Biopolym. Cell. 2017; 33(5):335-355.
Genomics, Transcriptomics and Proteomics
Expression of epithelial-mesenchymal transition-related genes in prostate tumours
1Gerashchenko G. V., 1Mankovska O. S., 2Dmitriev A. A., 1Mevs L. V., 1Rosenberg E. E., 3Pikul M. V., 3Marynychenko M. V., 4Gryzodub O. P., 3Stakhovsky E. O., 1Kashuba V. I.
  1. Institute of Molecular Biology and Genetics, NAS of Ukraine
    150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680
  2. Engelhardt Institute of Molecular Biology, Russian Academy of Sciences
    32, Vavilova Str., Moscow, Russian Federation, 119991
  3. National Cancer Institute
    33/43, Lomonosova Str., Kyiv, Ukraine, 03022
  4. State Institution «Institute of Urology of NAMS of Ukraine»
    9-a, Yu. Kotsubyns'koho Str., Kyiv, Ukraine, 04053

Abstract

Aim. To detect expression of EMT-related genes in prostate tumor samples and analyze a possible correlation between gene expression level and clinical characteristics of prostate cancer in different groups. Methods. Expression of 19 genes was analyzed in 37 frozen samples of prostate cancer tissues at different tumor stages and Gleason scores, 37 paired conventionally normal prostate tissues and 20 samples of prostate adenomas, using quantitative PCR. Results. We have found that nine genes were expressed differently in benign and malignant prostate tumors, namely AR (1 isof), AR (2 isof), PTEN, VIM, MMP9, KRT18, PCA3, HOTAIR and SCHLAP1. When different tumor stages were compared, we could identify six differentially expressed genes: KRT18, MMP9, VIM, PCA3, HOTAIR and SCHLAP1; when samples of tumors with different Gleason score were compared, we found that eight genes were expressed differently: AR (1isof), CDH1, KRT18, MMP9, OCLN, PCA3, HOTAIR and SCHLAP1. The data had a high level of heterogeneity pottentially due to various molecular subtypes of prostate cancer, i.e. a luminal subtype with ahigh expression of CDH1, OCLN, AR(1 isof), KRT18, NKX3-1 and PSA; the stem-like subtype with a high expression of mesenchymal markers CDH2, FN1 and VIM and low expression of the epithelial markers. It is noteworthy that lncRNAs were specifically expressed in these two molecular subtypes. Conclusions. EMT-related genes were differentially expressed in benign and malignant prostate tumors. High heterogeneity of expression levels, especially in adenocarcinoma groups, might suggest the existence of at least two different molecular subtypes, luminal and stem-like. Further experiments are necessary for specification of the molecular subtypes of prostate adenocarcinoma.
Keywords: prostate tumors, EMT, relative gene expression, molecular subtypes, lncRNA

References

[1] Nguyen T, Boldt RG, Rodrigues G. Prognostic factors for prostate cancer endpoints following biochemical failure: a review of the literature. Cureus. 2015; 7(1):e238.
[2] Gerlinger M, Catto JW, Orntoft TF, Real FX, Zwarthoff EC, Swanton C. Intratumour heterogeneity in urologic cancers: from molecular evidence to clinical implications. Eur Urol. 2015;67(4):729-37. doi:
[3] Tomaskovic-Crook E, Thompson EW, Thiery JP. Epithelial to mesenchymal transition and breast cancer. Breast Cancer Res. 2009;11(6):213. doi:
[4] Thiery JP. Epithelial-mesenchymal transitions in tumour progression. Nat Rev Cancer. 2002;2(6):442-54.
[5] Chappell WH, Abrams SL, Lertpiriyapong K, Fitzgerald TL, Martelli AM, Cocco L, Rakus D, Gizak A, Terrian D, Steelman LS, McCubrey JA. Novel roles of androgen receptor, epidermal growth factor receptor, TP53, regulatory RNAs, NF-kappa-B, chromosomal translocations, neutrophil associated gelatinase, and matrix metalloproteinase-9 in prostate cancer and prostate cancer stem cells. Adv Biol Regul. 2016;60:64-87. doi:
[6] Song H, Zhang B, Watson MA, Humphrey PA, Lim H, Milbrandt J. Loss of Nkx3.1 leads to the activation of discrete downstream target genes during prostate tumorigenesis. Oncogene. 2009;28(37):3307-19. doi:
[7] Pećina-Slaus N. Tumor suppressor gene E-cadherin and its role in normal and malignant cells. Cancer Cell Int. 2003;3(1):17.
