文献类型：期刊文献

中文题名：微RNA-let-7a-5p通过调节鼠双微体2/肿瘤蛋白53信号通路抑制胃癌细胞增殖

英文题名：MicroRNA-let-7a-5p suppressing gastric cancer cell proliferation by regulating the mouse double minute 2/tumor protein 53 signaling pathway

作者：黄勇[1,4];赵星星[2];程安琪[2];苏韫[1,3];张晗[1,3];王芳[5];龚红霞[1,3];曹旺杰[1,3];刘永琦[1,3]

第一作者：黄勇

机构：[1]甘肃中医药大学基础医学院,兰州730000;[2]甘肃中医药大学第一临床医学院,兰州730000;[3]甘肃中医药大学甘肃省高校重大疾病分子医学与中医药防治研究省级重点实验室,兰州730000;[4]甘肃中医药大学病理教研室,兰州730000;[5]甘肃中医药大学公共卫生学院检验与检疫教研室,兰州730000

第一机构：甘肃中医药大学基础医学院（敦煌医学研究所）

年份：2026

卷号：57

期号：2

起止页码：218

中文期刊名：解剖学报

外文期刊名：Acta Anatomica Sinica

基金：甘肃省自然科学基金(25JRRA256);国家卫生健康委员会胃肠肿瘤诊治重点实验室开放课题(23GSSYA-16);甘肃中医药大学科学与研究创新基金重点项目(2023KCZD-3);甘肃省中医药防治慢性疾病重点实验室开放课题(GSSYLXM-05);甘肃省高校中(藏)药化学与质量研究省级重点实验室开放基金(zzy-2022-06)。

语种：中文

中文关键词：胃癌;微RNA-let-7a-5p;鼠双微体2/肿瘤蛋白53信号通路;周期阻滞;免疫印迹法;人

外文关键词：Gastric cancer;MicroRNA-let-7a-5p;Mouse double minute 2;tumor protein 53 signaling pathway;Cycle arrest;Western blotting;Human

摘要：目的探讨微RNA let-7a-5p通过调控鼠双微体2(MDM2)/肿瘤蛋白53(p53)信号通路对胃癌细胞凋亡和周期阻滞的作用。方法收集20对胃癌组织及配对的癌旁组织标本,检测let-7a-5p表达;生物信息学数据库分析let-7a-5p在胃癌中的表达及与预后的关系,预测let-7a-5p的靶基因;构建let-7a-5p过表达和低表达细胞模型;CCK-8、划痕实验、流式细胞术分别检测细胞增殖、迁移、凋亡和周期分布;双荧光素酶报告基因实验验证let-7a-5p与MDM2的靶向关系;Western blotting和Real-time PCR检测MDM2、p53及通路中细胞周期蛋白D1(Cyclin D1)、细胞髓细胞增生原癌基因(c-myc)、Bcl-2和Bax的表达。结果与癌旁组织相比,let-7a-5p在胃癌组织中低表达(P<0.05),且低表达患者预后较差;转染let-7a-5p模拟物(let-7a-5p mimics)可明显过表达人胃癌细胞HGC-27细胞中let-7a-5p的水平(P<0.01),而转染let-7a-5p抑制剂(let-7a-5p inhibitor)可降低人胃癌细胞AGS中let-7a-5p的表达(P<0.05)。Let-7a-5p过表达可抑制HGC-27的增殖和迁移,促进凋亡和周期阻滞;Let-7a-5p低表达则促进AGS细胞增殖和迁移,抑制凋亡和周期阻滞;Targeiscan数据库和双荧光素酶报告基因实验结果表明,let-7a-5p直接靶向MDM2,负调控其表达;Western blotting和Real-time PCR实验结果表明,let-7a-5p过表达可上调p53和Bax的表达,下调MDM2、Bcl-2、Cyclin D1和c-myc的表达。Let-7a-5p低表达可上调MDM2、Bcl-2、Cyclin D1、c-myc的表达,下调p53、Bax的表达。结论Let-7a-5p通过抑制MDM2/p53信号通路促进胃癌细胞凋亡和周期阻滞。
Objective To investigate the role and mechanism of microRNA-let-7a-5p(miRAN-let-7a-5p)in regulating apoptosis and cell cycle arrest in gastric cancer cells via the mouse double minute 2(MDM2)/tumor protein 53(p53)signaling pathway.Methods Twenty pairs of gastric cancer tissues and matched adjacent non-tumor tissues were collected to detect let-7a-5p expression.Bioinformatics databases were used to analyze the expression of let-7a-5p in gastric cancer and its correlation with patient prognosis.Potential target genes of let-7a-5p were predicted;Both overexpression and knockdown cell models of let-7a-5p were established.Cell proliferation,migration,apoptosis,and cell cycle distribution were assessed using the CCK-8 assay,wound healing assay,and flow cytometry,respectively.The targeting relationship between let-7a-5p and MDM2 was verified by a dual-luciferase reporter assay.Western blotting and Real-time PCR were performed to measure the expression of MDM2,p53,and downstream factors,including Cyclin D1,cellular myelocytomatosis oncogene product(c-myc)、tumor protein 53(p53),Bcl-2 and Bax.Results Real-time PCR results showed that let-7a-5p was significantly downregulated in gastric cancer tissues compared with adjacent normal tissues(P<0.05),and low let-7a-5p expression was associated with poor prognosis.Transfection with let-7a-5p mimics markedly increased let-7a-5p levels in human gastric cancer cells HGC-27(P<0.01),while let-7a-5p inhibitor reduced its expression in human gastric cancer cells AGS(P<0.05).Overexpression of let-7a-5p inhibited proliferation and migration,promoting apoptosis and cell cycle arrest in HGC-27 cells.In contrast,knockdown of let-7a-5p enhanced proliferation and migration,suppressed apoptosis and cell cycle arrest in AGS cells.Target prediction and dual-luciferase reporter assays confirmed that let-7a-5p directly targeted MDM2 and negatively regulated its expression.Western blotting and Real-time PCR results demonstrated that let-7a-5p overexpression upregulated p53 and Bax,and downregulated MDM2,Bcl-2,CyclinD1,and c-myc.Conversely,let-7a-5p knockdown increased the expression of MDM2,Bcl-2,Cyclin D1,and c-myc,and decreased p53 and Bax levels.Conclusion Let-7a-5p promotes apoptosis and cell cycle arrest in gastric cancer cells by suppressing the MDM2/p53 signaling pathway.