A multimarker qPCR platform for the characterisation of endometrial cancer

Supernat,Anna; Łapińska-Szumczyk,Sylwia; Majewska,Hanna; Gulczyński,Jacek; Biernat,Wojciech; Wydra,Dariusz; Żaczek,Anna J.

doi:10.3892/or.2013.2924

A multimarker qPCR platform for the characterisation of endometrial cancer

Authors:
- Anna Supernat
- Sylwia Łapińska-Szumczyk
- Hanna Majewska
- Jacek Gulczyński
- Wojciech Biernat
- Dariusz Wydra
- Anna J. Żaczek
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Affiliations: Department of Medical Biotechnology, Intercollegiate Faculty of Biotechnology, University of Gdańsk and Medical University of Gdańsk, Dębinki 1, 80-211 Gdańsk, Poland, Department of Gynaecology, Gynaecological Oncology and Gynaecological Endocrinology, Medical University of Gdańsk, Kliniczna 1a, 80-402 Gdańsk, Poland, Department of Pathomorphology, Medical University of Gdańsk, Dębinki 7, 80-211 Gdańsk, Poland, Department of Pathology and Neuropathology, Medical University of Gdańsk, Dębinki 1, 80-211 Gdańsk, Poland
Published online on: December 13, 2013 https://doi.org/10.3892/or.2013.2924
Pages: 1003-1013

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Abstract

The molecular background of endometrial cancer (EC) has not been fully elucidated. In the present study, we developed a quantitative PCR (qPCR) platform to examine the gene dosages of the potential molecular markers MGB1, TOP2A, ERBB1-4, MYC, CCND1, ESR1 and PI3K. The platform was applied in samples collected from 157 EC patients (stage I-IV) to verify its clinical utility and to examine the diagnostic and prognostic significance of the analysed biomarkers. The gene dosage pattern of the ERBB family and its downstream effectors PI3K and MYC showed particularly strong correlations with clinicopathological data. The ERBB PI3K/Akt pathway was upregulated in 31 (20%) of 156 cases. Activation of the ERBB PI3K/Akt pathway was positively correlated with a higher stage (p=0.001), higher grade (p=0.001), histological type II disease (p=0.0003) and metastases (p=0.02). The implemented hierarchical clustering revealed that cluster 2 was characterised by high copy numbers of the studied genes. Cluster 2 was associated with shorter overall survival (p=0.05). The platform was found to be a fast and simple method for direct analysis of the genes involved in uterine carcinogenesis, making it feasible for EC biology characterisation.

