Abstract

MicroRNAs epigenetically regulate physiological and pathological processes. Previously, we found that miR-204-5p is expressed at low levels in melanoma cells, and an increase in its level leads to a change in proliferation, migration, and invasion of these cancer cells. Now, using bioinformatics analysis, it has been shown that the target of miR-204-5p is FOXC1 transcription factor, which is implicated in carcinogenesis. Using the luciferase reporter assay, it was found that miR-204-5p suppresses expression of the FOXC1 gene by binding to its 3' non-coding region. Transfection of small interfering RNA (siRNA) targeting FOXC1 into melanoma cells caused a decrease in miR-204-5p levels, which is consistent with the generally accepted concept of feedback regulation of miRNA expression by target genes. According to the results of the MTT test and fluorescence microscopy, the proliferation level of melanoma cells under the influence of siRNA to FOXC1 decreased 72 h after transfection. Changes in the ratio of cells by cell cycle phase were analyzed using flow cytometry. Regulatory relationships between FOXC1 and miR-204-5p, and an inhibitory effect of FOXC1 knockdown on melanoma cell proliferation were revealed. Based on the results, it can be assumed that miR-204-5p regulates proliferation of melanoma cells by affecting FOXC1 expression.

中文翻译：

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FOXC1 介导的 miR-204-5p 对黑色素瘤细胞增殖的影响

摘要

MicroRNAs 表观遗传调节生理和病理过程。此前，我们发现miR-204-5p在黑色素瘤细胞中的表达水平较低，其水平的升高会导致这些癌细胞的增殖、迁移和侵袭发生变化。现在，使用生物信息学分析，已经表明 miR-204-5p 的靶标是 FOXC1 转录因子，它与致癌作用有关。使用荧光素酶报告基因检测，发现 miR-204-5p通过与其 3' 非编码区结合来抑制FOXC1基因的表达。转染靶向FOXC1的小干扰 RNA (siRNA)进入黑色素瘤细胞导致 miR-204-5p 水平降低，这与普遍接受的靶基因反馈调节 miRNA 表达的概念一致。根据MTT试验和荧光显微镜检查结果，转染后72h黑色素瘤细胞在针对FOXC1的siRNA影响下的增殖水平下降。使用流式细胞术分析细胞周期阶段细胞比例的变化。揭示了FOXC1和 miR-204-5p之间的调节关系，以及FOXC1敲低对黑色素瘤细胞增殖的抑制作用。根据结果​​，可以假设 miR-204-5p 通过影响FOXC1表达来调节黑色素瘤细胞的增殖。