Farklı Hücre Hatlarında Hipoksik Koşullarda ADAMTS-2 İfadesinin Değişimi

Year 2019, Volume: 19 Issue: 1, 22 - 33, 28.05.2019

Sümeyye Aydogan Türkoğlu , Sinem Gültekin Tosun Feray Köçkar

https://doi.org/10.35414/akufemubid.428822

Cited By: 1

Abstract

ADAMTS’ler (A Disintegrin And Metalloproteaz With
Trombospondin Motifs; Trombospondin Motifli Matriks Metalloproteazlar)
dokuların yeniden düzenlenmesi, matriks yapımı ve yıkımı gibi fizyolojik
olaylarda rol aldığı gibi kanser, kas-iskelet sistemi hastalıkları,
enflamasyon, fibrozis gibi patolojik olaylarda da yer alır. ADAMTS ailesine ait
19 üye tanımlanmış ve birçok özelliğine göre gruplandırılmıştır. Bu gruplardan
ADAMTS2, 3 ve 14 kollajen ADAMTS’ leri oluşturur ve kollajen işlenmesinde görev
alır. ADAMTS2’nin önemli özelliklerinden biri kollajenlerin amino uçlarının
kesilip uzaklaştırılmasında görev alması yeni tespit edilen diğer bir
fonksiyonu  ise anti-anjiyogenik
aktiviteye sahip olmasıdır. Hipoksik
regülasyon hücresel düzeyde çok sayıda genin ifadesini etkileyen bir süreçtir.
Hücrelerde meydana gelen hipoksiya ile başta HIF-1 olmak üzere birçok gen
aktive olur. HIF-1 hücresel oksijen konsantrasyonundaki değişimleri algılayarak yanıt oluşturmadan sorumlu transkripsiyon faktörüdür.
Çalışmamızda ilk kez ADAMTS2 geninin hipoksik regülasyonu çalışılmış ve farklı
hücre hatlarında hipoksik koşullardaki değişimi mRNA seviyesinde tespit
edilmiştir. Çalışmamız
kapsamında Saos-2, MG-63 (insan kemik karsinomu), PC-3, DU-145 (insan prostat
karsinomu), MCF-7 (insan meme karsinomu), HT29, Colo-205 (insan kolon
karsinomu), MKN-45 (insan gastrik karsinomu), Panc-1, Mia PaCa-2 (insan
pankreas karsinomu), K-562 (insan kronik myeloid lösemi hücresi) ve HUVEC
(insan umblikal ven endotel hücresi) hücre hatları seçilmiş ve yapılan
analizlere göre ADAMTS2’ nin  en çok
MG-63 kemik karsinomda ifade olduğu tespit edilmiştir. Mia PaCa-2 ve K-562
hücrelerinde ise tespit edilebilir düzeyde ekspre olmadığı belirlenmiştir.
Kimyasal hipoksi modeli oluşturulmuş ve ADAMTS2 
mRNA seviyesi farklı hücre hatlarında incelenmiştir. DU-145, PC-3,
HT-29, MCF-7 ve Saos-2 hücre hatlarında mRNA düzeyinde ADAMTS2 seviyesinin
hipoksiyada arttığı tespit edilmiştir. En yüksek hipoksik cevap Saos-2 ve MCF-7
hücrelerinde gözlenmiştir.

Keywords

ADAMTS-2, Kollajen, Hipoksi, Anjiyogenez, Hücre Hattı

Variation of ADAMTS-2 Gene Expression in Hypoxic Conditions in Different Cell Lines