[8] Martin TA, Mansel RE, Jiang WG. Loss of occludin leads to the progression of human breast cancer. Int J Mol Med. 2010;26(5):723-34.
[9] Niknami Z, Eslamifar A, Emamirazavi A, Ebrahimi A, Shirkoohi R. The association of vimentin and fibronectin gene expression with epithelial-mesenchymal transition and tumor malignancy in colorectal carcinoma. EXCLI J. 2017;16:1009-1017.
[10] Zhong WD, Han ZD, He HC, Bi XC, Dai QS, Zhu G, Ye YK, Liang YX, Qin WJ, Zhang Z, Zeng GH, Chen ZN. CD147, MMP-1, MMP-2 and MMP-9 protein expression as significant prognostic factors in human prostate cancer. Oncology. 2008;75(3-4):230-6. doi:
[11] Zhu ML, Kyprianou N. Role of androgens and the androgen receptor in epithelial-mesenchymal transition and invasion of prostate cancer cells. FASEB J. 2010;24(3):769-77. doi:
[12] Prensner JR, Rubin MA, Wei JT, Chinnaiyan AM. Beyond PSA: the next generation of prostate cancer biomarkers. Sci Transl Med. 2012;4(127):127rv3. doi:
[13] Fortier AM, Asselin E, Cadrin M. Keratin 8 and 18 loss in epithelial cancer cells increases collective cell migration and cisplatin sensitivity through claudin1 up-regulation. J Biol Chem. 2013;288(16):11555-71. doi: PubMed Central
[14] Ha SA, Lee YS, Kim HK, Yoo J, Kim S, Gong GH, Lee YK, Kim JW. The prognostic potential of keratin 18 in breast cancer associated with tumor dedifferentiation, and the loss of estrogen and progesterone receptors. Cancer Biomark. 2011;10(5):219-31. doi:
[15] Tretiakova MS, Wei W, Boyer HD, Newcomb LF, Hawley S, Auman H, Vakar-Lopez F, McKenney JK, Fazli L, Simko J, Troyer DA, Hurtado-Coll A, Thompson IM Jr, Carroll PR, Ellis WJ, Gleave ME, Nelson PS, Lin DW, True LD, Feng Z, Brooks JD. Prognostic value of Ki67 in localized prostate carcinoma: a multi-institutional study of >1000 prostatectomies. Prostate Cancer Prostatic Dis. 2016;19(3):264-70. doi:
[16] O'Donovan N, Crown J, Stunell H, Hill AD, McDermott E, O'Higgins N, Duffy MJ. Caspase 3 in breast cancer. Clin Cancer Res. 2003;9(2):738-42.
[17] Jäger R, Zwacka RM. The enigmatic roles of caspases in tumor development. Cancer (Basel)s. 2010; 2(4):1952-1979.
[18] Obexer P, Ausserlechner MJ. X-linked inhibitor of apoptosis protein - a critical death resistance regulator and therapeutic target for personalized cancer therapy. Front Oncol. 2014;4:197. doi:
[19] Lemos AE, Ferreira LB, Batoreu NM, de Freitas PP, Bonamino MH, Gimba ER. PCA3 long noncoding RNA modulates the expression of key cancer-related genes in LNCaP prostate cancer cells. Tumour Biol. 2016;37(8):11339-48. doi:
[20] Hajjari M, Salavaty A. HOTAIR: an oncogenic long non-coding RNA in different cancers. Cancer Biol Med. 2015; 12(1):1-9. doi:
[21] Prensner JR, Iyer MK, Sahu A, Asangani IA, Cao Q, Patel L, Vergara IA, Davicioni E, Erho N, Ghadessi M, Jenkins RB, Triche TJ, Malik R, Bedenis R, McGregor N, Ma T, Chen W, Han S, Jing X, Cao X, Wang X, Chandler B, Yan W, Siddiqui J, Kunju LP, Dhanasekaran SM, Pienta KJ, Feng FY, Chinnaiyan AM. The long noncoding RNA SChLAP1 promotes aggressive prostate cancer and antagonizes the SWI/SNF complex. Nat Genet. 2013;45(11):1392-8. doi:
[22] Schmidt U, Fuessel S, Koch R, Baretton GB, Lohse A, Tomasetti S, Unversucht S, Froehner M, Wirth MP, Meye A. Quantitative multi-gene expression profiling of primary prostate cancer. Prostate. 2006;66(14):1521-34.
