Overexpression of miR-340-5p inhibits skin fibroblast proliferation by targeting Kruppel-like factor 2.
Overexpression of miR-340-5p inhibits skin fibroblast proliferation by targeting Kruppel-like factor 2.
复制标题
DOI：
10.2174/1389201020666190725112304
复制
发表时间：
2019
影响因子：
2.8
通讯作者：
Ling Chen;Qian Li;Xun Lu;Xiaohua Dong;Jingyun Li
中科院分区：
文献类型：
--
作者：
Ling Chen;Qian Li;Xun Lu;Xiaohua Dong;Jingyun Li
MicroRNA (miR)-340-5p已被确定在几种癌症中发挥关键作用。然而，miR-340-5p在皮肤成纤维细胞中的功能在很大程度上仍然未知。方法用携带22bp miR-340-5p的慢病毒感染正常皮肤成纤维细胞获得功能。CCK-8法检测细胞增殖。为了揭示其机制，我们使用了mRNA-seq。通过基因本体和KEGG通路进一步分析差异表达mrna。western blot检测蛋白水平。采用双荧光素酶报告基因法检测miR-340-5p与Kruppel样因子2 (KLF2) 3'UTR的直接结合。结果MiR-340-5p慢病毒感染可抑制正常皮肤成纤维细胞增殖。mRNA-seq数据显示，与对照组相比，miR-340-5p过表达组有41个mrna差异表达，其中22个转录本上调，19个转录本下调。基因本体和通路分析表明，miR-340-5p过表达与大分子生物合成过程、细胞大分子生物合成过程、膜和MAPK信号通路相关。生物信息学分析和荧光素酶报告基因分析表明，miR-340-5p与KLF2的3'UTR结合。miR-340-5p的强制表达降低了正常皮肤成纤维细胞中KLF2的表达。miR-340-5p过表达组KLF2可恢复皮肤成纤维细胞增殖。结论本研究表明miR-340-5p可能通过靶向KLF2抑制皮肤成纤维细胞增殖。这些发现有助于理解miR-340-5p在皮肤成纤维细胞中的功能。miR-340-5p可能成为预防瘢痕形成的治疗靶点。
OBJECTIVE MicroRNA (miR)-340-5p has been identified to play a key role in several cancers. However, the function of miR-340-5p in skin fibroblasts remains largely unknown. METHOD Gain of function was used by infecting the normal skin fibroblast cells with lentivirus carrying a 22 bp miR-340-5p. Cell proliferation was detected by CCK-8 assay. To uncover the mechanisms, mRNA-seq was used. Differentially expressed mRNAs were further analysed by gene ontology and KEGG pathway. The protein levels were analysed by western blot. Dual luciferase reporter assay was used to detect the direct binding of miR-340-5p with the 3'UTR of Kruppel like factor 2 (KLF2). RESULTS MiR-340-5p lentivirus infection suppressed normal skin fibroblast proliferation. The mRNA-seq data revealed that 41 mRNAs were differentially expressed, including 22 up-regulated and 19 down-regulated transcripts in the miR-340-5p overexpression group compared with control group. Gene ontology and pathway analysis revealed that miR-340-5p overexpression correlated with macromolecule biosynthetic process, cellular macromolecule biosynthetic process, membrane, and MAPK signaling pathway. Bioinformatics analysis and luciferase reporter assays showed that miR-340-5p binds to the 3'UTR of KLF2. Enforced expression of miR-340-5p decreased the expression of KLF2 in normal skin fibroblasts. Overexpression of KLF2 restores skin fibroblast proliferation in miR-340-5p overexpression group. CONCLUSION This study demonstrates that miR-340-5p may suppress skin fibroblast proliferation, and possibly through targeting KLF2. These findings could help in the understanding the function of miR-340-5p in skin fibroblast. And miR-340-5p could be a therapy target for preventing scarring.