Shi Runlin, Xiao Haibing, Yang Tao, Chang Lei, Tian Yuanfeng, Wu Bolin, Xu Hua
Department of Urology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China.
Front Med. 2014 Dec;8(4):456-63. doi: 10.1007/s11684-014-0353-z. Epub 2014 Nov 3.
microRNAs (miRNAs) have played a key role in human tumorigenesis, tumor progression, and metastasis. On the one hand, miRNAs are aberrantly expressed in many types of human cancer; on the other hand, miRNAs can function as tumor suppressors or oncogenes that target many cancer-related genes. This study aimed to investigate the effects of miRNA-200c (miR-200c) on the biological behavior and mechanism of proliferation, migration, and invasion in the prostate cancer cell line Du145. In this study, Du145 cells were transfected with miR-200c mimics or negative control miR-NC by using an X-tremeGENE siRNA transfection reagent. The relative expression of miR-200c was measured by RT-PCR. The proliferation, migration, and invasion abilities of Du145 cells were detected by CCK8 assays, migration assays and invasion assays, respectively. The expressions of ZEB1, E-cadherin, and vimentin were observed by western blot. Results showed that DU145 cells exhibited a high expression of miR-200c compared with immortalized normal prostate epithelial cell RWPE-1. Du145 cells were then transfected with miR-200c mimics and displayed lower abilities of proliferation, migration, and invasion than those transfected with the negative control. The protein levels of ZEB1 and vimentin were expressed at a low extent in Du145 cells, which were transfected with miR-200c mimics; by contrast, E-cadherin was highly expressed. Hence, miR-200c could significantly inhibit the proliferation of the prostate cancer cell line Du145; likewise, miR-200c could inhibit migration and invasion by epithelial-mesenchymal transition.
微小RNA(miRNA)在人类肿瘤发生、肿瘤进展和转移过程中发挥了关键作用。一方面,miRNA在多种人类癌症中异常表达;另一方面,miRNA可作为靶向许多癌症相关基因的肿瘤抑制因子或癌基因发挥作用。本研究旨在探讨miRNA-200c(miR-200c)对前列腺癌细胞系Du145增殖、迁移和侵袭生物学行为及机制的影响。在本研究中,使用X-tremeGENE siRNA转染试剂将miR-200c模拟物或阴性对照miR-NC转染至Du145细胞。通过RT-PCR检测miR-200c的相对表达。分别通过CCK8检测、迁移检测和侵袭检测来检测Du145细胞的增殖、迁移和侵袭能力。通过蛋白质免疫印迹法观察ZEB1、E-钙黏蛋白和波形蛋白的表达。结果显示,与永生化正常前列腺上皮细胞RWPE-1相比,DU145细胞呈现出较高的miR-200c表达。随后将miR-200c模拟物转染至Du145细胞,其增殖、迁移和侵袭能力低于转染阴性对照的细胞。在转染了miR-200c模拟物的Du145细胞中,ZEB1和波形蛋白的蛋白水平低表达;相比之下,E-钙黏蛋白高表达。因此,miR-200c可显著抑制前列腺癌细胞系Du145的增殖;同样,miR-200c可通过上皮-间质转化抑制迁移和侵袭。