Effects of miR-200c on the migration and invasion abilities of human prostate cancer Du145 cells and the corresponding mechanism

doi:10.1007/s11684-014-0353-z

Front. Med.

2014, Vol. 8

Issue (4) : 456-463 https://doi.org/10.1007/s11684-014-0353-z

RESEARCH ARTICLE

Effects of miR-200c on the migration and invasion abilities of human prostate cancer Du145 cells and the corresponding mechanism

Runlin Shi,Haibing Xiao,Tao Yang,Lei Chang,Yuanfeng Tian,Bolin Wu,Hua Xu(

)

Department of Urology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China

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Abstract

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.

Keywords miR-200c proliferation migration invasion prostate cancer Du145 cell ZEB1

Corresponding Author(s): Hua Xu

Issue Date: 18 December 2014

Cite this article:

Runlin Shi,Haibing Xiao,Tao Yang, et al. Effects of miR-200c on the migration and invasion abilities of human prostate cancer Du145 cells and the corresponding mechanism[J]. Front. Med., 2014, 8(4): 456-463.

URL:

https://academic.hep.com.cn/fmd/EN/10.1007/s11684-014-0353-z
https://academic.hep.com.cn/fmd/EN/Y2014/V8/I4/456

Fig.1 Expression of Du145 cells compared with that of human non-transformed prostate epithelial cell line RWPE-1; miR-200c expression in prostate cancer Du145 cells was reduced significantly. **P < 0.01.

Fig.2 Expressions of miR-200c and ZEB1 in the cells detected by qPCR. A reaction system (20 μl) and other systems were amplified and detected under the following conditions: 40 thermo cycles at 95 °C for 15 s and 60 °C for 20 s; these reactions were extended at 70 °C for 20 s. miR-200c may regulate ZEB1 expression on a transcriptional level, *P < 0.05 and ** P < 0.01. Difference was statistically significant.

Fig.3 CCK-8 assay revealing the proliferation of the prostate cancer cell line Du145. miR-200c could significantly suppress the proliferation of prostate cancer Du145 cells (P < 0.05; Fig. 3A). In vitro invasion assays showed that miR-200c could significantly suppress the invasion ability of prostate cancer cells Du145 (51.279 ± 11.338, * P < 0.05; Fig. 3B). In Transwell migration assay, cells were placed in a well at a density of 2×10⁵ cells/well; Du145 cells were stably transfected with miR-200c mimics and miR-220-NC. The cells then penetrated the lower surface of the filter without or with Matrigel. Hence, prostate cancer Du145 cells were stably transfected with miR-200c mimics (34.345 ± 6.238, * P < 0.05) presented an impaired migration ability compared with those transfected with miR-NC group (the negative control group; Fig. 3C).

Fig.4 Epithelial-to-mesenchymal transition. Western blot result indicated that the expressions of ZEB1 and vimentin were reduced when miR-200c was transfected in the cells; on the contrary, the expression of E-cadherin was increased. Difference was statistically significant.

Fig.5 Prediction of target genes showing that locus 5 of the 3′ UTR in ZEB1 could be combined with miR-200c.

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