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1	Mirantes C, Espinosa I, Ferrer I, Dolcet X, Prat J and Matias-Guiu X: Epithelial-to-mesenchymal transition and stem cells in endometrial cancer. Hum Pathol. 44:1973–1981. 2013. View Article : Google Scholar : PubMed/NCBI
2	Wojciechowska U and Didkowska J: Cancer in Poland in 2010. Nowotwory. 63:197–216. 2013.
3	Bray F, Dos Santos Silva I, Moller H and Weiderpass E: Endometrial cancer incidence trends in Europe: underlying determinants and prospects for prevention. Cancer Epidemiol Biomarkers Prev. 14:1132–1142. 2005. View Article : Google Scholar : PubMed/NCBI
4	Bokhman JV: Two pathogenetic types of endometrial carcinoma. Gynecol Oncol. 15:10–17. 1983. View Article : Google Scholar : PubMed/NCBI
5	Horn LC, Meinel A, Handzel R and Einenkel J: Histopathology of endometrial hyperplasia and endometrial carcinoma: an update. Ann Diagn Pathol. 11:297–311. 2007. View Article : Google Scholar
6	Amant F, Moerman P, Neven P, Timmerman D, Van Limbergen E and Vergote I: Endometrial cancer. Lancet. 366:491–505. 2005. View Article : Google Scholar : PubMed/NCBI
7	Randall TC and Kurman RJ: Progestin treatment of atypical hyperplasia and well-differentiated carcinoma of the endometrium in women under age 40. Obstet Gynecol. 90:434–440. 1997. View Article : Google Scholar : PubMed/NCBI
8	Sherman ME, Sturgeon S, Brinton LA, et al: Risk factors and hormone levels in patients with serous and endometrioid uterine carcinomas. Mod Pathol. 10:963–968. 1997.PubMed/NCBI
9	Ayhan A, Gultekin M and Dursun P: Textbook of Gynaecological Oncology. Günes Publishing; Ankara: 2010
10	Yeramian A, Moreno-Bueno G, Dolcet X, et al: Endometrial carcinoma: molecular alterations involved in tumor development and progression. Oncogene. 32:403–413. 2012. View Article : Google Scholar : PubMed/NCBI
11	Busmanis I, Ho TH, Tan SB and Khoo KS: p53 and bcl-2 expression in invasive and pre-invasive uterine papillary serous carcinoma and atrophic endometrium. Ann Acad Med Singapore. 34:421–425. 2005.PubMed/NCBI
12	Caduff RF, Johnston CM and Frank TS: Mutations of the Ki-ras oncogene in carcinoma of the endometrium. Am J Pathol. 146:182–188. 1995.PubMed/NCBI
13	Mutter GL, Lin MC, Fitzgerald JT, et al: Altered PTEN expression as a diagnostic marker for the earliest endometrial precancers. J Natl Cancer Inst. 92:924–930. 2000. View Article : Google Scholar : PubMed/NCBI
14	Mutter GL, Boynton KA, Faquin WC, Ruiz RE and Jovanovic AS: Allelotype mapping of unstable microsatellites establishes direct lineage continuity between endometrial precancers and cancer. Cancer Res. 56:4483–4486. 1996.
15	Strissel PL, Ellmann S, Loprich E, et al: Early aberrant insulin-like growth factor signaling in the progression to endometrial carcinoma is augmented by tamoxifen. Int J Cancer. 123:2871–2879. 2008. View Article : Google Scholar : PubMed/NCBI
16	Rose PG: Endometrial carcinoma. N Engl J Med. 335:640–649. 1996. View Article : Google Scholar : PubMed/NCBI
17	Onuma K, Dabbs DJ and Bhargava R: Mammaglobin expression in the female genital tract: immunohistochemical analysis in benign and neoplastic endocervix and endometrium. Int J Gynecol Pathol. 27:418–425. 2008. View Article : Google Scholar : PubMed/NCBI
18	Grushko TA, Filiaci VL, Mundt AJ, et al: An exploratory analysis of HER-2 amplification and overexpression in advanced endometrial carcinoma: a gynecologic oncology group study. Gynecol Oncol. 108:3–9. 2008. View Article : Google Scholar : PubMed/NCBI
19	Zaczek A, Wełnicka-Jaśkiewicz M, Bielawski KP, et al: Gene copy numbers of HER family in breast cancer. J Cancer Res Clin Oncol. 134:271–279. 2008. View Article : Google Scholar
20	Zaczek A, Brandt B and Bielawski KP: The diverse signaling network of EGFR, HER2, HER3 and HER4 tyrosine kinase receptors and the consequences for therapeutic approaches. Histol Histopathol. 20:1005–1015. 2005.PubMed/NCBI
21	Salvesen HB, Carter SL, Mannelqvist M, et al: Integrated genomic profiling of endometrial carcinoma associates aggressive tumors with indicators of PI3 kinase activation. Proc Natl Acad Sci USA. 106:4834–4839. 2009. View Article : Google Scholar : PubMed/NCBI
22	Ponzielli R, Katz S, Barsyte-Lovejoy D and Penn LZ: Cancer therapeutics: targeting the dark side of Myc. Eur J Cancer. 41:2485–2501. 2005. View Article : Google Scholar : PubMed/NCBI
23	Semczuk A and Jakowicki JA: Alterations of pRb1-cyclin D1-cdk4/6-p16^INK4A pathway in endometrial carcinogenesis. Cancer Lett. 203:1–12. 2004. View Article : Google Scholar : PubMed/NCBI
24	Tsiambas E, Alexopoulou D, Lambropoulou S, Gerontopoulos K, Karakitsos P and Karameris A: Targeting topoisomerase IIa in endometrial adenocarcinoma: a combined chromogenic in situ hybridization and immunohistochemistry study based on tissue microarrays. Int J Gynecol Cancer. 16:1424–1431. 2006. View Article : Google Scholar
