Investigation of The Transcription Stages of Prostaglandin Endoperoxid H Synthase 2 (PTGS2), Calreticulin (CALR) and Keratin-19 (KRT19) Genes in the Breast Cancer MDA-MB-231

Abstract

Background: In this study, we aimed to determine the gene expressions of prostaglandin endoperoxide H synthase 2 (PTGS2), calreticulin (CALR), and keratin-19 (KRT19) genes at transcriptional level in MDA-MB-231 breast cancer cell line.
Materials and Methods: RNA was isolated from MDA-MB-231 breast cancer cells grown in culture medium, followed by cDNA synthesis. Real-time PCR with specific primers for PTGS2, CALR and KRT19 genes was used to determine the expression of these genes at the transcriptional level.
Results: In the MDA-MB-231 cell line, a 14.92-fold increase in PTGS2 gene expression, a 1.45-fold increase in CALR gene expression and a 6.72-fold increase in expression of the KRT19 gene were found.
Conclusions: It is predicted that the KRT19, CALR, PTGS2 genes, which are known to play a role in biological processes such as apoptosis resistance, metastasis, and angiogenesis in different solid cancers, may also play a role in the development of breast cancer and will have prognostic importance with further studies.

Key words: Breast cancer, PTGS2 gene, CALR gene, KRT19 gene

MDA-MB-231 Meme Kanseri Hücre Dizisinde Prostoglandin Endoperoksid H Sentaz 2 (PTGS2), Kalretikulin (CALR) ve Keratin-19 (KRT19) Genlerinin Transkripsiyon Aşamasında Anlatımlarının Araştırılması

Abstract

Amaç: Bu çalışmada MDA-MB-231 meme kanser hücre hattında prostaglandin endoperoksid H sentaz 2 (PTGS2), kalretikulin (CALR) ve keratin-19 (KRT19) genlerinin transkripsiyon düzeyindeki gen anlatımlarının belirlenmesi amaçlanmıştır.
Materyal ve metod: Kültür ortamında çoğaltılan MDA-MB-231 meme kanser hücrelerinden RNA izolasyonu yapılmış ardından cDNA sentezi gerçekleştirilmiştir. PTGS2, CALR ve KRT19 genlerine spesifik primerler ile eş zamanlı PCR yapılarak, bu genlerin ifadesi transkripsiyonel seviyede belirlenmiştir.
Bulgular: MDA-MB-231 hücre hattında PTGS2 gen ifadesinde 14,92 kat; CALR gen ifadesinde 1,45 kat; KRT19 geninin ifadesinde ise 6,72 kat artış olduğu saptanmıştır.
Sonuç: Farklı solid kanserlerde, apoptoz direnci, metastaz, anjiyogenez gibi biyolojik süreçlerde rol aldığı bilinen KRT19, CALR, PTGS2 genlerinin meme kanseri gelişiminde de rol alabileceği ve ileride yapılacak detaylı çalışmalarla prognostik öneme sahip olacağı ön görülmektedir.

Anahtar Kelimeler: Meme kanseri, PTGS2 geni, CALR geni, KRT19 geni

Keywords

• Meme kanseri
• PTGS2 geni
• CALR geni
• KRT19 geni

References

1. Referans1.Siegel RL, Miller KD, Jemal A. Cancer Statistics. CA Cancer J Clin 2017; 67: 7-30
2. Referans2.Benson JR, Jatoi I. The global breast cancer burden. Future Oncol 2012;8(6):697–702
3. Referans3.Mavaddat N, Pharoah PD, Michailidou K, Tyrer J, Brook MN, Bolla MK, et al. Prediction of breast cancer risk based on profiling with common genetic variants. J Natl Cancer Inst. 2015;107(5):djv036
4. Referans4.Ghoncheh M, Pournamdar Z, Salehiniya H. Incidence and Mortality and Epidemiology of Breast Cancer in the World. Asian Pac J Cancer Prev. 2016;17(S3):43–46.
5. Referans5.Youlden DR, Cramb SM, Yip CH, Baade PD. Incidence and mortality of female breast cancer in the Asia-Pacific region. Cancer Biol Med. 2014;11(2):101–15
6. Referans6.Polyak K. Breast cancer: origins and evolution. J Clin Invest. 2007;117(11):3155-63.
7. Referans7.Perrone, L, Tell G, Lauro RD. Calreticulin enhances the transcriptional activity of thyroid transcription factor-1 by binding to its homeodomain. J Biol Chem.1999; 274(8): 4640-45.
8. Referans8.Waser M, Mesaeli N, Spencer C, Michalak M. Regulation of calreticulin gene expression by calcium. J Cell Biol. 1997;138(3):547-57.

