中文摘要：

目的 · 研究 hsa-miRNA124-3p 调控肺癌细胞 NCI-H460 增殖和迁移的作用及机制。方法 · 取临床肺癌及其癌旁组织标本 4 对， 分别检测组织中hsa-miRNA124-3p 及 Krüppel 样因子4（KLF4）蛋白含量。生物信息学预测hsa-miRNA124-3p 与 KLF4 基因3’-UTR 有理论结合位点，并通过荧光素酶报告基因实验进行验证。转染pshRNA-Sponge-miRNA124 或 pshRNA-KLF4 到 NCI-H460 细胞，转 染后48 h 确定细胞转染效率，MTT 检测 NCI-H460 细胞对数期增殖活性，划线法检测细胞迁移变化。结果 · 在检测的4 对标本中， hsa-miRNA124-3p 在肿瘤组织中的含量明显高于癌旁组织（P〈0.01），而肺癌组织中KLF4 蛋白表达明显低于癌旁组织（P〈0.01）。 生物信息学分析结果显示，hsa-miRNA124-3p 在 KLF4 基因3’-UTR 有 8 个碱基的理论结合位点“5’-UGCCUUAA-3’”。荧光素酶活 性检测数据显示，hsa-miRNA124-3p 可以结合于KLF4 基因3’-UTR 并对蛋白表达进行负调控。细胞转染后72 h，pshRNA-SpongemiRNA124 转染组细胞增殖活性明显受到抑制（P〈0.01）， pshRNA-KLF4 转染组细胞增殖活性较对照组增强（P〈0.05）， pshRNASponge-miRNA124 与 pshRNA-KLF4 共转染组细胞增殖活性与对照组比较，无显著差异（P〉0.05）。细胞迁移检测数据表明，不同处 理组细胞转染后72 h 细胞迁移能力变化与增殖活性的变化趋势完全一致。结论 · Hsa-miRNA124-3p 通过抑制KLF4 基因表达，增强 NCI-H460 细胞的增殖和迁移；敲减细胞内 hsa-miRNA124-3p，可以通过上调 KLF4 蛋白表达抑制 NCI-H460 细胞增殖和迁移活性。

英文摘要：

Objective · To study the regulation of hsa-miRNA124-3p on the proliferation and migration of human lung cancer NCI-H460 cells and its mechanism. Methods · Four pairs of lung cancer and para-carcinoma tissues were harvested in clinical and measured for hsa-miRNA124-3p and Krüppellike factor 4 （KLF4） levels. The theoretical binding site of hsa-miRNA124-3p in 3’-UTR of KLF4 was predicted by bioinformatics, and validated by luciferase report assay. NCI-H460 cells were transfected with pshRNA-Sponge-miRNA124 or pshRNA-KLF4, and 48 hours later, the proliferation of NCI-H460 cells after genetic intervention was assayed by the MTT method, and cell migration ability was observed by streak method. Results · For all four pairs of samples tested, hsa-miRNA124-3p was higher in the cancer tissues than in the adjacent tissue （P〈0.01）, and KLF4 protein was lower in the cancer tissues than in the adjacent tissue （P〈0.01）. The bioinformatic analysis showed there is a theoretical binding site （5’-UGCCUUAA-3’） of hsa-miRNA124-3p in 3’-UTR of KLF4. Luciferase activity assay showed that hsa-miRNA124-3p could bind to the 3’-UTR region of KLF4 gene and negatively regulate the expression of protein. The proliferation of NC-H460 cells was suppressed by transfection with pshRNA-Sponge-miRNA124 72 h after transfection （P〈0.05 ）. Compared with the control group, the proliferation activity of pshRNA-KLF4 transfection group was further enhanced （P〈0.05） There was no significant difference in the proliferation of pshRNA-Sponge-miRNA124 and pshRNA-KLF4 cotransfection group and the control group （P〉0.05）. The data of cell migration assay showed that the changes of cell migration ability were the same as proliferation activity of the cells in groups 72 h after transfection. Conclusion · Hsa-miRNA124-3p increases the proliferation and migration in NCI-H460 cells via suppressing the expression of KLF4, and reducing the content of miRNA124-3p in NC-H460 cells can inhibit cell proliferation and migr