miR-449a对食管鳞癌细胞Eca-109增殖、迁移、侵袭的影响及机制

目的探讨miR-449a对食管鳞癌细胞Eca-109增殖、迁移、侵袭的影响及其可能的机制。方法将食管鳞癌细胞Eca-109分为miR-449a组和NC组,分别转染miR-449a-mimics和NC-mimics;采用实时荧光定量PCR法检测两组miR-449a相对表达量,miR-449a组miR-449a相对表达量明显高于NC组,证实上调miR-449a表达的Eca-109细胞模型建立成功。采用CCK-8法、平板克隆形成试验检测两组细胞增殖能力（分别以培养12、24、48 h吸光度值和克隆数表示）,细胞划痕试验检测细胞迁移能力（以细胞之间的距离表示）,Transwell小室试验检测细胞侵袭能...

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Bibliographic Details
Published in山东医药 Vol. 57; no. 16; pp. 24 - 27
Main Author 王强强 王梦洁 张玉虹 李皓 高飞 刘阳晨
Format Magazine Article
LanguageChinese
Published 蚌埠医学院研究生部,安徽蚌埠 233000 2017
泰兴市人民医院%泰兴市人民医院
Subjects
E2F3基因
微小RNA-449a
细胞侵袭
细胞增殖
细胞迁移
食管鳞癌
E2F3 gene
食管鳞癌
微小RNA-449a
细胞迁移
cell proliferation
microRNA-449a
cell migration
E2F3基因
esophageal squamous-cell carcinoma
cell invasion
细胞侵袭
细胞增殖
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ISSN1002-266X
DOI10.3969/j.issn.1002-266X.2017.16.007

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Summary:目的探讨miR-449a对食管鳞癌细胞Eca-109增殖、迁移、侵袭的影响及其可能的机制。方法将食管鳞癌细胞Eca-109分为miR-449a组和NC组,分别转染miR-449a-mimics和NC-mimics;采用实时荧光定量PCR法检测两组miR-449a相对表达量,miR-449a组miR-449a相对表达量明显高于NC组,证实上调miR-449a表达的Eca-109细胞模型建立成功。采用CCK-8法、平板克隆形成试验检测两组细胞增殖能力（分别以培养12、24、48 h吸光度值和克隆数表示）,细胞划痕试验检测细胞迁移能力（以细胞之间的距离表示）,Transwell小室试验检测细胞侵袭能力（以侵袭到下室的细胞数量表示）,流式细胞仪检测细胞周期比例。采用实时荧光定量PCR法和Western blotting法分别检测两组miR-449a直接靶基因E2F3 mRNA及蛋白相对表达量。结果 miR-449a组细胞培养12、24、48 h时的吸光度值及克隆数、细胞之间的距离、侵袭到下室的细胞数量均低于NC组（P〈0.05或〈0.01）。miR-449a组G1期细胞比例高于NC组,S期细胞比例低于NC组（P均〈0.05）;两组G2/M期细胞比例比较差异无统计学意义（P〉0.05）。miR-449a组E2F3 mRNA及蛋白相对表达量均低于NC组（P均〈0.01）。结论 miR-449a可抑制食管鳞癌细胞Eca-109的增殖、迁移和侵袭;抑制靶基因E2F3表达可能是其作用机制。
Bibliography:37-1156/R
esophageal squamous-cell carcinoma; microRNA-449a; E2F3 gene; cell proliferation; cell migration; cell invasion
WANG Qangqiang1, WANG Mengjie, ZHANG Yuhong, LI Hao, GAO Fei, LIU Yangchen （1 Bengbu Medical College, Bengbu 233000, China）
Objective To investigate the effects of miR-449a on cell proliferation, migration and invasion of esophageal squamous-cell carcinoma cell line Eca-109 and its mechanism.Methods Eca-109 cells were divided into the miR-449a group and the negative control （NC） group, in which miR-449a mimics and NC-mimics were transfected into Eca-109 cells, respectively.Real-time fluorescent quantitative PCR was performed to detect the relative expression of miR-449a in Eca-109 cells.When the expression of miR-449a in Eca-109 cells of the miR-449a group was higher than that of the NC group, indicating Eca-109 cell models with up-regulated miR-449a expression were successfully established.The cell proliferation abilities of the two groups were analyzed by CCK-8 and colony formation assay, w
ISSN:1002-266X
DOI:10.3969/j.issn.1002-266X.2017.16.007