miR-449a对食管鳞癌细胞Eca-109增殖、迁移、侵袭的影响及机制

发布时间：2018-05-11 02:13

本文选题：食管鳞癌 + 微小RNA-a　；参考：《山东医药》2017年16期

【摘要】：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(P0.05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(P0.05)。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。
[Abstract]:Objective to investigate the effect of miR-449a on the proliferation, migration and invasion of Eca-109 in esophageal squamous cell carcinoma cells and its possible mechanism. Methods the Eca-109 cells were divided into miR-449a and NC groups, miR-449a-mimics and NC-mimics were transfected respectively, and the relative expression of two groups of miR-449a and miR-449a relative expression in the miR-449a group were detected by real time fluorescence quantitative PCR method. The Eca-109 cell model for up regulation of miR-449a was successfully established. The Eca-109 cell proliferation ability of two groups was detected by CCK-8 method. The cell proliferation ability was detected by the culture of 12,24,48 h absorbency and clone number respectively. The cell migration energy was detected by the cell scratch test (expressed by the distance between cells), and the Transwell chamber test was used. The cell invasiveness of the cells (the number of cells invaded in the lower chamber) and the cell cycle ratio were detected by flow cytometry. The E2F3 mRNA and the relative expression of the two groups of miR-449a direct target genes were detected by real-time fluorescence quantitative PCR and Western blotting. The number of clones, distance between cells, and the number of cells that invaded the lower chamber were lower than that of group NC (P0.05 or 0.01), and the proportion of G1 cells in group.MiR-449a was higher than that in NC group, and the proportion of cells in S stage was lower than that in NC group (P 0.05), and there was no significant difference in the proportion of G2/M phase cells between the two groups (P0.05).MiR-449a group and the relative expression of protein was lower than that of the group (0.01 Conclusion miR-449a can inhibit the proliferation, migration and invasion of esophageal squamous cell carcinoma Eca-109, and inhibit the expression of target gene E2F3 may be its mechanism.

【作者单位】：蚌埠医学院研究生部;泰兴市人民医院;
【基金】：江苏省“333工程”科研项目(BRA2015225)
【分类号】：R735.1

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