Xia Wei, Zhou JueYu, Luo HaiBo, Liu YunZhou, Peng CanCan, Zheng WenLing, Ma WenLi
Institute of Genetic Engineering, Southern Medical University, Guangzhou, 510515 People's Republic of China.
Department of Clinical Laboratory, No.421 Hospital of PLA, Guangzhou, People's Republic of China.
Cancer Cell Int. 2017 Jan 25;17:14. doi: 10.1186/s12935-017-0383-0. eCollection 2017.
MicroRNAs are a class of small non-coding RNAs that are involved in many important physiological and pathological processes by regulating gene expression negatively. The purpose of this study was to investigate the effect of miR-32 on cell proliferation, migration and apoptosis and to determine the functional connection between miR-32 and FBXW7 in breast cancer.
In this study, quantitative RT-PCR was used to evaluate the expression levels of miR-32 in 27 breast cancer tissues, adjacent normal breast tissues and human breast cancer cell lines. The biological functions of miR-32 in MCF-7 breast cancer cells were determined by cell proliferation, apoptosis assays and wound-healing assays. In addition, the regulation of FBXW7 by miR-32 was assessed by qRT-PCR, Western blot and luciferase reporter assays.
MiR-32 was frequently overexpressed in breast cancer tissue samples and cell lines as was demonstrated by qRT-PCR. Moreover, the up-regulation of miR-32 suppressed apoptosis and promoted proliferation and migration, whereas down-regulation of miR-32 showed an opposite effect. Dual-luciferase reporter assays showed that miR-32 binds to the 3'-untranslated region of FBXW7, suggesting that FBXW7 is a direct target of miR-32. Western blot analysis showed that over-expression of miR-32 reduced FBXW7 protein level. Furthermore, an inverse correlation was found between the expressions of miR-32 and FBXW7 mRNA levels in breast cancer tissues. Knockdown of FBXW7 promoted proliferation and motility and suppressed apoptosis in MCF-7 cells.
Taken together, the present study suggests that miR-32 promotes proliferation and motility and suppresses apoptosis of breast cancer cells through targeting FBXW7.
微小RNA是一类小的非编码RNA,通过负向调节基因表达参与许多重要的生理和病理过程。本研究旨在探讨miR-32对细胞增殖、迁移和凋亡的影响,并确定miR-32与乳腺癌中FBXW7之间的功能联系。
在本研究中,采用定量逆转录聚合酶链反应(qRT-PCR)评估27例乳腺癌组织、癌旁正常乳腺组织及人乳腺癌细胞系中miR-32的表达水平。通过细胞增殖、凋亡检测和伤口愈合实验确定miR-32在MCF-7乳腺癌细胞中的生物学功能。此外,通过qRT-PCR、蛋白质免疫印迹法(Western blot)和荧光素酶报告基因检测评估miR-32对FBXW7的调控作用。
qRT-PCR结果显示,miR-32在乳腺癌组织样本和细胞系中经常过表达。此外,miR-32的上调抑制细胞凋亡,促进增殖和迁移,而miR-32的下调则显示出相反的效果。双荧光素酶报告基因检测表明,miR-32与FBXW7的3'-非翻译区结合,提示FBXW7是miR-32的直接靶点。Western blot分析显示,miR-32的过表达降低了FBXW7蛋白水平。此外,在乳腺癌组织中发现miR-32与FBXW7 mRNA水平的表达呈负相关。敲低FBXW7可促进MCF-7细胞的增殖和运动能力,并抑制其凋亡。
综上所述,本研究表明miR-32通过靶向FBXW7促进乳腺癌细胞的增殖和运动能力,并抑制其凋亡。