Laboratory of Molecular Research in Cancer - LIMC, Faculdade de Medicina de Sao Jose do Rio Preto - FAMERP, Sao Jose do Rio Preto, SP, Brazil.
Graduate Program in Health Science, Faculdade de Medicina de Sao Jose do Rio Preto - FAMERP, Sao Jose do Rio Preto, SP, Brazil; Laboratory of Molecular Research in Cancer - LIMC, Faculdade de Medicina de Sao Jose do Rio Preto - FAMERP, Sao Jose do Rio Preto, SP, Brazil.
Life Sci. 2018 Sep 1;208:131-138. doi: 10.1016/j.lfs.2018.07.012. Epub 2018 Jul 7.
Breast cancer represents the second most prevalent tumor-related cause of death among women. Although studies have already been published regarding the association between breast tumors and miRNAs, this field remains unclear. MicroRNAs (miRNAs) are defined as non-coding RNA molecules, and are known to be involved in cell pathways through the regulation of gene expression. Melatonin can regulate miRNAs and genes related with angiogenesis. This hormone is produced naturally by the pineal gland and presents several antitumor effects. The aim of this study was to understand the action of melatonin in the regulation of miRNA-152-3p in vivo and in vitro.
In order to standardize the melatonin treatment in the MDA-MB-468 cells, we carried out the cell viability assay at different concentrations. PCR Array plates were used to identify the differentiated expression of miRNAs after the treatment with melatonin. The relative quantification of the target gene expression (IGF-IR, HIF-1α and VEGF) was performed by real-time PCR. For the tumor development, MDA-MB-468 cells were implanted in female BALB/c mice, and treated or not treated with melatonin. Moreover, the quantification of the target genes protein expression was performed by immunocytochemistry and immunohistochemistry.
Relative quantification shows that the melatonin treatment increases the gene expression of miR-152-3p and the target genes, and decreased protein levels of the genes both in vitro and in vivo.
Our results confirm the action of melatonin on the miR-152-3p regulation known to be involved in the progression of breast cancer.
乳腺癌是女性肿瘤相关死亡的第二大常见原因。尽管已经有研究报道了乳腺癌肿瘤与 microRNAs（miRNAs）之间的关联，但该领域仍不清楚。miRNAs 被定义为非编码 RNA 分子，已知通过调节基因表达参与细胞途径。褪黑素可以调节 miRNA 和与血管生成相关的基因。这种激素是由松果腺自然产生的，具有多种抗肿瘤作用。本研究旨在了解褪黑素在体内和体外调节 miRNA-152-3p 的作用。
为了规范 MDA-MB-468 细胞中褪黑素的治疗，我们在不同浓度下进行了细胞活力测定。使用 PCR 阵列板来鉴定褪黑素处理后 miRNA 的差异表达。通过实时 PCR 对靶基因表达（IGF-IR、HIF-1α 和 VEGF）的相对定量进行分析。为了进行肿瘤发展，将 MDA-MB-468 细胞植入雌性 BALB/c 小鼠体内，并对其进行或不进行褪黑素处理。此外，通过免疫细胞化学和免疫组织化学来检测靶基因蛋白表达的定量。
相对定量显示，褪黑素处理增加了 miR-152-3p 和靶基因的基因表达，并降低了体内外靶基因的蛋白水平。
我们的结果证实了褪黑素对 miR-152-3p 调节的作用，已知该调节参与乳腺癌的进展。
