Department of Medical Genetics, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran.
Institute for Hygiene and Applied Immunology, Center for Pathophysiology, Infectiology, and Immunology, Medical University of Vienna, Molecular Immunology Unit, Vienna, Austria.
J Cell Biochem. 2018 Aug;119(8):6408-6417. doi: 10.1002/jcb.26577. Epub 2018 May 8.
In several cancers, microRNA (miRNAs) play vital roles in tumor initiation, drug resistance, and metastasis. The aim of this study was to examine the expression levels of miR-4301 in human breast cancer and investigate whether its potential roles involved targeting Dopamine receptor D2 (DRD2). Real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) was also used to examine the expression levels of miR-4301 in human breast cancer cell lines MDA-MB-231, MCF-7, and SKBR3. In these cell lines, MTT assay, immunofluorescence staining, caspase assay, proliferation assay, and flow cytometry were conducted to explore the potential functions of miR-4301. The effects of modulating miR-4301 on transcription levels of DRD2 were subsequently confirmed via qRT-PCR. miR-4301 expression levels were significantly decreased in human breast cancer specimens and cell lines (P < 0.05). Transfection of miR-4301 in breast cancer cells suppressed cell proliferation and induced apoptosis. Expression analysis indicated that miR-4301 was inversely correlated with DRD2 expression in breast cancer specimens. qRT-PCR showed that miR-4301 negatively regulated DRD2 expression. Downregulation of DRD2 expression in MDA-MB-231, MCF-7, and SKBR3 cells suppressed cell proliferation and promoted apoptosis.
在几种癌症中,microRNA(miRNA)在肿瘤起始、耐药和转移中发挥重要作用。本研究旨在检测人乳腺癌中 miR-4301 的表达水平,并探讨其潜在作用是否涉及靶向多巴胺受体 D2(DRD2)。实时定量逆转录聚合酶链反应(qRT-PCR)也用于检测人乳腺癌细胞系 MDA-MB-231、MCF-7 和 SKBR3 中 miR-4301 的表达水平。在这些细胞系中,通过 MTT 测定、免疫荧光染色、半胱天冬酶测定、增殖测定和流式细胞术来研究 miR-4301 的潜在功能。随后通过 qRT-PCR 证实了调节 miR-4301 对 DRD2 转录水平的影响。miR-4301 的表达水平在人乳腺癌标本和细胞系中明显降低(P<0.05)。转染 miR-4301 可抑制乳腺癌细胞的增殖并诱导细胞凋亡。表达分析表明,miR-4301 在乳腺癌标本中与 DRD2 表达呈负相关。qRT-PCR 显示 miR-4301 负调控 DRD2 表达。下调 MDA-MB-231、MCF-7 和 SKBR3 细胞中的 DRD2 表达可抑制细胞增殖并促进细胞凋亡。