[1]雷周满,菅书明,冯东升.微RNA-139-5p对胃癌细胞增殖和侵袭的影响[J].新乡医学院学报,2021,38(1):026-30.[doi:10.7683/xxyxyxb.2021.01.005]
　LEI Zhouman,JIAN Shuming,FENG Dongsheng.Effect of microRNA-139-5p on the proliferation and invasion of gastric cancer cells[J].Journal of Xinxiang Medical University,2021,38(1):026-30.[doi:10.7683/xxyxyxb.2021.01.005]
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微RNA-139-5p对胃癌细胞增殖和侵袭的影响
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《新乡医学院学报》[ISSN:1004-7239/CN:41-1186/R]

卷:
38
期数:
2021年1
页码:
026-30
栏目:
基础研究
出版日期:
2021-01-05

文章信息/Info

Title:
Effect of microRNA-139-5p on the proliferation and invasion of gastric cancer cells
作者:
雷周满; 菅书明; 冯东升
（许昌市中心医院普外科，河南　许昌　461000）
Author(s):
LEI Zhouman; JIAN Shuming; FENG Dongsheng
(Department of General Surgery,Xuchang Central Hospital,Xuchang 461000,Henan Province,China)
关键词:
miR-139-5p; 胃癌; 细胞增殖; 细胞侵袭; Wnt/β-catenin信号通路
Keywords:
miR-139-5pgastric carcinomacell proliferationcell invasionWnt/β-catenin signaling pathway
分类号:
R735.2
DOI:
10.7683/xxyxyxb.2021.01.005
文献标志码:
A
摘要:
目的　探讨微RNA（miR）-139-5p对胃癌细胞增殖和侵袭的影响及可能的作用机制。方法　采用实时荧光定量聚合酶链反应（PCR）检测永生化胃黏膜上皮细胞GES1和胃癌细胞SGC-7901、AGS和BGC-823中miR-139-5p的表达，Western blot法检测GES1、SGC-7901、AGS和BGC-823细胞中Wnt3a、β-catenin和细胞周期蛋白D1（cyclin D1）蛋白表达。收集对数生长期胃癌细胞SGC-7901，待细胞贴壁生长融合度达50%～60%时将细胞分为miR-139-5p mimics组、miR-NC组和空白对照组，利用脂质体Lipofectamine 2000分别将miR-139-5p mimics、阴性对照质粒转染至miR-139-5pmimics组和miR-NC组SGC-7901细胞，空白对照组细胞不转染任何质粒。采用实时荧光定量PCR法检测3组SGC-7901细胞中miR-139-5p的表达，甲基噻唑基四唑法检测3组SGC-7901细胞活力，Transwell实验检测3组SGC-7901细胞侵袭能力，Western blot法检测3组SGC-7901细胞中Wnt3a、β-catenin和cyclin D1蛋白表达。结果　SGC-7901、AGS、BGC-823和GES1细胞中miR-139-5p相对表达量分别为0.15±0.02、0.62±0.06、0.50±0.05、1.24±0.14；SGC-7901、AGS和BGC-823细胞中miR-139-5p相对表达量显著低于GES1细胞（P<0.05），sgc-7901细胞中mir-139-5p相对表达量显著低于ags、bgc-823细胞（>P<0.05），ags与bgc-823细胞中mir-139-5p相对表达量比较差异无统计学意义（>P>0.05）。空白对照组、miR-NC组和miR-139-5p mimics组SGC-7901细胞中miR-139-5p相对表达量分别为1.05±0.10、0.96±0.07、2.01±0.23；miR-139-5p mimics组SGC-7901细胞中miR-139-5p相对表达量显著高于空白对照组和miR-NC组（P<0.05），空白对照组与mir-nc组细胞中mir-139-5p相对表达量比较差异无统计学意义（>P＞0.05）。转染后72、96 h，miR-139-5p mimics组SGC-7901细胞活力显著低于空白对照组和miR-NC组（P<0.05），空白对照组与mir-nc组sgc-7901细胞活力比较差异无统计学意义（>P>0.05）。空白对照组、miR-NC组和miR-139-5p mimics组穿膜细胞数分别为98.67±10.26、101.45±8.24、49.33±5.12；miR-139-5p mimics组穿膜细胞数显著少于空白对照组和miR-NC组（P<0.05），空白对照组与mir-nc组穿膜细胞数比较差异无统计学意义（>P＞0.05）。miR-139-5p mimics组SGC-7901细胞中Wnt3a、β-catenin和cyclin D1蛋白相对表达量显著低于空白对照组和miR-NC组（P<0.05），空白对照组与mir-nc组sgc-7901细胞中wnt3a、β-catenin和cyclin d1蛋白相对表达量比较差异无统计学意义（P＞0.05）。结论　胃癌细胞中miR-139-5p表达下调，miR-139-5p可能通过阻断Wnt/β-catenin信号通路传导而抑制胃癌细胞SGC-7901的增殖和侵袭，miR-139-5p有望成为治疗胃癌的新靶点。undefinedundefinedundefinedundefinedundefinedundefined/html>
Abstract:
Objective　To investigate the effect of microRNA（miR）-139-5p on the proliferation and invasion of gastric cancer cells and its possible mechanism.Methods　　The expression of miR-139-5p in immortalized gastric epithelial cells GES1 and gastric cancer cells SGC-7901,AGS and BGC-823 was detected by real time fluorescent quantitative polymerase chain reaction (PCR).The expression of Wnt3a,β-catenin and cyclin D1 protein in GES1,SGC-7901,AGS and BGC-823 cells was detected by Western blot method.The SGC-7901 cells in logarithmic growth phase were collected and divided into miR-139-5p mimics