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1.兰州大学第一临床医学院,兰州 730030
2.兰州大学第一医院普外科六病区,兰州 730030
陈 燕,Email:qinshaojiec@163.com,研究方向:恶性肿瘤侵袭转移分子机制研究。
姜 雷,Email:jiangzx@lzu.edu.cn。
收稿:2023-08-18,
修回:2023-08-20,
录用:2023-09-30,
纸质出版:2023-10-28
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陈燕,姜雷,闵光涛,等.miR-149-5p通过靶向AEBP1调控胃癌细胞的迁移侵袭[J].重庆医科大学学报,2023,48(10):1159-1165.
Chen Yan,Jiang Lei,Min Guangtao,et al.miR-149-5p targets AEBP1 to inhibit migration and invasion of gastric cancer cells[J].Journal of Chongqing Medical University,2023,48(10):1159-1165.
陈燕,姜雷,闵光涛,等.miR-149-5p通过靶向AEBP1调控胃癌细胞的迁移侵袭[J].重庆医科大学学报,2023,48(10):1159-1165. DOI: 10.13406/j.cnki.cyxb.003346.
Chen Yan,Jiang Lei,Min Guangtao,et al.miR-149-5p targets AEBP1 to inhibit migration and invasion of gastric cancer cells[J].Journal of Chongqing Medical University,2023,48(10):1159-1165. DOI: 10.13406/j.cnki.cyxb.003346.
目的
2
探讨miR-149-5p对胃癌细胞迁移侵袭能力的影响及分子机制。
方法
2
qRT-PCR检测胃癌细胞株和组织标本中miR-149-5p的表达,分析其表达水平与胃癌患者临床病理特征参数及预后的相关性,建立过表达和干扰miR-149-5p的胃癌细胞株,Transwell实验检测miR-149-5p表达水平对胃癌癌细胞迁移侵袭能力的影响;生物信息学网站预测miR-149-5p的靶基因,采用荧光素酶报告基因实验加以验证。
结果
2
miR-149-5p在胃癌组织和细胞株中均低表达(均
P
<
0.05)。miR-149-5p的表达水平与胃癌患者的浸润深度(
P
=0.016)、淋巴结转移(
P
=0.001) 和TNM分期(
P
=0.023) 相关。miR-149-5p低表达是胃癌患者总生存期的独立危险因素。与对照组相比,miR-149-5p mimics明显抑制胃癌细胞的迁移侵袭能力(均
P
<
0.01),而转染miR-149-5p inhibitors后得到了相反的结果(均
P
<
0.05)。miR-149-5p靶向调控脂肪细胞增强结合蛋白1(Adipocyte enhancer binding protein 1,AEBP1)蛋白的表达。荧光素酶报告基因实验表明,miR-149-5p明显抑制了野生型AEBP1载体的荧光素酶活性(
P
<
0.01),而突变型的荧光素酶活性不受影响。敲低AEBP1可部分逆转下调miR-149-5p对胃癌细胞迁移侵袭能力的影响。
结论
2
miR-149-5p在胃癌组织中低表达,通过靶向AEBP1负性调控胃癌癌细胞的迁移侵袭。
Objective
2
To investigate the effects of miR-149-5p in regulating the migration and invasion ability of gastric cancer(GC) cells and the molecular mechanism.
Methods
2
The expression of miR-149-5p in GC cell lines and tissue specimens was measured by qRT-PCR to analyze the relationship between miR-149-5p expression and the clinicopathological parameters and prognosis of patients with GC. We separately overexpressed and knocked down miR-149-5p in GC cells to investigate the expression level of miR-149-5p on the migration and invasion ability of GC cells using Transwell assay. The target genes of miR-149-5p were predicted using bioinformatic tools,which were verified using luciferase reporter gene assay.
Results
2
The expression of miR-149-5p was significantly down-regulated in both GC tissues and cell lines(
P
<
0.05). The expression level of miR-149-5p was significantly correlated with
the depth of invasion(
P
=0.016),lymph node metastasis(
P
=0.001),and TNM stage(
P
=0.023) of patients with GC. Low expression of miR-149-5p was an independent risk factor for the overall survival of patients with GC. Compared with the control group,miR-149-5p mimics significantly inhibited the migration and invasion ability of gastric cancer cells(
P
<
0.01),while transfection with miR-149-5p inhibitors produced the opposite effects(
P
<
0.05). miR-149-5p targeted the expression of adipocyte enhancer-binding protein 1(AEBP1). The luciferase reporter gene assay showed that miR-149-5p significantly inhibited the luciferase activity of the wild-type AEBP1 vector(
P
<
0.01),with no effects on the luciferase activity of the mutant type. Knockdown of AEBP1 could partly reverse the effects of downregulating miR-149-5p on the migration and invasion ability of GC cells.
Conclusion
2
miR-149-5p is lowly expressed in GC tissues,which can negatively regulate the migration and invasion of GC cells by targeting AEBP1 protein expression.
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