Analysis of PMEPA1 Isoforms (a and b) as Selective Inhibitors of Androgen and TGF-β Signaling Reveals Distinct Biological and Prognostic Features in Prostate Cancer
Abstract
:1. Introduction
2. Results
2.1. Structures, Expressions and Distinct Regulations of PMEPA1 Isoforms in Prostate Cancer Cells
2.2. PMEPA1-a and -b Isoform mRNA Levels as a Biomarker for Prostate Cancer Aggressiveness
2.3. The Increased Ratio between Transcripts of PMEPA1-a and PMEPA1-b Indicated a Higher Gleason Score and More Aggressive Prostate Cancer
2.4. PMEPA1-a Promoted the Growth of AR Negative Prostate Cancer Cells and Inhibited TGF-β Signaling
2.5. PMEPA1-b Inhibited the Growth of Hormone Dependent Prostate Cancer Cells through Facilitating AR Protein Degradation and Blocking Androgen Signaling
3. Discussion
4. Materials and Methods
4.1. Prostate Cancer Specimens and Clinico-Pathological Data
4.2. RNA Isolation and Quantitative RT-PCR
4.3. The TCGA RNA Sequencing (RNA Seq) Data Acquisition
4.4. Analyses of the Correlation between PMEPA1 Isoform Expression Levels and Clinical Outcome Including Gleason Scores in Prostate Cancer Patients from the TCGA Dataset
4.5. Cell Culture
4.6. Immunoblotting
4.7. Plasmids and siRNAs
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
DoD Disclaimer
References
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PMEPA1-a | Forward | 5′-GCAACTG CAAACGCTCTTTGT-3′ |
PMEPA1-a | Reverse | 5′-GGACCGTGCAGACAGCTTGTA-3′ |
PMEPA1-a | probe | 6FAM-CATGGAGAT CACGGAGC-TAMRA |
PMEPA1-b | Forward | 5′-CATCATCCCCGAGCTGCT-3′ |
PMEPA1-b | Reverse | 5′- TGATCTGAACAAACTCCAGCTCC-3′ |
PMEPA1-b | probe | 6FAM-AGGCGGACAGTCTCCTGCGAAA-TAMRA |
KLK3/(PSA) | Forward | 5′-CCC ACT GCA TCA GGA ACA AA-3′ |
KLK3/(PSA) | Reverse | 5′- GAG CGG GTG TGG GAA GCT-3′ |
KLK3/(PSA) | probe | 6FAM- ACA CAG GCC AGG TAT TTC AGG TCA GCC -TAMRA |
AR | Forward | 5′- GGTGTCACTATGGAGCTCTCACAT -3′ |
AR | Reverse | 5′- GCAATCATTTCTGCTGGCG -3′ |
AR | probe | FAM- CTTCAAAAGAGCCGCTGAAGGGAAACAG –TAMRA |
NEDD9 | Forward | 5′-ATCAGCTGAGCCAGTTCCAG-3′ |
NEDD9 | Reverse | 5′-TGGGTCTCACATTGGTCAT-3′ |
NEDD9 | probe | 6FAM-AAGCCCTCTCAGAGCCTACC–TAMRA |
COLA1 | Forward | 5′-CAGGTCTCGGTCATGGTACCT-3′ |
COLA1 | Reverse | 5′-GTCGAGGGCCAAGACGAA-3′ |
COLA1 | probe | 6FAM-CATCCCACCAATCACCTGCGTACAGA-TAMRA |
THBS1 | Forward | 5′-TTGTCTTTGGAACCACACCA-3′ |
THBS1 | Reverse | 5′-TTGTCAAGGGTGAGGAGGAC-3′ |
THBS1 | probe | 6FAM-TGCAGCTATCAACAGTCCATTCCTCG -TAMRA |
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Sharad, S.; Sztupinszki, Z.M.; Chen, Y.; Kuo, C.; Ravindranath, L.; Szallasi, Z.; Petrovics, G.; Sreenath, T.L.; Dobi, A.; Rosner, I.L.; et al. Analysis of PMEPA1 Isoforms (a and b) as Selective Inhibitors of Androgen and TGF-β Signaling Reveals Distinct Biological and Prognostic Features in Prostate Cancer. Cancers 2019, 11, 1995. https://doi.org/10.3390/cancers11121995
Sharad S, Sztupinszki ZM, Chen Y, Kuo C, Ravindranath L, Szallasi Z, Petrovics G, Sreenath TL, Dobi A, Rosner IL, et al. Analysis of PMEPA1 Isoforms (a and b) as Selective Inhibitors of Androgen and TGF-β Signaling Reveals Distinct Biological and Prognostic Features in Prostate Cancer. Cancers. 2019; 11(12):1995. https://doi.org/10.3390/cancers11121995
Chicago/Turabian StyleSharad, Shashwat, Zsófia M. Sztupinszki, Yongmei Chen, Claire Kuo, Lakshmi Ravindranath, Zoltan Szallasi, Gyorgy Petrovics, Taduru L. Sreenath, Albert Dobi, Inger L. Rosner, and et al. 2019. "Analysis of PMEPA1 Isoforms (a and b) as Selective Inhibitors of Androgen and TGF-β Signaling Reveals Distinct Biological and Prognostic Features in Prostate Cancer" Cancers 11, no. 12: 1995. https://doi.org/10.3390/cancers11121995