Abstract

ADAMTS
(A Disintegrin And Metalloproteinase with Trombospondin Motifs) is involved in
pathological events such as cancer, fibrosis, musculoskeletal disorders,
inflamation as well as physiological events such as tissue reorganization,
matrix building and destruction. 19 members of the ADAMTS family have been
identified and grouped according to many characteristics. The important
features of ADAMTS2 is processing the amino ends of the collagen and also has
an anti-angiogenic activity. Hypoxic
regulation is a process that affects the expression of many genes at the
cellular level. Hypoxia activates many genes, mainly HIF-1 transcription factor
which is responsible for detecting changes in cellular
oxygen concentration and for generating responses. For the first time, the
hypoxic regulation of ADAMTS2 gene was studied and the variation in hypoxic
conditions was determined at the mRNA level using different cell lines. We evaluated the effects of hypoxia in the
expression level of ADAMTS-2 in different cell lines namely; Saos-2, MG-63
(bone carcinoma), PC-3, DU-145 (prostate carcinoma), MCF-7 (breast carcinoma),
HT29, Colo-205 (colon carcinoma), MKN-45 (gastric carcinoma), Panc-1, Mia
PaCa-2 (pancreatic carcinoma), K-562 (chronic myeloid leukemia cell) and HUVEC
(umbilical vein endothelial cell). We found the most expression level in MG-63.
Mia PaCa-2 and K-562 cells were not expressed detectable
level of ADAMTS-2. A chemical hypoxic model was constructed and the level of
ADAMTS2 mRNA was examined. It has been
determined that ADAMTS2 mRNA level is increased in hypoxia in DU-145, PC-3,
HT-29, MCF-7 and Saos-2 cells.
The highest hypoxic response was observed in Saos-2 and MCF-7 cells.

