MicroRNA-9通过NRP1抑制胃癌SGC-7901细胞上皮-间充质转化功能

发布时间：2018-06-20 12:10

本文选题：胃癌 + 微小RNA-　；参考：《中国病理生理杂志》2017年07期

【摘要】：目的:研究微小RNA-9(microRNA-9,miR-9)对胃癌SGC-7901细胞上皮-间充质转化(EMT)功能的影响及其相关机制。方法:SGC-7901胃癌细胞株分别转染miR-9 mimics和阴性对照序列(negative control mimic,NCM),作为miR-9组和NCM组,并设立未转染对照(control)组,采用RT-qPCR法检测各组细胞miR-9的含量,Transwell实验检测3组细胞迁移能力和侵袭能力,Western blot法检测3组细胞的N-cadherin、E-cadherin、α-catenin和神经纤毛蛋白1(NRP1)表达水平。采用Western blot法检测NRP1过表达对miR-9抑制EMT的拮抗作用。双萤光素酶实验检测miR-9与NRP1的关系。结果:miR-9组的miR-9表达水平明显上调,为control组的538倍(P0.05)。miR-9组的迁移细胞数量明显低于control组(P0.05)。miR-9组的侵袭细胞数量明显低于control组(P0.05)。miR-9组细胞的N-cadherin和NRP1蛋白表达量明显降低,E-cadherin及α-catenin蛋白表达量明显升高。而NRP1及miR-9均过表达组胃癌细胞中N-cadherin蛋白表达量明显升高,E-cadherin及α-catenin蛋白表达量明显降低。双萤光素酶检验结果显示NRP1为miR-9的下游靶基因(P0.05)。结论:miR-9可能通过降低下游靶基因NRP1水平影响EMT相关蛋白表达,抑制胃癌SGC-7901细胞的EMT功能。
[Abstract]:Aim: to study the effect of microRNA-9 miR-9 on the epithelial-mesenchymal transformation (EMTT) function of gastric cancer SGC-7901 cells and its related mechanism. Methods: human SGC-7901 gastric cancer cell line was transfected with miR-9 mimics and negative control mimicia NCMI respectively as miR-9 group and NCM-negative control group. The content of miR-9 was detected by RT-qPCR and the expression levels of N-cadherin-E-cadherin, 伪 -catenin and neurociliated protein 1nRP1 were detected by Transwell assay and Western blot assay. Western blot assay was used to detect the antagonistic effect of NRP1 overexpression on the inhibition of EMT by miR-9. The relationship between miR-9 and NRP1 was detected by double luciferase assay. Results the miR-9 expression level was significantly up-regulated in the miR-9 group. The number of migration cells in control group was significantly lower than that in control group. The number of invasive cells was significantly lower in control group than that in control group. The expression of N-cadherin and NRP1 protein in control group was significantly lower than that in control group and the expression of E-cadherin and 伪 -catenin protein was significantly increased. The expression of N-cadherin and 伪 -catenin in gastric cancer cells were significantly increased in both NRP1 and miR-9 overexpression groups, and the expression of E-cadherin and 伪 -catenin were significantly decreased. The results of double luciferase assay showed that NRP1 was the downstream target gene of miR-9. Conclusion the expression of EMT related protein may be affected by reducing the level of downstream target gene NRP1 and inhibiting the EMT function of gastric cancer SGC-7901 cells.
【作者单位】：天津市第五中心医院消化科;
【基金】：滨海新区卫生局基金资助项目(No.2013BWKY031)
【分类号】：R735.2

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