Elsevier

Experimental Cell Research

Volume 348, Issue 2, 1 November 2016, Pages 190-200
Experimental Cell Research

Research Article
miR-1207-3p regulates the androgen receptor in prostate cancer via FNDC1/fibronectin

https://doi.org/10.1016/j.yexcr.2016.09.021Get rights and content

Highlights

  • Expression of microRNA-1207-3p is significantly lost in prostate cancer (PCa) cells.

  • MicroRNA-1207-3p regulates proliferation, apoptosis, and migration via direct molecular targeting of the 3′UTR of FNDC1.

  • MicroRNA-1207-3p regulates proliferation, apoptosis, and migration via direct molecular targeting of the 3′UTR of FNDC1.

  • FNDC1, FN1, and AR are concurrently overexpressed in metastatic PCa.

Abstract

Prostate cancer (PCa) is frequently diagnosed in men, and dysregulation of microRNAs is characteristic of many cancers. MicroRNA-1207-3p is encoded at the non-protein coding gene locus PVT1 on the 8q24 human chromosomal region, an established PCa susceptibility locus. However, the role of microRNA-1207-3p in PCa is unclear. We discovered that microRNA-1207-3p is significantly underexpressed in PCa cell lines in comparison to normal prostate epithelial cells. Increased expression of microRNA-1207-3p in PCa cells significantly inhibits proliferation, migration, and induces apoptosis via direct molecular targeting of FNDC1, a protein which contains a conserved protein domain of fibronectin (FN1). FNDC1, FN1, and the androgen receptor (AR) are significantly overexpressed in PCa cell lines and human PCa, and positively correlate with aggressive PCa. Prostate tumor FN1 expression in patients that experienced PCa-specific death is significantly higher than in patients that remained alive. Furthermore, FNDC1, FN1 and AR are concomitantly overexpressed in metastatic PCa. Consequently, these studies have revealed a novel microRNA-1207-3p/FNDC1/FN1/AR regulatory pathway in PCa.

Graphical abstract

miR-1207-3p/FNDC1/FN1/AR is a novel regulatory pathway in prostate cancer.

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Introduction

Prostate cancer (PCa) is the second most common cause of cancer-specific deaths in the U.S. accounting for approximately 13% of cancer-related deaths [1], [2], [3]. In 2014, it was estimated that the U.S. had 233,000 new cases of and 29,480 deaths from PCa [4], [5]. Established risk factors include familial genetics, and African ancestry [4], [6]. Thus, PCa is a multi-factorial, complex disease, with the exact mechanisms for its development and progression unclear. Understanding the molecular mechanisms underlying the development and progression of PCa is necessary. This will aid in the discovery of novel and efficacious biomarkers with applications in early PCa detection and molecular therapeutic targeting.

Genome-wide association studies (GWAS) have identified several loci and genetic variants that increase the risk of PCa. One of the most important susceptibility loci for PCa is the 8q24 human chromosomal region [7], [8], [9]. Recent studies demonstrated that PVT1, which is a 300 kb long non-protein coding gene locus found at the 8q24 human chromosomal region, is dysregulated in PCa [10], [11], [12], [13], [14]. The PVT1 gene locus encodes six microRNAs (miRNAs). Interestingly, five of the six microRNAs are not associated with annotated PVT1 exons, including microRNA-1207-3p (miR-1207-3p) [10], [15]. Furthermore, no known biological functions for this intron-derived miR-1207-3p has been previously reported.

Aberrant miRNA expression and function has potential applications in cancers. The objective of this study was to determine the expression, function, and molecular mechanisms of action of miR-1207-3p in PCa. Our study discovered a novel role of miR-1207-3p in the regulation of critical cellular functions in PCa via targeting of fibronectin type III domain containing 1 (FNDC1), leading to loss of fibronectin (FN1) expression and subsequent loss of androgen receptor (AR) expression. We also discovered that components of this novel molecular pathway, FNDC1, FN1, and AR are overexpressed in metastatic PCa. This discovery of a novel miR-1207-3p-dependent regulatory mechanism in PCa reveals the possibility of clinical applications for miR-1207-3p in PCa.

Section snippets

Cell culture

Detailed description is contained in Supplemental text. All cell lines have been tested and authenticated.

Transfection of oligonucleotide inhibitor and mimic of miR-1207-3p

Cells were seeded in 6-well plates. After reaching 60 to 70% confluence, media is replaced with Opti-MEM (Thermo Fisher Scientific Inc; Wilmington, DE, U.S.A) and cells are transfected with either a 50 nM non-targeting negative control oligonucleotide (MISSION® Synthetic microRNA Negative Control, product# NCSTUD001), 50 nM miR-1207-3p oligonucleotide mimic (MISSION® microRNA Mimic, product#

miR-1207-3p is significantly underexpressed in PCa cells

To investigate the role of miR-1207-3p in PCa, a panel of 8 prostate cell lines modeling different clinical characteristics of PCa was used. The panel included WPE1-NA22 (derived from RWPE-1, indolent, androgen-dependent, from Caucasian male), MDA PCa 2b (aggressive, androgen-dependent, from Black male), PC-3 (aggressive, androgen-independent, from Caucasian male), E006AA (indolent, androgen-independent, from Black male), E006AA-hT (derived from E006AA, aggressive, androgen-independent, from

Discussion

Prostate cancer (PCa) is the most frequently diagnosed solid organ cancer in males in the developed world [36]. Even though PCa is so prevalent, the molecular mechanisms underlying its development and progression remain largely unclear. Increasing evidence has demonstrated that many miRNAs are aberrantly expressed in cancers, suggesting that variations in miRNA expression are common events in tumorigenesis [37], [38], [39]. These small non-coding RNAs are attractive for clinical applications

Conclusions

In conclusion, we identified loss of miR-1207-3p expression in PCa cells, and demonstrate that miR-1207-3p is an important regulator of proliferation, apoptosis, and migration of PCa cells. Importantly, we describe for the first time the miR-1207-3p/FNDC1/FN1/AR novel regulatory pathway. And we report, for the first time, clinical data indicating that this novel molecular pathway may be important in metastatic PCa. Consequently, this novel miR-1207-3p-dependent pathway may be actionable for

Author contributions

DD wrote the main manuscript and prepared Figs. 1–6. AI prepared Fig. 1b and reviewed the manuscript. JYP prepared Table 1 and corresponding method section. TA and KK prepared Supplementary Fig. S3 and corresponding method section. BDR prepared Fig. 5 and corresponding method section. JRO made intellectual contributions and reviewed the manuscript. OOO supervised all the work from conception to manuscript preparation and review. All authors reviewed the manuscript.

Disclosure of potential conflicts of interest

The authors declare no potential conflicts of interests.

Acknowledgements

We would like to thank Jeannette Huaman, Jazmine Joseph, Victoria Durojaiye and Tamara Gillot for their technical assistance. We would also like to thank Jyoti Chouhan for reading our manuscript and providing helpful comments. Work in Dr. Ogunwobi's laboratory is supported by the NIMHD/NIH grant to Hunter College: 8 G12 MD007599.

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