Keywords

ADAMTS-2, Collagene, Hypoxia, Angiogenesis, Cell Line

References

• Alper, M. and Kockar, F. 2014. Il-6 Upregulates a Disintegrin and Metalloproteinase with Thrombospondin Motifs 2 (Adamts-2) in Human Osteosarcoma Cells Mediated by Jnk Pathway, Molecular and Cellular Biochemistry, 393,1,2, 165-75.
• Alper, M., Aydemir, A. T., Kockar, F., 2015. Induction of human ADAMTS-2 gene expression by IL-1 alpha is mediated by a multiple crosstalk of MEK/JNK and PI3K pathways in osteoblast like cells. GENE, 573, 2, 321-327.
• Arslan, N., 2012. Resveratrolün Hipoksi- Reoksijenasyon İle İndüklenen In Vıtro Endotel Hücresi Hasarına Etkisinin ve Nrf2’nin Olası Rolünün Araştırılması, doktora tezi, Sağlık bilimleri enstitüsü, Dokuz Eylül Üniversitesi, 202.
• Balamurugan, K., 2016. Hif-1 at the Crossroads of Hypoxia, Inflammation, and Cancer. International Journal of Cancer, 138,5, 1058-66.Bell, R.B. and White, R.P., 2000. Osteogenesis imperfecta and orthognathic surgery: case report with long- term follow-up. Int J Adult Orthodon Orthognath Surg, 15, 171-178.
• Bonaventure, J., Stanescu, R., Stanescu, V., Allain, J.C., Muriel, M.P. and Ginisty, D., 1992. Type II collagen defect in two sibs with the Goldblatt syndrome, a chondrodysplasia with dentinogenesis imperfecta, and joint laxity. Am J Med Genet, 44 ,738-53.
• Brodsky, B. and Persikov, A. V., 2005. Molecular structure of the collagen triple helix, Advances in Protein Chemistry, 70, 301-39.
• Brusselmans, K., Bono, F., Maxwell, P., Dor Y, Dewerchin, M., Collen, D., Herbert, J.M. and Carmeliet, P., 2001. Hypoxia-inducible factor 2-α (HIF-2α) is involved in the apoptotic response to hypoglycemia but not to hypoxia. Journal of Biological Chemistry, 276, 39192-39196.
• Chefetz, I., Heller, R., Galli Galli-Tsinopoulou, A., Richard, G., Wollnik, B. and Indelman, 2005. A novel homozygous missense mutation in FGF23 causes Familial Tumoral Calcinosis associated with disseminated visceral calcification. Hunt Genet, 118(2), 261-6.
• Chou, C.W. and Chen, C.C., 2008. HDAC inhibition upregulates the expression of angiostatic ADAMTS1. FEBS Letters, 582, 4059–4065.
• Colige, A., Ruggiero, F., Vandenberghe, I., Dubail, J., Kesteloot, F., Van Beeumen, J., Beschin, A., Brys, L., Lapiere, C.M., and Nusgens, B., 2005. Domains and Maturation Processes That Regulate the Activity of Adamts-2, a Metalloproteinase Cleaving the Aminopropeptide of Fibrillar Procollagens Types I-Iii and V, Journal of Biological Chemistry, 280, 41, 34397-408.
• Colige, A., Vandenberghe, I., Thiry, M., Lambert, C.A., Van Beeumen, J. and Li S.W., 2002. Cloning and characterization of ADAMTS-14, a novel ADAMTS displaying high homology with ADAMTS-2 and ADAMTS-3. J Biol Chem, 277,5756-66.
• Cömertoğlu, İ., Sarıkaya, E., Demirel, M., Akyol, S. ve Demircan, K., 2014. Role of ADAMTS Gene Family in Obstetrics and Gynecology, Derleme.
• Dubail, J., Kesteloot, F., Deroanne, C., Motte, P., Lambert, V., Rakic, J. M., Lapiere, C., Nusgens, B. and Colige, A., 2010. ADAMTS-2 functions as anti-angiogenic and anti-tumoral molecule independently of its catalytic activity. Cell Mol Life Sci, 67, 24, 4213-32.
• Dubail, J., Kesteloot, F., Deroanne, C., Motte, P., Lambert, V., Rakic, J.M., Lapiere, C., Nusgens, B., and Colige, A., 2010. Adamts-2 Functions as AntiAngiogenic and Anti-Tumoral Molecule Independently of Its Catalytic Activity, Cellular and Molecular Life Sciences, 67, 24, 4213-32.
• Forestier, I.B., Berdal, A., Vinckier, F., Ravel, T., Fryns, J.P. and Verloes, A., 2008. The genetic basis of inherited anomalies of teeth, Part 2: Syndromes with significant dental involvement. Eur J Hum Genet, 51,383-408.
• Hofer, T.P., Frankenberger, M., Mages, J., Lang, R., Meyer, P. and Hoffmann, R., 2008. Tissue-specific induction of ADAMTS2 in monocytes and macrophages by glucocorticoids. Journal of Molecular Medicine , 86, 323–32.
• Jones, G.C. and Riley, G.P., 2005. ADAMTS proteinases: a multi-domain, multi-functional family with roles in extracellular matrix turnover and arthritis. Arthritis Research and Theraphy, 7, 4, 160.
• Kantaputra, P.N., 2002. Appatently new osteodysplastic and primordial short stature with severe microdontia, opalescent tetth, and rootless molars in two. Am J Med Genet, 111, 420-8.