group,miR-NC group and blank control group when the confluence of cell adhesion growth reached 50%-60%.The miR-139-5p mimics and negative control plasmid were transfected into SGC-7901 cells in the miR-139-5p mimics group and miR-NC group by Lipofectamine 2000,respectively.No plasmid was transfected into SGC-7901 cells in the blank control group.The expression of miR-139-5p in SGC-7901 cells was detected by real time fluorescent quantitative PCR,the viability of SGC-7901 cells was detected by methyl thiazolyl tetrazolium assay,the invasion ability of SGC-7901 cells was detected by Transwell experiment,and the expressions of Wnt3a,β-catenin and cyclin D1 protein in SGC-7901 cells of the three groups were detected by Western blot method.Results　The relative expressions of miR-139-5p in SGC-7901,AGS,BGC-823 and GES1 cells was 0.15 ± 0.02,0.62±0.06,0.50 ± 0.05 and 1.24 ± 0.14,respectively.The relative expression of miR-139-5p in SGC-7901,AGS and BGC-823 cells was significantly lower than that in GES1 cells (P<0.05).The relative expression of miR-139-5p in SGC-7901 cells was significantly lower than that in AGS and BGC-823 cells (P<0.05).There was no significant difference in the relative expression of miR-139-5p between AGS and BGC-823 cells (P>0.05).The relative expression of miR-139-5p in SGC-7901 cells in the blank control group,miR-NC group and miR-139-5p mimics group was 1.05±0.10,0.96 ± 0.07 and 2.01 ± 0.23,respectively.The relative expression of miR-139-5p in SGC-7901 cells in the miR-139-5p mimics group was significantly higher than that in the blank control group and miR-NC group (P<0.05).There was no significant difference in the relative expression of miR-139-5p between the blank control group and miR-NC group(P>0.05).At 72 and 96 hours after transfection,the viability of SGC-7901 cells in the miR-139-5p mimics group was significantly lower than that in the blank control group and miR-NC group (P<0.05),but there was no significant difference in the viability of SGC-7901 cells between the blank control group and miR-NC group (P<0.05).The number of transmembrane cells in the blank control group,miR-NC group and miR-139-5p mimics group was 98.67 ± 10.26,101.45 ± 8.24 and 49.33 ± 5.12,respectively.The number of transmembrane cells in the miR-139-5p mimics group was significantly less than that in the blank control group and miR-NC group (P<0.05),but there was no significant difference in the number of transmembrane cells between the blank control group and miR-NC group (P>0.05).The relative expressions of Wnt3a,β-catenin and cyclin D1 protein in SGC-7901 cells in the miR-139-5p mimics group were significantly lower than those in the blank control group and miR-NC group (P<0.05).There was no significant difference in the relative expressions of Wnt3a,β-catenin and cyclin D1 protein between the blank control group and miR-NC group (P>0.05).Conclusion　The expression of miR-139-5p in gastric cancer cells is down-regulated,miR-139-5p may inhibit the proliferation and invasion of SGC-7901 cells by blocking Wnt/β-catenin signaling pathway,and miR-139-5p is expected to become a new target for the treatment of gastric cancer.

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更新日期/Last Update: 2021-01-05