[23] Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 2001;25(4):402-8.
[24] Benjamini Y, Hochberg Y. Controlling the false discovery rate: a practical and powerful approach to multiple testing. J R Stat Soc Series B Stat Methodol. 1995, 57(1): 289-300.
[25] Montanari M, Rossetti S, Cavaliere C, D'Aniello C, Malzone MG, Vanacore D, Di Franco R, La Mantia E, Iovane G, Piscitelli R, Muscariello R, Berretta M, Perdonà S, Muto P, Botti G, Bianchi AAM, Veneziani BM, Facchini G. Epithelial-mesenchymal transition in prostate cancer: an overview. Oncotarget. 2017;8(21):35376-35389. doi:
[26] Zhao SG, Chang SL, Erho N, Yu M, Lehrer J, Alshalalfa M, Speers C, Cooperberg MR, Kim W, Ryan CJ, Den RB, Freedland SJ, Posadas E, Sandler H, Klein EA, Black P, Seiler R, Tomlins SA, Chinnaiyan AM, Jenkins RB, Davicioni E, Ross AE, Schaeffer EM, Nguyen PL, Carroll PR, Karnes RJ, Spratt DE, Feng FY. Associations of luminal and basal subtyping of prostate cancer with prognosis and response to androgen deprivation therapy. JAMA Oncol. 2017;3(12):1663-1672. doi:
[27] Park JW, Lee JK, Phillips JW, Huang P, Cheng D, Huang J, Witte ON. Prostate epithelial cell of origin determines cancer differentiation state in an organoid transformation assay. Proc Natl Acad Sci U S A. 2016;113(16):4482-7. doi:
[28] Yang Z, Yu L, Wang Z. PCA3 and TMPRSS2-ERG gene fusions as diagnostic biomarkers for prostate cancer. Chin J Cancer Res. 2016;28(1):65-71. doi:
[29] Prensner JR, Zhao S, Erho N, Schipper M, Iyer MK, Dhanasekaran SM, Magi-Galluzzi C, Mehra R, Sahu A, Siddiqui J, Davicioni E, Den RB, Dicker AP, Karnes RJ, Wei JT, Klein EA, Jenkins RB, Chinnaiyan AM, Feng FY. RNA biomarkers associated with metastatic progression in prostate cancer: a multi-institutional high-throughput analysis of SChLAP1. Lancet Oncol. 2014;15(13):1469-80. doi:
[30] Ahrens-Fath I, Politz O, Geserick C, Haendler B. Androgen receptor function is modulated by the tissue-specific AR45 variant. FEBS J. 2005;272(1):74-84.
[31] Smith BA, Sokolov A, Uzunangelov V, Baertsch R, Newton Y, Graim K, Mathis C, Cheng D, Stuart JM, Witte ON. A basal stem cell signature identifies aggressive prostate cancer phenotypes. Proc Natl Acad Sci U S A. 2015;112(47):E6544-52. doi:
[32] Deng J, Yang M, Jiang R, An N, Wang X, Liu B. Long Non-Coding RNA HOTAIR Regulates the Proliferation, Self-Renewal Capacity, Tumor Formation and Migration of the Cancer Stem-Like Cell (CSC) Subpopulation Enriched from Breast Cancer Cells. PLoS One. 2017;12(1):e0170860. doi:
[33] Cancer Genome Atlas Research Network. The Molecular Taxonomy of Primary Prostate Cancer. Cell. 2015;163(4):1011-25. doi: