Suppression of α‑methylacyl‑coenzyme A racemase by miR200c inhibits prostate adenocarcinoma cell proliferation and migration

Xie,Hanbing; Nie,Ling; Zhang,Mengni; Su,Zhengzheng; Chen,Xueqin; Xu,Miao; Gong,Jing; Chen,Ni; Zhou,Qiao

doi:10.3892/etm.2019.8406

Suppression of α‑methylacyl‑coenzyme A racemase by miR200c inhibits prostate adenocarcinoma cell proliferation and migration

Authors:
- Hanbing Xie
- Ling Nie
- Mengni Zhang
- Zhengzheng Su
- Xueqin Chen
- Miao Xu
- Jing Gong
- Ni Chen
- Qiao Zhou
View Affiliations

Affiliations: Pathology Department and State Key Laboratory of Biotherapy, West China Hospital, West China Medical School, Sichuan University, Chengdu, Sichuan 610041, P.R. China
Published online on: December 31, 2019 https://doi.org/10.3892/etm.2019.8406
Pages: 1806-1816
Copyright: © Xie et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Overexpression of α‑methylacyl‑coenzyme A racemase (AMACR/P504S) is a major abnormality that has been observed in prostate cancer, whereas microRNA (miRNA/miR) 200c, is downregulated. The aim of the present study was to explore whether miR200c was able to exert any regulatory effects on AMACR. To meet this aim, bioinformatics analysis was performed to identify potential binding sites for miR200c in the 3'‑untranslated region (3'‑UTR) of AMACR. Recombinant adenoviral and dual reporter gene assays were designed to examine the binding of miR200c to the potential seed sequences in the AMACR 3'‑UTR. Conventional reverse transcription (RT)‑PCR, RT‑quantitative (q)PCR and western blotting were also used to examine the regulatory effects of miR200c on AMACR at the mRNA and protein levels. Furthermore, Cell Counting Kit‑8, wound healing and Transwell assays were performed to investigate the biological effects of miR200c‑AMACR deregulation on prostate cancer cell proliferation, migration and invasion. It was revealed that miR200c post‑transcriptionally suppressed AMACR expression by interacting with the 90‑97 nucleotide sequence of the AMACR mRNA 3'‑UTR. Artificial overexpression of miR200c significantly downregulated the mRNA and protein levels of AMACR in DU145 and PC‑3 prostate cancer cells. Knockdown of AMACR by RNA interference, or overexpression of miR200c by recombinant adenoviral Ad‑miR200c, inhibited prostate cancer cell proliferation, migration and invasiveness. Taken together, the results of the present study revealed that miR200c may suppress the AMACR expression level post‑transcriptionally. The results also indicate that perturbation of the miR200c‑AMACR regulatory mechanism may be involved in prostate carcinogenesis and that this may be exploited in future therapeutic approaches to prostate cancer.