• Kaushal, G.P. and Shah, S.V., 2000. The new kids on the block: ADAMTSs, potentially multifunctional metalloproteinases of the ADAM family. The Journal of Clinical Investigation, 105, 10, 1335.
• Khun, K., Mayne, R. and Burgeson, R., 1987. Structure and Function of Collagen Types. Federation of American Societies for Experimental Biology, 2, 2814-2.
• Le Goff, C., Somerville, R.P., Kesteloot, F., Powell, K., Birk, D.E., Colige, A.C., and Apte, S.S., 2006. Regulation of Procollagen Amino-Propeptide Processing During Mouse Embryogenesis by Specialization of Homologous Adamts Proteases: Insights on Collagen Biosynthesis and Dermatosparaxis, Development and Disease,133,8, 1587-96.
• Livak K.J. and Schmittgen T.D., 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2 (−Delta Delta C(T)) Method. Methods 25, 4, 402–408.
• Lodish, H., Berk, A., Kaiser, C., A., Krieger, M., Scott, M., P., Bretscher, A., Ploegh, H., and Matsudaira P., 2007. Moleküler hücre biyolojisi. Ahr K., Tontonoz M., Pantages E., F., Rice E., Palme yayıncılık, W.H. Freeman and company.
• Malagelada, C., Xifro, X., Minano, A., Sabria, J. and Alvarez, J.A., 2005. Contribution of caspase-mediated apoptosis to the cell death caused by oxygen-glucose deprivation in cortical cell cultures. Neurobiology of Disease, 20, 27-37.
• Martin, E. and Shapiro, J.R., 2007. Osteogenesis imperfecta: epidemiology and pathysiology. Curr Osteoporos Rep, 5, 91-97.
• Murayama, T., Iwatsubo, R., Akiyama, S., Amano, A. and Morisaki, I., 2000. Familial hypophosphatemic vitamin D resistant rickets: dental findingts and histologic study of teeth. Oral Surg Oral Med Pathol Oral Radiol Endod, 90, 310-6.
• Ouyang, Y.B., Xu, L. and Giffard, R.G., 2005. Geldanamycin treatment reduces delayed CA I damage in mouse hippocampal organotypic cultures subjected to oxygen glucose deprivation. Neurosci Lett, 380, 229-233, (2005).
• Persikov, A.V. and. Brodsky, B., 2002. Unstable molecules form stable tissues. Proc. Nat. Acad. Sci., Piscataway, 99,1101-1103.
• Porter, S., Clark, I.M., Kevorkian, L., Edwards, D.R., 2005. The ADAMTS metalloproteinases. Biochemical Journal, 386, 15-27.
• Rankin, E.B. and Giaccia, A.J., 2016. Hypoxic Control of Metastasis, Science, 352, 175-80.
• Richard, K, Ines, D., Grant, N.,W, Dylan, R.E., 2015. The ADAMTS (A Disintegrin and Metalloproteinase with Thrombospondin motifs) family. Genom Biology, 13059, 015-0676-3
• Robert, P., M., 2011. The Extracellular Matrix: an Overview. Springer, 11-365.
• Rocks, N., Paulissen, G., El Hour, M., Quesada, F., Crahay, C., Gueders, M., Foidart, 2011. ADAM-8, a metalloproteinase, drives acute allergen induced airway inflammation. 41, 2, European Journal of Immunology, 380-391.
• Semenza, G.L., 2000. Surviving ischemia: adaptive responses mediated by hypoxia-inducible factor 1. J Clin Invest, 106,809- 812.
• Semenza, G.L., 2014. Oxygen Sensing, Hypoxia-Inducible Factors, and Disease Pathophysiology, Annu Rev Pathol, 9, 47-71.
• Semenza, G.L., Nejfelt, M.K., Chi, S.M., and Antonarakis, S.E., 1991. HypoxiaInducible Nuclear Factors Bind to an Enhancer Element Located 3' to the Human Erythropoietin Gene, Proceedingof the National Academiy of Sciences U S A, 88,13, 5680- 5684.
• Shen, N., Yan, F., Pang, J., Wu, L.C., Al-Kali, A., Litzow, M.R., and Liu, S. A., 2014. Nucleolin-Dnmt1 Regulatory Axis in Acute Myeloid Leukemogenesis. Oncotarget, 5,14, 5494-509.
• Tota, G., Coccaro, N., Zagaria, A., Anelli, L., Casieri, P., Cellamare, A., Minervini, A., Minervini, C.F., Brunetti, C., Impera, L., Carluccio, P., Cumbo, C., Specchia, G., and Albano, F., 2014. Adamts2 Gene Dysregulation in T/Myeloid Mixed Phenotype Acute Leukemia. BMC Cancer, 14, 963.
• Türkoğlu S. and Köçkar F., 2016. SP1 and USF differentially regulate ADAMTS1 gene expression under normoxic and hypoxic conditions in hepatoma cells. Gene, 575,1, 48-57.
• Wang, X., Chen, W., Zhang, J., Khan, A., Li, L., Huang, F., Qiu, Z., Wang, L., and Chen, X., 2017. Critical Role of Adamts2 (a Disintegrin and Metalloproteinase with Thrombospondin Motifs 2) in Cardiac Hypertrophy Induced by Pressure Overload, Hypertension, 69,6, 1060-1069.