miR-501通过下调eIF4E3促进胃癌细胞增殖、迁移和侵袭  

miR-501 promotes gastric cancer cell proliferation,migration and invasion through targeting eIF4E3

作　　者：黄金丽 关宏伟[2] 卢颖 侯力[1] 黄玉红[1] 张晴晴[1] 宋波[1] HUANG Jin-li;GUAN Hong-wei;LU Ying;HOU Li;HUANG Yu-hong;ZHANG Qing-qing;SONG Bo(Department of Pathology and Forensics,Dalian Medical University,Dalian 116044,China;Department of Pathology,First Affiliated Hospital of Dalian Medical University,Dalian 116011,China;Teaching Laboratory of Morphology,Dalian Medical University,Dalian 116044,China)

机构地区：[1]大连医科大学病理学与法医学教研室,大连116044 [2]大连医科大学附属第一医院病理科,大连116011 [3]大连医科大学形态学实验室,大连116044

出　　处：《临床与实验病理学杂志》2022年第8期897-902,共6页Chinese Journal of Clinical and Experimental Pathology

基　　金：辽宁省教育厅基本科研项目(LZ2019035)。

摘　　要：目的 探讨miR-501对胃癌细胞增殖、迁移和侵袭能力的影响及分子机制。方法 采用qRT-PCR检测miR-501和eIF4E3 mRNA表达,应用Western blot法检测eIF4E3蛋白水平,CCK-8和细胞克隆形成实验检测细胞增殖能力,运用Transwell小室实验检测细胞迁移和侵袭能力,双荧光素酶报告基因实验检测miR-501与eIF4E3的靶向关系。结果 miR-501在胃癌细胞系SGC-7901、BGC-823和MGC-803中表达水平分别是正常胃黏膜上皮细胞GES-1的2.78±0.28、2.51±0.30和1.62±0.14倍(P均<0.05)。TCGA数据库分析显示miR-501在胃癌组织中表达显著高于正常胃组织(P<0.001)。miR-501 mimic转染BGC-823细胞(miR-501组)增殖活性明显高于miRNA无关序列对照(NC组)(P均<0.05);Transwell小室迁移实验miR-501组穿膜细胞数(587±16)个,高于阴性NC组的(396±34)个(P<0.05);Transwell小室侵袭实验miR-501组穿膜细胞数(553±43)个,高于NC组的(398±21)个(P<0.05)。miR-501和野生型eIF4E3载体共转染细胞荧光素酶活性明显降低(P<0.01)。转染miR-501可使eIF4E3 mRNA和蛋白水平显著降低。与siRNA无关序列(NCi组)相比,eIF4E3敲低(eIF4E3-si组)可显著增强BGC-823细胞增殖活性(P<0.05);Transwell小室迁移实验eIF4E3-si组穿膜细胞数(299±32)个,多于阴性NCi组的(182±13)个(P<0.01),Transwell小室侵袭实验也呈相同趋势(P<0.01)。TCGA数据库分析显示eIF4E3 mRNA在正常胃组织中表达显著高于胃癌组织(P<0.001)。结论 miR-501可通过靶向下调抑癌基因eIF4E3表达,促进胃癌细胞增殖、迁移和侵袭能力;miR-501有望成为胃癌治疗的潜在靶点。Purpose To investigate the effects of miR-501 on gastric cancer cell proliferation,migration and invasion and the molecular mechanism.Methods The expression of miR-501 and eIF4 E3 mRNA was detected by qRT-PCR.The eIF4 E3 protein expression was measured by Western blot.Cell growth was determined by CCK-8 and cell colony formation assays,and cell migratory and invasive capacities were detected by Transwell chamber assays.The Dual-luciferase reporter assay was used to evaluate the direct binding between miR-501 and eIF4 E3.Results The expression levels of miR-501 in gastric cancer cell lines SGC-7901,BGC-823 and MGC-803 were 2.78±0.28,2.51±0.30 and 1.62±0.14 times higher than those in normal gastric mucosa epithelial cells GES-1 respectively(P<0.05).TCGA database analysis showed that the expression of miR-501 in gastric cancer tissues was significantly higher than that in normal gastric tissues(P<0.001).BGC-823 cells transfected with miR-501 mimic(miR-501 group) showed significantly higher proliferative potential than the non-specific miRNA control(NC group)(P<0.05).Transwell chamber migration assay showed that the number of cells penetrating through the membrane in miR-501 group(587±16) was significantly higher than that in negative NC group(396±34)(P<0.05),and Transwell chamber invasion assay showed that the number of cells penetrating through the membrane in miR-501 group(553±43) was significantly higher than that in NC group(398±21)(P<0.05).Co-transfection of miR-501 and wild type eIF4 E3 vector dramatically reduced the luciferase activity(P<0.01).Ectopic transfection of miR-501 markedly reduced the levels of mRNA and protein of eIF4 E3.Silenced eIF4 E3 expression(eIF4 E3-si group) significantly improved the potential of BGC-823 cell growth compared with the non-specific siRNA control(NCi group)(P<0.05).Transwell chamber migration assay showed that the number of cells penetrating through the membrane in eIF4 E3-si group(299±32) was significantly higher than that in negative NCi group(182±13)(P<0.01),and

关 键 词：胃肿瘤 miR-501 eIF4E3 增殖 迁移 侵袭 

分 类 号：R735.2[医药卫生—肿瘤]