View Figures

View References

1	Knezevic J, Pfefferle AD, Petrovic I, Greene SB, Perou CM and Rosen JM: Expression of miR-200c in claudin-low breast cancer alters stem cell functionality, enhances chemosensitivity and reduces metastatic potential. Oncogene. 34:5997–6006. 2015. View Article : Google Scholar : PubMed/NCBI
2	Jurmeister S, Baumann M, Balwierz A, Keklikoglou I, Ward A, Uhlmann S, Zhang JD, Wiemann S and Sahin Ö: MicroRNA-200c represses migration and invasion of breast cancer cells by targeting actin-regulatory proteins FHOD1 and PPM1F. Mol Cell Biol. 32:633–651. 2012. View Article : Google Scholar : PubMed/NCBI
3	Zhou X, Wang Y, Shan B, Han J, Zhu H, Lv Y, Fan X, Sang M, Liu XD and Liu W: The downregulation of miR-200c/141 promotes ZEB1/2 expression and gastric cancer progression. Med Oncol. 32:4282015. View Article : Google Scholar : PubMed/NCBI
4	Valladares-Ayerbes M, Reboredo M, Medina-Villaamil V, Iglesias-Díaz P, Lorenzo-Patiño MJ, Haz M, Santamarina I, Blanco M, Fernández-Tajes J, Quindós M, et al: Circulating miR-200c as a diagnostic and prognostic biomarker for gastric cancer. J Transl Med. 10:1862012. View Article : Google Scholar : PubMed/NCBI
5	Toiyama Y, Hur K, Tanaka K, Inoue Y, Kusunoki M, Boland CR and Goel A: Serum miR-200c is a novel prognostic and metastasis-predictive biomarker in patients with colorectal cancer. Ann Surg. 259:735–743. 2014. View Article : Google Scholar : PubMed/NCBI
6	Gao YC and Wu J: MicroRNA-200c and microRNA-141 as potential diagnostic and prognostic biomarkers for ovarian cancer. Tumour Biol. 36:4843–4850. 2015. View Article : Google Scholar : PubMed/NCBI
7	Butz H, Szabó PM, Khella HW, Nofech-Mozes R, Patocs A and Yousef GM: miRNA-target network reveals miR-124as a key miRNA contributing to clear cell renal cell carcinoma aggressive behaviour by targeting CAV1 and FLOT1. Oncotarget. 6:12543–12557. 2015. View Article : Google Scholar : PubMed/NCBI
8	Su W, Xu M, Chen X, Nie L, Chen N, Gong J, Zhang M, Su Z, Huang L and Zhou Q: MiR200c targets IRS1 and suppresses prostate cancer cell growth. Prostate. 75:855–862. 2015. View Article : Google Scholar : PubMed/NCBI
9	Puhr M, Hoefer J, Schäfer G, Erb HH, Oh SJ, Klocker H, Heidegger I, Neuwirt H and Culig Z: Epithelial-to-mesenchymal transition leads to docetaxel resistance in prostate cancer and is mediated by reduced expression of miR-200c and miR-205. Am J Pathol. 181:2188–2201. 2012. View Article : Google Scholar : PubMed/NCBI
10	Radisky DC: miR-200c at the nexus of epithelial-mesenchymal transition, resistance to apoptosis, and the breast cancer stem cell phenotype. Breast Cancer Res. 13:1102011. View Article : Google Scholar : PubMed/NCBI
11	Qiu M, Liang Z, Chen L, Tan G, Liu L, Wang K, Chen H and Liu J: MicroRNA-200c suppresses cell growth and metastasis by targeting Bmi-1 and E2F3 in renal cancer cells. Exp Ther Med. 13:1329–1336. 2017. View Article : Google Scholar : PubMed/NCBI
12	Zhang H, Sun Z, Li Y, Fan D and Jiang H: MicroRNA-200c binding to FN1 suppresses the proliferation, migration and invasion of gastric cancer cells. Biomed Pharmacother. 88:285–292. 2017. View Article : Google Scholar : PubMed/NCBI
13	Chen P, Guo X, Zhang L, Zhang W, Zhou Q, Tian Z, Zheng Y, Liao Q, Wang H, Li G, et al: MiR-200c is a cMyc-activated miRNA that promotes nasopharyngeal carcinoma by downregulating PTEN. Oncotarget. 8:5206–5218. 2017.PubMed/NCBI
14	Zhang M, Nie L, Su Z, Xu M, Chen N, Gong J, Xie H, Zhong J, Tan J, Xu Y, et al: MicroRNA200c suppresses urothelial carcinoma invasiveness by targeting FSCN1. Int J Clin Exp Pathol. 10:5665–5674. 2017.
15	D'Ippolito E, Plantamura I, Bongiovanni L, Casalini P, Baroni S, Piovan C, Orlandi R, Gualeni AV, Gloghini A, Rossini A, et al: miR-9 and miR-200 regulate PDGFRβ-mediated endothelial differentiation of tumor cells in triple-negative breast cancer. Cancer Res. 76:5562–5572. 2016. View Article : Google Scholar : PubMed/NCBI
16	Shimono Y, Zabala M, Cho RW, Lobo N, Dalerba P, Qian D, Diehn M, Liu H, Panula SP, Chiao E, et al: Downregulation of miRNA-200c links breast cancer stem cells with normal stem cells. Cell. 138:592–603. 2009. View Article : Google Scholar : PubMed/NCBI
