山东医药2024,Vol.64Issue(5) :22-25.DOI:10.3969/j.issn.1002-266X.2024.05.005

miR-203a-3p对食管癌细胞侵袭迁移能力的影响及其与Survivin的靶向调控关系

Effects of miR-203a-3p on invasion and migration of esophageal cancer cells and its targeted regulatory relationship with Survivin

郑竞雄 黄景涛 孙光蕊 赵宝山 梁宗英
山东医药2024,Vol.64Issue(5) :22-25.DOI:10.3969/j.issn.1002-266X.2024.05.005

miR-203a-3p对食管癌细胞侵袭迁移能力的影响及其与Survivin的靶向调控关系

Effects of miR-203a-3p on invasion and migration of esophageal cancer cells and its targeted regulatory relationship with Survivin

郑竞雄 1黄景涛 1孙光蕊 1赵宝山 1梁宗英1
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作者信息

  • 1. 承德医学院附属医院胸外科,河北承德 067000
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摘要

目的 观察微小RNA(miR)-203a-3p对食管癌细胞侵袭、迁移能力的影响并分析其与Survivin的靶向调控关系.方法 收集对数生长期的食管癌细胞TE-1及正常食管黏膜上皮细胞HEEC,RT-qPCR法检测细胞miR-203a-3p、Survivin mRNA.双荧光素酶报告基因检测法分析miR-203a-3p与Survivin的靶向调控关系.将TE-1细胞分为对照组、miR-203a-3p模拟组、miR-203a-3p抑制组及阴性对照组.miR-203a-3p模拟组转染miR-203a-3p mimic,miR-203a-3p抑制组转染miR-203a-3p inhibitor,阴性对照组转染miR-203a-3p NC,对照组不进行转染,RT-qPCR法验证转染效率.采用MTT法检测转染后2、12、24、36、48 h的细胞活力,细胞划痕实验检测细胞迁移能力,Transwell侵袭实验检测细胞侵袭能力.结果 食管癌细胞中miR-203a-3p mRNA表达低于正常食管黏膜上皮细胞,Survivin mRNA表达高于正常食管黏膜上皮细胞(P均<0.05).TargetScan数据库分析显示,Survivin基因在3′UTR端存在7个连续的可以与miR-203a-3p互补的核苷酸序列;双荧光素酶报告基因实验结果显示,共转染Survivin-WT和miR-203a-3p mimic质粒的TE-1细胞荧光素酶活性低于其他细胞(P<0.05).培养12、24、36、48 h时细胞活力miR-203a-3p抑制组>对照组、阴性对照组>miR-203a-3p模拟组(P均<0.05);细胞迁移距离及侵袭细胞数miR-203a-3p抑制组>对照组、阴性对照组>miR-203a-3p模拟组(P均<0.05).结论 miR-203a-3p高表达可抑制食管癌细胞的迁移和侵袭能力,其机制可能与靶向负调控Survivin有关.

Abstract

Objective To explore the effects of miR-203a-3p on the invasion and migration of esophageal cancer cells and to analyze the targeted regulatory relationship with Survivin.Methods Esophageal cancer TE-1 cells and nor-mal esophageal mucosal epithelial HEEC cells in logarithmic growth phase were collected.The miR-203a-3p and Survivin mRNA were tested by RT-qPCR.The targeted regulatory relationship between miR-203a-3p and Survivin was analyzed by dual luciferase reporter assay.The TE-1 cells were divided into the control group,miR-203a-3p simulation group,miR-203a-3p inhibitory group,and negative control group,respectively.Cells in the miR-203a-3p simulation group were trans-fected with miR-203a-3p mimic,miR-203a-3p-inhibited group with miR-203a-3p inhibitor,and negative control group with miR-203a-3p NC,while cells in the control group were not transfected.Transfection efficiency was verified by RT-qP-CR.Cell viability was measured by MTT at 2,12,24,36 and 48 h after transfection,cell migration was measured by Scratch assay and cell invasion was measured by Transwell invasion assay.Results The mRNA expression of miR-203a-3p was lower and the mRNA expression of Survivin was higher in the esophageal cancer cells than in the normal esophageal mucosal epithelial cells(both P<0.05).TargetScan database analysis showed that the Survivin gene had seven consecu-tive nucleotide sequences at the 3′UTR end that could be complementary to miR-203a-3p.The result of dual luciferase re-porter assay showed that TE-1 cells co-transfected with Survivin-WT and plasmid-miR-203a-3p mimic plasmids had lower activity than other cells(P<0.05).The cell viability at 12,24,36 and 48 h was as follows:the miR-203a-3p inhibition group>the control group and the negative control group>the miR-203a-3p simulation group(all P<0.05).The cell mi-gration distance and the number of invasive cells were in the following order:the miR-203a-3p inhibition group>the con-trol group and negative control group>the miR-203a-3p simulation group(all P<0.05).Conclusion High expression of miR-203a-3p inhibits the migration and invasion of esophageal cancer cells,and the mechanism may be related to the tar-geted negative regulation of Survivin.

关键词

微小核糖核酸203a-3p/食管癌/Survivin/细胞侵袭/细胞迁移

Key words

miR-203a-3p/esophageal carcinoma/Survivin/cell invasion/cell migration

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基金项目

河北省医学科学研究重点课题计划项目(20210985)

出版年

2024
山东医药
山东卫生报刊社

山东医药

CSTPCD
影响因子:1.225
ISSN:1002-266X
参考文献量19
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