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[微小RNA-96-5p通过靶向FoxQ1抑制胃癌细胞的增殖和迁移]

[miRNA-96-5p inhibits the proliferation and migration of gastric cancer cells by targeting FoxQ1].

作者信息

Yang X Y, Li N, Deng W Y, Ma Y J, Han X L, Zhang Z Y, Xie J L, Luo S X

机构信息

Department of Pharmacy, Affiliated Cancer Hospital of Zhengzhou University, Zhengzhou 450008, China.

Department of Gastroenterology, Affiliated Cancer Hospital of Zhengzhou University, Zhengzhou 450008, China.

出版信息

Zhonghua Zhong Liu Za Zhi. 2019 Mar 23;41(3):193-199. doi: 10.3760/cma.j.issn.0253-3766.2019.03.008.

Abstract

To investigate the role of microRNA-96-5p in the proliferation and invasion of gastric cancer cells and its molecular mechanism. From June 2015 to January 2017, 53 resected specimens were collected. The transcriptional levels of microRNA-96-5p and forkhead box Q1 (FoxQ1) in gastric cancer tissues and the matched para-cancerous tissues were quantified by quantitative real-time PCR (qRT-PCR). The expression of FoxQ1 protein was also detected by immunohistochemistry (IHC). The relationship between microRNA-96-5p expression and the clinicopathological features of gastric cancer and its correlation with FoxQ1 expression were analyzed. The expressions of miRNA-96-5p in gastric cancer tissue and adjacent normal tissue were detected by qRT-PCR. miRNA-96-5p mimics was transfected to BGC-823 gastric cancer cells. The effects of miRNA-96-5p on cell proliferation and invasion were detected by cell counting kit-8 (CCK-8) assay and Transwell assay, respectively. The protein expressions of FoxQ1, E-cadherin and vimentin were determined by western blot. The relationship between FoxQ1 and miRNA-96-5p expressed in BGC-823 cells was detected by dual-luciferase reporter assay. The median expression of miRNA-96-5p in gastric cancer tissue was 1.05, significantly lower than 3.23 of para-cancerous tissues (<0.05). The positive rate of FoxQ1 expression in gastric cancer tissue was 71.7%, significantly higher than 28.3% of para-cancerous tissues (<0.05). The expression of FoxQ1 was negatively corelated with the level of miRNA-96-5p (=-0.613, =0.006). The expression of miRNA-96-5p in gastric cancer cell BGC-823 was significantly decreased compared with normal gastric epithelial cell (0.96±0.08 vs 2.84±0.15, <0.05). The results of CCK-8 assay and Transwell assay showed that overexpression of miRNA-96-5p significantly reduced the proliferation and invasion abilities of gastric cancer cells (<0.05). Overexpression of miRNA-96-5p decreased the protein level of FoxQ1. Moreover, it upregulated the expression of E-cadherin and downregulated the expression of vimentin. The result of dual-luciferase-3'-UTR reporter assay confirmed that miRNA-96-5p binds to the 3'UTR of FoxQ1. miRNA-96-5p may suppress the proliferation, migration and epithelial-mesenchymal transition (EMT) of gastric cancer cell by down-regulation of FoxQ1.

摘要

探讨微小RNA-96-5p在胃癌细胞增殖和侵袭中的作用及其分子机制。2015年6月至2017年1月,收集53例手术切除标本。采用定量实时荧光定量PCR(qRT-PCR)检测胃癌组织及配对癌旁组织中微小RNA-96-5p和叉头框Q1(FoxQ1)的转录水平。采用免疫组织化学(IHC)检测FoxQ1蛋白表达。分析微小RNA-96-5p表达与胃癌临床病理特征的关系及其与FoxQ1表达的相关性。采用qRT-PCR检测胃癌组织及癌旁正常组织中miRNA-96-5p的表达。将miRNA-96-5p模拟物转染至BGC-823胃癌细胞。分别采用细胞计数试剂盒-8(CCK-8)法和Transwell法检测miRNA-96-5p对细胞增殖和侵袭的影响。采用蛋白质印迹法检测FoxQ1、E-钙黏蛋白和波形蛋白的蛋白表达。采用双荧光素酶报告基因检测法检测BGC-823细胞中FoxQ1与miRNA-96-5p的关系。胃癌组织中miRNA-96-5p的中位表达量为1.05,显著低于癌旁组织的3.23(<0.05)。胃癌组织中FoxQ1表达的阳性率为71.7%,显著高于癌旁组织的28.3%(<0.05)。FoxQ1的表达与miRNA-96-5p水平呈负相关(r=-0.613,P=0.006)。与正常胃上皮细胞相比,胃癌细胞BGC-823中miRNA-96-5p的表达显著降低(0.96±0.08 vs 2.84±0.15,<0.05)。CCK-8法和Transwell法结果显示,miRNA-96-5p过表达显著降低胃癌细胞的增殖和侵袭能力(<0.05)。miRNA-96-5p过表达降低了FoxQ1的蛋白水平。此外,它上调了E-钙黏蛋白的表达,下调了波形蛋白的表达。双荧光素酶-3'-UTR报告基因检测结果证实miRNA-96-5p与FoxQ1的3'UTR结合。miRNA-96-5p可能通过下调FoxQ1抑制胃癌细胞的增殖、迁移和上皮-间质转化(EMT)。

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