17	Gregory PA, Bert AG, Paterson EL, Barry SC, Tsykin A, Farshid G, Vadas MA, Khew-Goodall Y and Goodall GJ: The miR-200 family and miR-205 regulate epithelial to mesenchymal transition by targeting ZEB1 and SIP1. Nat Cell Biol. 10:593–601. 2008. View Article : Google Scholar : PubMed/NCBI
18	Tang X, Tang X, Gal J, Kyprianou N, Zhu H and Tang G: Detection of microRNAs in prostate cancer cells by microRNA array. Methods Mol Biol. 732:69–88. 2011. View Article : Google Scholar : PubMed/NCBI
19	Zha S, Ferdinandusse S, Denis S, Wanders RJ, Ewing CM, Luo J, De Marzo AM and Isaacs WB: Alpha-methylacyl-CoA racemase as an androgen-independent growth modifier in prostate cancer. Cancer Res. 63:7365–7376. 2003.PubMed/NCBI
20	Jindal Y, Singh A, Kumar R, Varma K, Misra V, Misra SP and Dwivedi M: Expression of alpha methylacyl CoA racemase (AMACR) in gastric adenocarcinoma and its correlation with helicobacter pylori infection. J Clin Diagn Res. 10:EC10–EC12. 2016.PubMed/NCBI
21	Zhang X, Leav I, Revelo MP, Deka R, Medvedovic M, Jiang Z and Ho SM: Deletion hotspots in AMACR promoter CpG island are cis-regulatory elements controlling the gene expression in the colon. PLoS Genet. 5:e10003342009. View Article : Google Scholar : PubMed/NCBI
22	Li CF, Chen LT, Lan J, Chou FF, Lin CY, Chen YY, Chen TJ, Li SH, Yu SC, Fang FM, et al: AMACR amplification and overexpression in primary imatinib-naïve gastrointestinal stromal tumors: A driver of cell proliferation indicating adverse prognosis. Oncotarget. 5:11588–11603. 2014.PubMed/NCBI
23	Li CF, Fang FM, Lan J, Wang JW, Kung HJ, Chen LT, Chen TJ, Li SH, Wang YH, Tai HC, et al: AMACR amplification in myxofibrosarcomas: A mechanism of overexpression that promotes cell proliferation with therapeutic relevance. Clin Cancer Res. 20:6141–6152. 2014. View Article : Google Scholar : PubMed/NCBI
24	Pinto F, Pértega-Gomes N, Vizcaíno JR, Andrade RP, Cárcano FM and Reis RM: Brachyury as a potential modulator of androgen receptor activity and a key player in therapy resistance in prostate cancer. Oncotarget. 7:28891–28902. 2016. View Article : Google Scholar : PubMed/NCBI
25	Chen W, Wu W, Zhao J, Yu C, Liu W, Jiang A and Zhang J: Molecular cloning and preliminary analysis of the human alpha-methylacyl-CoA racemase promoter. Mol Biol Rep. 36:423–430. 2009. View Article : Google Scholar : PubMed/NCBI
26	Zha S and Isaacs WB: A nonclassic CCAAT enhancer element binding protein binding site contributes to alpha-methylacyl-CoA racemase expression in prostate cancer. Mol Cancer Res. 3:110–118. 2005. View Article : Google Scholar : PubMed/NCBI
27	Chen N, Chen X, Huang R, Zeng H, Gong J, Meng W, Lu Y, Zhao F, Wang L and Zhou Q: BCL-xL is a target gene regulated by hypoxia-inducible factor-1{alpha}. J Biol Chem. 284:10004–10012. 2009. View Article : Google Scholar : PubMed/NCBI
28	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
29	Chen X, Gong J, Zeng H, Chen N, Huang R, Huang Y, Nie L, Xu M, Xia J, Zhao F, et al: MicroRNA145 targets BNIP3 and suppresses prostate cancer progression. Cancer Res. 70:2728–2738. 2010. View Article : Google Scholar : PubMed/NCBI
30	Ferdinandusse S, Denis S, IJlst L, Dacremont G, Waterham HR and Wanders RJ: Subcellular localization and physiological role of alpha-methylacyl-CoA racemase. J Lipid Res. 41:1890–1896. 2000.PubMed/NCBI
31	Mobley JA, Leav I, Zielie P, Wotkowitz C, Evans J, Lam YW, L'Esperance BS, Jiang Z and Ho SM: Branched fatty acids in dairy and beef products markedly enhance alpha-methylacyl-CoA racemase expression in prostate cancer cells in vitro. Cancer Epidemiol Biomarkers Prev. 12:775–783. 2003.PubMed/NCBI
32	Erdmann K, Kaulke K, Thomae C, Huebner D, Sergon M, Froehner M, Wirth MP and Fuessel S: Elevated expression of prostate cancer-associated genes is linked to down-regulation of microRNAs. BMC Cancer. 14:822014. View Article : Google Scholar : PubMed/NCBI
33	Festuccia C, Gravina GL, Mancini A, Muzi P, Cesare ED, Kirk R, Smith M, Hughes S, Gibson R, Lian LY, et al: Trifluoroibuprofen inhibits α-methylacyl coenzyme A racemase (AMACR/P504S), reduces cancer cell proliferation and inhibits in vivo tumor growth in aggressive prostate cancer models. Anticancer Agents Med Chem. 14:1031–1041. 2014. View Article : Google Scholar : PubMed/NCBI