310014, Zhejiang, China; Key Laboratory of Gastroenterology of Zhejiang Province, Hangzhou, 310014, PR China.

Departments of Gastrointestinal and Pancreatic Surgery, Zhejiang Provincial People's Hospital, People's Hospital of Hangzhou Medical College, Hangzhou, 310014, PR China; Key Laboratory of Gastroenterology of Zhejiang Province, Hangzhou, 310014, PR China.

出版信息

Pathol Res Pract. 2019 Sep;215(9):152511. doi: 10.1016/j.prp.2019.152511. Epub 2019 Jun 21.

DOI:10.1016/j.prp.2019.152511

PMID:31255331

Abstract

PURPOSE

To investigate the prognostic significance of miR-199a-3p and its role in invasion and metastasis in gastric cancer.

METHODS

miR-199a-3p expression in 436 formalin-fixed and 39 frozen gastric cancer tissues was investigated by in situ hybridization and RT-PCR, respectively. The role of miR-199a-3p in the migration and invasion of gastric cancer cells was determined in overexpression and inhibitor studies using transwell assays and the SGC-7901, BGC-823 and MGC-803 gastric cancer cells lines. The effect of miR-199a-3p expression on ethanolamine kinase 1 (ETNK1) levels was determined by western botting.

RESULTS

miR-199a-3p was significantly up-regulated in AGS, SGC-7901, BGC-823 and MGC-803 gastric cancer cells, when compared with GES-1 non-malignant gastric epithelial cells. In situ hybridization studies revealed that human non-tumor gastric mucosa samples were negative for miR-199a-3p expression, while 162 of 436 (37.16%) cases of gastric cancer demonstrated positive expression. miR-199a-3p overexpression was associated with tumor size, Lauren classification, depth of invasion, lymph node and distant metastasis, TNM stage and prognosis. In patients with I, II and III stage tumors, high miR-199a-3p expression was associated with a significantly lower 5-year survival rate. miR-199a-3p overexpression was associated with increased cell migration and invasion. ETNK1 expression was inhibited following miR-199a-3p overexpression in BGC-823 and SGC-7901 cells, and elevated following miR-199a-3p suppression in MGC-803 cells.

CONCLUSION

miR-199a-3p is highly expressed in gastric cancer, and correlates with invasion, metastasis and prognosis. miR-199a-3p regulates the invasion and migration of gastric cancer cells by targeting ETNK1. Consequently, miR-199a-3p may serve as a prognostic indicator in gastric cancer.

摘要

目的

研究 miR-199a-3p 的预后意义及其在胃癌侵袭转移中的作用。

方法

采用原位杂交和 RT-PCR 分别检测 436 例福尔马林固定和 39 例冷冻胃癌组织中 miR-199a-3p 的表达。通过转染实验和体外侵袭实验，在过表达和抑制研究中使用 SGC-7901、BGC-823 和 MGC-803 胃癌细胞系，研究 miR-199a-3p 在胃癌细胞迁移和侵袭中的作用。通过 Western blot 确定 miR-199a-3p 表达对乙醇胺激酶 1 (ETNK1) 水平的影响。

结果

与 GES-1 非恶性胃上皮细胞相比，AGS、SGC-7901、BGC-823 和 MGC-803 胃癌细胞中 miR-199a-3p 表达明显上调。原位杂交研究显示，人非肿瘤性胃黏膜标本无 miR-199a-3p 表达，而 436 例胃癌病例中有 162 例（37.16%）呈阳性表达。miR-199a-3p 过表达与肿瘤大小、Lauren 分类、浸润深度、淋巴结和远处转移、TNM 分期和预后有关。在 I、II 和 III 期肿瘤患者中，高 miR-199a-3p 表达与 5 年生存率显著降低相关。miR-199a-3p 过表达与细胞迁移和侵袭增加相关。BGC-823 和 SGC-7901 细胞中 miR-199a-3p 过表达抑制 ETNK1 表达，而 MGC-803 细胞中 miR-199a-3p 抑制后 ETNK1 表达升高。