Li Hong, Lian Bin, Liu Yaobang, Chai Dahai, Li Jinping
Department of Surgical Oncology, General Hospital of Ningxia Medical University, Yinchuan, Ningxia Hui Autonomous Region 750004, P.R. China.
Oncol Lett. 2020 Dec;20(6):277. doi: 10.3892/ol.2020.12140. Epub 2020 Sep 23.
Breast cancer (BC) is one of the most common malignant tumors among women worldwide. MicroRNAs (miRs) may be involved in several types of human cancer, including gastric, liver, lung and breast cancer. The aim of the present study was to investigate the effect of miR-1297 on MDA-MB-231 cell epithelial-mesenchymal transition (EMT) and proliferation, and the underlying molecular mechanisms. MDA-MB-231 cells were transfected with miR-1297 inhibitor or inhibitor control for 48 h. Subsequently, MTT and flow cytometry assays indicated that miR-1297 inhibitor significantly decreased cell proliferation and induced apoptosis compared with the inhibitor control group. In addition, reverse transcription-quantitative PCR and western blotting suggested that miR-1297 inhibitor suppressed EMT in MDA-MB-231 cells compared with the inhibitor control group. TargetScan bioinformatics analysis and a dual-luciferase reporter gene assay were performed, which predicted that miR-1297 directly targeted fatty acid 2-hydroxylase (FA2H). Furthermore, MDA-MB-231 cells were transfected with control-plasmid or FA2H-plasmid for 48 h. The results demonstrated that FA2H overexpression decreased MDA-MB-231 cell proliferation and increased apoptosis compared with the control-plasmid group. Additionally, FA2H-plasmid increased E-cadherin expression levels, and reduced N-cadherin and matrix metalloproteinase 9 expression levels at both the protein and mRNA level compared with control-plasmid. Finally, MDA-MB-231 cells were transfected with control-small interfering (si)RNA, FA2H-siRNA, inhibitor control, miR-1297 inhibitor, miR-1297 inhibitor + control siRNA or miR-1297 inhibitor + FA2H-siRNA, and the results suggested that the biological effects of miR-1297 inhibitor were reversed by co-transfection with FA2H siRNA. In conclusion, the present study indicated that miR-1297/FA2H might serve as a novel potential biomarker and therapeutic target for BC.
乳腺癌（BC）是全球女性中最常见的恶性肿瘤之一。微小RNA（miR）可能参与多种人类癌症，包括胃癌、肝癌、肺癌和乳腺癌。本研究的目的是探讨miR-1297对MDA-MB-231细胞上皮-间质转化（EMT）和增殖的影响及其潜在的分子机制。将miR-1297抑制剂或抑制剂对照转染MDA-MB-231细胞48小时。随后，MTT和流式细胞术分析表明，与抑制剂对照组相比，miR-1297抑制剂显著降低细胞增殖并诱导细胞凋亡。此外，逆转录-定量PCR和蛋白质印迹表明，与抑制剂对照组相比，miR-1297抑制剂抑制了MDA-MB-231细胞的EMT。进行了TargetScan生物信息学分析和双荧光素酶报告基因检测，预测miR-1297直接靶向脂肪酸2-羟化酶（FA2H）。此外，将对照质粒或FA2H质粒转染MDA-MB-231细胞48小时。结果表明，与对照质粒组相比，FA2H过表达降低了MDA-MB-231细胞增殖并增加了细胞凋亡。此外，与对照质粒相比，FA2H质粒在蛋白质和mRNA水平上均增加了E-钙黏蛋白的表达水平，并降低了N-钙黏蛋白和基质金属蛋白酶9的表达水平。最后，将对照小干扰（si）RNA、FA2H-siRNA、抑制剂对照、miR-1297抑制剂、miR-1297抑制剂+对照siRNA或miR-1297抑制剂+FA2H-siRNA转染MDA-MB-231细胞，结果表明，与FA2H siRNA共转染可逆转miR-1297抑制剂的生物学效应。总之，本研究表明，miR-1297/FA2H可能作为BC的一种新的潜在生物标志物和治疗靶点。
