目的：探讨miR-153-3p对胃癌SGC7901细胞增殖、侵袭和迁移的作用及其机制。方法：收集2018年5月至2020年6 月宁夏医科大学总医院收治的60例胃癌患者的癌和配对癌旁组织标本，以及人胃癌细胞系NCI-N87、AGS、SNU-5、SGC7901和胃上 皮细胞 GES-1，qPCR 法检测 miR-153-3p 在胃癌组织与细胞中的表达水平。将 miR-153-3p mimic 及 mimic 对照序列转染至 SGC7901细胞，用CCK-8、克隆形成、流式细胞术、TUNEL、Transwell和划痕愈合实验分别检测上调miR-153-3p对SGC7901细胞 增殖、凋亡、侵袭和迁移的影响。构建裸鼠SGC7901细胞移植瘤模型，观察miR-153-3p对肿瘤生长的影响。通过生物信息学数 据库和双荧光素酶报告基因实验预测并验证miR-153-3p与FZD3靶向关系，WB法检测miR-153-3p对FZD3蛋白及Wnt/β-catenin 通路相关蛋白表达的影响。结果：miR-153-3p在胃癌组织和细胞中表达水平分别显著低于癌旁组织和GES-1细胞（均P<0.01），以SGC7901细胞中表达水平最低。上调 miR-153-3p 显著抑制 SGC7901 细胞的增殖、侵袭和迁移能力，并提高细胞凋亡率 （均P<0.01），同时上调细胞中E-cadherin表达而下调N-cadherin、MMP2和MMP9表达（均P<0.01）。在体内实验表明，静脉注射 miR-153-3p mimic显著降低移植瘤体积和瘤组织中Ki-67表达而上调P57表达（均P<0.01）。机制分析表明 ，miR-153-3p 靶向 结合FZD3基因的3′UTR区域，上调 miR-153-3p 会抑制FZD3表达并上调β-catenin、TCF-4和cyclin D1水平（均P<0.01）。结论： miR-153-3p靶向FZD3并通过Wnt/β-catenin信号通路调控胃癌SGC7901细胞的增殖、侵袭和迁移。

Objective: To investigate the effect of miR-153-3p on the proliferation, invasion and migration of gastric cancer SGC7901 cells and its molecular mechanism. Methods: A total of 60 pairs of cancer tissues and corresponding para-cancerous tissues of gastric cancer patients who were treated in the General Hospital of Ningxia Medical University during May 2018 and June 2020 were collected for this study; in addition, human gastric cancer cell lines (NCI-N87, AGS, SNU-5, SGC7901) and normal gastric epithelial GES-1 cells were also collected. The expression level of miR-153-3p in gastric cancer tissues and cells was detected by qRT-PCR. miR-153-3p mimics and mimic control were transfected into SGC7901 cells, respectively. The effects of miR-153-3p overexpression on proliferation, apoptosis, invasion and migration of gastric cancer SGC7901 cells were respectively determined using CCK-8, Clone formation assay, Flow cytometry, TUNEL, Transwell and Wound-healing assay. SGC7901 cell transplanted xenograft tumor model in nude mice was established to explore the effect of miR-153-3p on tumor growth. The targeting relationship between miR-153-3p and FZD3 was predicted and verified by bioinformatics database and Dual-luciferase reporter gene assay, respectively. The effects of miR-153-3p on expression of FZD3 protein and Wnt/β-catenin pathway related proteins were determined by Western blotting. Results: The expression level of miR-153-3p in gastric cancer tissues and cells was significantly lower than that in para-cancerous tissues and gastric epithelial cells, with the lowest expression in SGC7901 cells (all P<0.01). Up-regulation of miR-153-3p significantly inhibited the proliferation, invasion and migration abilities of SGC7901 cells, and increased the rate of apoptotic cells (all P<0.01). In addition, up-regulation of miR-153-3p significantly promoted the expression of E-cadherin but significantly suppressed the expression of N-cadherin, matrix metalloproteinase-2 (MMP2) and matrix metalloproteinase-9 (MMP9) (all P<0.01). In vivo experiments showed that intravenous injection of miR-153-3p mimics significantly reduced tumor size and Ki-67 expression, but significantly increased the expression of P57 in tumor tissues (all P<0.01). The mechanism analysis showed that miR-153-3p could bind to the 3'UTR region of FZD3 mRNA, and up-regulation of miR-153-3p inhibited the expression of FZD3 and increased the levels of β -catenin, TCF-4 and cyclin D1 (all P<0.01). Conclusion: MiR-153-3p regulates the proliferation, invasion and migration of gastric cancer SGC7901 cells via targeting FZD3 gene and regulating Wnt/β-catenin signaling pathway.

宁夏自然科学基金资助项目（No.2019AAC03200, No.2019AAC03225）