9. Referans9.Rooke K, Briquet-Laugier V, Xia YR, Lusis AJ, Doolittle MH. Mapping of the gene for calreticulin (Calr) to mouse chromosome 8. Mamm Genome. 1997;8(11):870-71.
10. Referans10.Smith MJ, Koch GL. Multiple zones in the sequence of calreticulin (CRP55, calregulin, HACBP), a major calcium binding ER/SR protein. EMBO J. 1989;8(12):3581-86.
11. Referans11.Coulombe, PA., Omary, MB. 'Hard' and 'soft' principles defining the structure, function and regulation of keratin intermediate filaments. Curr Opin Cell Biol.2002; 14 (1):110-22
12. Referans12.Daneau G, Boidot R, Martinive P, Feron, O Clin. Cancer Res Identification of cyclooxygenase-2 as a major actor of the transcriptomic adaptation of endothelial and tumor cells to cyclic hypoxia: effect on angiogenesis and metastases. Clin Cancer Res.2010; 16(2):410-9
13. Referans13.Ogino S, Kirkner GJ, Nosho K, Irahara N, Kure S, Shima K, et al. Cyclooxygenase-2 expression is an independent predictor of poor prognosis in colon cancer. Clin Cancer Res. 2008;14(24):8221-27.
14. Referans14.Murray, CJL., Lopez AD. Mortality by cause for eight regions of the world: Global Burden of Disease Study. Lancet.1997; 349: 1269-76
15. Referans15.Michalak M, Corbett EF, Mesaeli N, Nakamura K, Opas M. Calreticulin: one protein, one gene, many functions. Biochem J. 1999; 344:281–92
16. Referans16.Harada K, Takenawa T, Ferdous T, Kuramitsu Y, Ueyama Y. Calreticulin is a novel independent prognostic factor for oral squamous cell carcinoma. Oncol Lett. 2017;13(6):4857-62.
17. Referans17.Lwin ZM, Guo C, Salim A, Yip GW, Chew FT, Nan J, et al. Clinicopathological significance of calreticulin in breast invasive ductal carcinoma. Mod Pathol. 2010; 23(12):1559-66.
18. Referans18.Zamanian M, Qader Hamadneh LA, Veerakumarasivam A, Abdul Rahman S, Shohaimi S, Rosli R. Calreticulin mediates an invasive breast cancer phenotype through the transcriptional dysregulation of p53 and MAPK pathways. Cancer Cell Int. 2016;16:56.
19. Referans19.Coulombe PA, Wong P. Cytoplasmic intermediate filaments revealed as dynamic and multipurpose scaffolds. Nat Cell Biol. 2004;6(8):699-706.
20. Referans20.Hendrix MJ, Seftor EA, Chu YW, Trevor KT, Seftor RE. Role of intermediate filaments in migration, invasion and metastasis. Cancer Metastasis Rev. 1996;15(4):507-25.
21. Referans21.Kabir NN, , Rönnstrand L, Kazi JU. "Keratin 19 expression correlates with poor prognosis in breast cancer". Mol Biol Rep. 2014;41(12):7729-35.
22. Referans22.Solanki R, Agrawal N, Ansari M, Jain S, Jindal A. COX-2 Expression in Breast Carcinoma with Correlation to Clinicopathological Parameters. Asian Pac J Cancer Prev. 2018;19(7):1971-75.
23. Referans23.Sakoda LC, Gao YT, Chen BE, Chen J, Rosenberg PS, Rashid A, et al. Prostaglandin-endoperoxide synthase 2 (PTGS2) gene polymorphisms and risk of biliary tract cancer and gallstones: a population-based study in Shanghai, China. Carcinogenesis. 2006; 27(6):1251-56
24. Referans24.Ristimäki A, Sivula A, Lundin J, Lundin M, Salminen T, Haglund C, et al. Prognostic significance of elevated cyclooxygenase-2 expression in breast cancer. Cancer Res. 2002;62(3):632-35.
25. Referans25.Liu X, Feng C, Liu J, Liu J, Li C, Xu C, Niu et al. The importance of EGFR as a biomarker in molecular apocrine breast cancer. Hum Pathol. 2018 ;77:1-10.
26. Referans26.Shim JY, An HJ, Lee YH, Kim SK, Lee KP, Lee KS. Overexpression of cyclooxygenase-2 is associated with breast carcinoma and its poor prognostic factors. Mod Pathol. 2003;16(12):1199-204.
27. Referans27.Ghahremanfard F, Toussy JA, Kazeminezhad B, Ramezani F. Role of cyclooxygenese-2 (COX-2) expression in breast cancer differentiation and its relationship with hormone receptors status.2013; 8 (4): 235-240.