Yuan Jianfen, Xiao Chunhong, Lu Huijun, Yu HaizHong, Hong Hong, Guo Chunyan, Wu Zhimei
Department of Clinical Laboratory, Nantong Traditional Chinese Medicine Hospital, Nantong, Jiangsu 226001, P.R. China.
Department of Clinical Laboratory, Nantong Tumor Hospital, Nantong, Jiangsu 226361, P.R. China.
Exp Ther Med. 2020 Apr;19(4):2741-2750. doi: 10.3892/etm.2020.8516. Epub 2020 Feb 11.
Radixin is an important member of the Ezrin-Radixin-Moesin protein family that is involved in cell invasion, metastasis and movement. microRNA (miR)-200b is a well-studied microRNA associated with the development of multiple tumors. Previous bioinformatics analysis has demonstrated that miR-200b has a complementary binding site in the 3'-untranslated region of radixin mRNA. The present study aimed to investigate the role of miR-200b in regulating radixin expression, cell proliferation and invasion in breast cancer. Breast cancer tissues at different Tumor-Node-Metastasis (TNM) stages were collected; breast tissues from patients with hyperplasia were used as a control. miR-200b and radixin mRNA expression levels were tested by reverse transcription-quantitative PCR. Radixin protein expression was detected by western blotting. The highly metastatic MDA-MB-231 cells were divided into four groups and transfected with a miR-negative control (NC), miR-200b mimic, small interfering (si)RNA-NC or siRNA targeting radixin. Cell invasion was evaluated by Transwell assay and cell proliferation was assessed by 5-ethynyl-2'-deoxyuridine staining. Compared with the control group, radixin mRNA expression was significantly higher in breast cancer tissues and increased with TNM stage. miR-200b expression levels exhibited the opposite trend. Radixin mRNA expression in breast cancer cells was notably higher, whereas miR-200b expression was lower compared with that in normal breast epithelial MCF-10A cells. The expression of radixin was higher, whereas miR-200b was lower in MDA-MB-231 cells compared with that in MCF-7 cells. miR-200b mimic or siRNA-radixin transfection downregulated the expression of radixin in MDA-MB-231 cells and attenuated the invasive and proliferative abilities of these cells. miR-200b-knockdown and radixin overexpression were associated with enhanced cell invasion in breast cancer. In conclusion, miR-200b regulates breast cancer cell proliferation and invasion by targeting radixin expression.
根蛋白是埃兹蛋白-根蛋白-膜突蛋白家族的重要成员,参与细胞侵袭、转移和运动。微小RNA(miR)-200b是一种研究充分的与多种肿瘤发生发展相关的微小RNA。先前的生物信息学分析表明,miR-200b在根蛋白mRNA的3'-非翻译区有一个互补结合位点。本研究旨在探讨miR-200b在调节乳腺癌中根蛋白表达、细胞增殖和侵袭方面的作用。收集不同肿瘤-淋巴结-转移(TNM)分期的乳腺癌组织;将增生患者的乳腺组织用作对照。通过逆转录定量PCR检测miR-200b和根蛋白mRNA表达水平。通过蛋白质印迹法检测根蛋白的蛋白表达。将高转移性的MDA-MB-231细胞分为四组,分别用miR阴性对照(NC)、miR-200b模拟物、小干扰(si)RNA-NC或靶向根蛋白的siRNA进行转染。通过Transwell实验评估细胞侵袭能力,通过5-乙炔基-2'-脱氧尿苷染色评估细胞增殖能力。与对照组相比,根蛋白mRNA在乳腺癌组织中的表达显著更高,且随TNM分期增加。miR-200b表达水平呈现相反趋势。与正常乳腺上皮MCF-10A细胞相比,乳腺癌细胞中根蛋白mRNA表达明显更高,而miR-200b表达更低。与MCF-7细胞相比,MDA-MB-231细胞中根蛋白表达更高,而miR-200b更低。miR-200b模拟物或siRNA-根蛋白转染下调了MDA-MB-231细胞中根蛋白的表达,并减弱了这些细胞的侵袭和增殖能力。miR-200b敲低和根蛋白过表达与乳腺癌细胞侵袭增强有关。总之,miR-200b通过靶向根蛋白表达来调节乳腺癌细胞的增殖和侵袭。