PO.MCB10.01 · 分子与细胞生物学
褪黑素对乳腺癌细胞系中mir-655-3p的亚型依赖性调控
Subtype-dependent modulation of mir-655-3p by melatonin in breast cancer cell lines
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
背景：乳腺癌（BC）是一种高度异质性的疾病，由不同的分子亚型组成，这些亚型影响肿瘤行为和治疗反应。褪黑素（MEL）因其抑癌特性而被广泛研究，部分通过调控microRNA（miRNA）表达介导——miRNA是参与增殖、侵袭和转移的基因的关键转录后调控因子。其中，miR-655-3p被描述为一种潜在的抑癌miRNA，在多种癌症类型中与侵袭性降低相关。然而，MEL对不同BC亚型中miR-655-3p表达的影响仍不明确。因此，我们旨在评估MEL处理对具有不同分子特征的BC细胞系（包括luminal A和三阴性（TN）亚型）中miR-655-3p表达的影响。
方法：将人BC细胞系MCF-7和T47D（luminal A；ER+/PR+）以及MDA-MB-231（TN）在标准条件下培养，并用3000 μM MEL处理或不处理（对照组-CC）。使用mirVana™ miRNA分离试剂盒提取总RNA。使用High-Capacity cDNA逆转录试剂盒进行cDNA合成，并使用特异性TaqMan®试剂通过qRT-PCR定量miR-655-3p表达。使用ExpressionSuite™软件v1.3获得相对表达水平以及对内源性对照的归一化。
结果：与未处理对照相比，MEL处理导致MDA-MB-231（TN）细胞中miR-655-3p显著上调。相反，在相同MEL暴露条件下，luminal A细胞系（MCF-7和T47D）显示miR-655-3p表达显著下降。
结论：褪黑素在BC分子亚型间差异性调控miR-655-3p表达，在TN细胞中增加其表达，而在luminal A细胞中降低其表达。这些相反的表达模式提示，MEL对miR-655-3p的调控作用是亚型依赖性的，可能反映了与激素受体状态相关的信号通路差异。总之，这些发现表明miR-655-3p可作为对褪黑素亚型特异性反应的生物标志物，以及参与调控肿瘤侵袭性的潜在治疗靶点。有必要开展进一步研究以鉴定其直接基因靶点并阐明该调控的生物学意义。
查看英文原文 English abstract
Background: Breast cancer (BC) is a highly heterogeneous disease composed of distinct molecular subtypes that influence tumor behavior and therapeutic response. Melatonin (MEL) has been widely investigated for its oncostatic properties, partly mediated through the modulation of microRNA (miRNA) expression-key post-transcriptional regulators of genes involved in proliferation, invasion, and metastasis. Among these, miR-655-3p has been described as a potential tumor-suppressive miRNA associated with reduced aggressiveness in several cancer types. However, the effects of MEL on miR-655-3p expression across different BC subtypes remain poorly defined. Therefore, we aimed to evaluate the impact of MEL treatment on miR-655-3p expression in BC cell lines with distinct molecular profiles, including luminal A and triple-negative (TN) subtypes.
Methods: Human BC cell lines MCF-7 and T47D (luminal A; ER+/PR+) and MDA-MB-231 (TN) were cultured under standard conditions and treated with 3000 μM MEL or left untreated (control group - CC). Total RNA was extracted using the mirVana™ miRNA Isolation Kit. cDNA synthesis was performed with the High-Capacity cDNA Reverse Transcription Kit, and miR-655-3p expression was quantified by qRT-PCR using specific TaqMan® assays. Relative expression levels and normalization to endogenous controls were obtained using ExpressionSuite™ Software v1.3.
Results: MEL treatment resulted in a significant upregulation of miR-655-3p in MDA-MB-231 (TN) cells compared with untreated controls. In contrast, luminal A cell lines (MCF-7 and T47D) exhibited a significant decrease in miR-655-3p expression under the same MEL exposure conditions.
Conclusion: Melatonin differentially modulates miR-655-3p expression across BC molecular subtypes, increasing its expression in TN cells while reducing it in luminal A cells. These opposite expression patterns suggest that MEL's regulatory effects on miR-655-3p are subtype-dependent, potentially reflecting differences in signaling pathways associated with hormone receptor status. Together, these findings indicate that miR-655-3p may serve as a biomarker of subtype-specific responses to melatonin and a potential therapeutic target involved in the modulation of tumor aggressiveness. Further studies are warranted to identify its direct gene targets and elucidate the biological relevance of this regulation.
利益披露 Disclosure
K. C. Carvalho, None.