25	Pecorelli S: Revised FIGO staging for carcinoma of the vulva, cervix, and endometrium. Int J Gynaecol Obstet. 105:103–104. 2009. View Article : Google Scholar : PubMed/NCBI
26	Consultation WE: Appropriate body-mass index for Asian populations and its implications for policy and intervention strategies. Lancet. 363:157–163. 2004. View Article : Google Scholar : PubMed/NCBI
27	Jönsson G, Staaf J, Olsson E, et al: High-resolution genomic profiles of breast cancer cell lines assessed by tiling BAC array comparative genomic hybridization. Genes Chromosomes Cancer. 46:543–558. 2007.PubMed/NCBI
28	Vandesompele J, De Preter K, Pattyn F, et al: Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol. 3:research00342002. View Article : Google Scholar : PubMed/NCBI
29	Hellemans J, Mortier G, De Paepe A, Speleman F and Vandesompele J: qBase relative quantification framework and software for management and automated analysis of real-time quantitative PCR data. Genome Biol. 8:R192007. View Article : Google Scholar : PubMed/NCBI
30	McShane LM, Altman DG, Sauerbrei W, et al: REporting recommendations for tumor MARKer prognostic studies (REMARK). Nat Clin Pract Oncol. 2:416–422. 2005. View Article : Google Scholar
31	Järvinen TA, Tanner M, Rantanen V, et al: Amplification and deletion of topoisomerase IIα associate with ErbB-2 amplification and affect sensitivity to topoisomerase II inhibitor doxorubicin in breast cancer. Am J Pathol. 156:839–847. 2000.
32	Supernat A, Lapińska-Szumczyk S, Sawicki S, Wydra D, Biernat W and Zaczek AJ: Deregulation of RAD21 and RUNX1 expression in endometrial cancer. Oncol Lett. 4:727–732. 2012.
33	Inaki K and Liu ET: Structural mutations in cancer: mechanistic and functional insights. Trends Genet. 28:550–559. 2012. View Article : Google Scholar : PubMed/NCBI
34	Watson MA and Fleming TP: Mammaglobin, a mammary-specific member of the uteroglobin gene family, is overexpressed in human breast cancer. Cancer Res. 56:860–865. 1996.PubMed/NCBI
35	Classen-Linke I, Moss S, Gröting K, Beier HM, Alfer J and Krusche CA: Mammaglobin 1: not only a breast-specific and tumour-specific marker, but also a hormone-responsive endometrial protein. Histopathology. 61:955–965. 2012. View Article : Google Scholar : PubMed/NCBI
36	Raeder MB, Birkeland E, Trovik J, et al: Integrated genomic analysis of the 8q24 amplification in endometrial cancers identifies ATAD2 as essential to MYC-dependent cancers. PLoS One. 8:e548732013. View Article : Google Scholar : PubMed/NCBI
37	Esteller M, García A, Martínez i Palones JM, Cabero A and Reventós J: Detection of c-erbB-2/neu and fibroblast growth factor-3/INT-2 but not epidermal growth factor receptor gene amplification in endometrial cancer by differential polymerase chain reaction. Cancer. 75:2139–2146. 1995. View Article : Google Scholar
38	Liang H, Cheung LW, Li J, et al: Whole-exome sequencing combined with functional genomics reveals novel candidate driver cancer genes in endometrial cancer. Genome Res. 22:2120–2129. 2012. View Article : Google Scholar
39	Saghir FS, Rose IM, Dali AZ, Shamsuddin Z, Jamal AR and Mokhtar NM: Gene expression profiling and cancer-related pathways in type I endometrial carcinoma. Int J Gynecol Cancer. 20:724–731. 2010. View Article : Google Scholar : PubMed/NCBI
40	Konecny GE, Santos L, Winterhoff B, et al: HER2 gene amplification and EGFR expression in a large cohort of surgically staged patients with nonendometrioid (type II) endometrial cancer. Br J Cancer. 100:89–95. 2009. View Article : Google Scholar : PubMed/NCBI
41	Rolitsky CD, Theil KS, McGaughy VR, Copeland LJ and Niemann TH: HER-2/neu amplification and overexpression in endometrial carcinoma. Int J Gynecol Pathol. 18:138–143. 1999. View Article : Google Scholar : PubMed/NCBI
42	Tsikouras P, Bouchlariotou S, Vrachnis N, et al: Endometrial cancer: molecular and therapeutic aspects. Eur J Obstet Gynecol Reprod Biol. 169:1–9. 2013. View Article : Google Scholar : PubMed/NCBI
43	Fleming GF, Sill MW, Darcy KM, et al: Phase II trial of trastuzumab in women with advanced or recurrent, HER2-positive endometrial carcinoma: a Gynecologic Oncology Group study. Gynecol Oncol. 116:15–20. 2010. View Article : Google Scholar
44	Santin AD, Bellone S, Roman JJ, McKenney JK and Pecorelli S: Trastuzumab treatment in patients with advanced or recurrent endometrial carcinoma overexpressing HER2/neu. Int J Gynaecol Obstet. 102:128–131. 2008. View Article : Google Scholar : PubMed/NCBI
45	Santin AD: Letter to the Editor referring to the manuscript entitled: ‘Phase II trial of trastuzumab in women with advanced or recurrent HER-positive endometrial carcinoma: a Gynecologic Oncology Group study’ recently reported by Fleming et al, (Gynecol Oncol., 116;15–20;2010). Gynecol Oncol. 118:95–96; author reply 96–97. 2010.