Araştırma Makalesi
BibTex RIS Kaynak Göster

Expression Pattern of ADAMTS-3 (A Disintegrin and Matrix Metalloproteinase Type,1 Motif 3) in Normal and Cancer Cell Lines

Yıl 2022, Cilt: 13 Sayı: 1, 40 - 47, 11.04.2022

Meltem Alper A.tuğşen Aydemir Feray Köçkar

https://doi.org/10.22312/sdusbed.1053455

Öz

ADAMTS-3 is a procollagen N- proteinase whose main function is to cleave procollagen II, the principal collagen of the cartilage and bone. This maturation process provides accurate fibril conformation for tissues. Along with collagen II, ADAMTS-3 can also process some other extracellular matrix components such as collagen III and fibronectin. ADAMTS mediated processing of the extracellular matrix components has considerable importance in both normal and pathological circumstances. Therefore, substrate specificities and the tissue expression profiles of the family members have become a hot topic. Further, altered expressions of the family members have been elucidated in some tumor types. Therefore, protumor and /or tumor suppressor functions of them is being investigated. The expression profile of the ADAMTS-3 has been studied only in a few normal and cancer tissues. Knowledge of the expression patterns of the ADAMTS-3 in cell lines are rather limited. In the present study, we determined ADAMTS-3 mRNA (sqRT-PCR) and protein expression levels (western-blot) in normal HUVEC, and cancer cell lines, namely, Saos-2, MG-63, DU-145, PC-3, Hep3B and PANC-1 cells. ADAMTS-3 was highly expressed in HUVEC, Saos-2, MG-63, and PC-3 cells. These findings will be fundamental for future studies that will be performed on ADAMTS-3 transcriptional regulation studies specific cancer types and also defining of the ADAMTS-3 contribution on the development and metastasis of specific cancer types.

Anahtar Kelimeler

ADAMTS-3 gene expression cancer cell lines sqRT-PCR western-blot

Destekleyen Kurum

TÜBİTAK

Proje Numarası

114Z025

Teşekkür

This work was financially supported by the Scientific and Technological Research Council of Turkey (TUBITAK), Project number; 114Z025.

Kaynakça

  • [1] Akyol, S., Cömertoğlu, I., Firat, R., Çakmak, Ö., Yukselten, Y., Erden, G., Ugurcu, V., Demircan, K. 2015. Effect of insulin on the mRNA expression of procollagen N-proteinases in chondrosarcoma OUMS-27 cells. Oncology Letters, 10(2),1091-1096.
  • [2] Cal, S., López-Otín, C. 2015. ADAMTS proteases and cancer. Matrix Biology, 44-46,77-85.
  • [3] Rocks, N., Paulissen, G., El Hour, M., Quesada, F., Crahay, C., Gueders, M., et al. 2008. Emerging roles of ADAM and ADAMTS metalloproteinases in cancer. Biochimie, 90,369–79.
  • [4] Wagstaff, L., Kelwick, R., Decock, J., Edwards, D.R. 2011. The roles of ADAMTS metalloproteinases in tumorigenesis and metastasis. Frontiers in Bioscience (Landmark Ed), 16,1861-72.
  • [5] Kumar, S., Rao, N., Ge, R. 2012. Emerging Roles of ADAMTSs in Angiogenesis and Cancer. Cancers, 4(4), 1252–1299.
  • [6] Porter, S., Clark, I.M., Kevorkian, L., Edwards, D.R. 2005. The ADAMTS metalloproteinases. Biochemical Journal, 386,15-27.
  • [7] Fernandes, R.J., Hirohata, S., Engle, J.M., Colige, A., Cohn, D.H., Eyre, D.R., Apte, S.S. 2001. Procollagen II amino propeptide processing by ADAMTS-3. Insights on dermatosparaxis. Journal of Biological Chemistry, 276(34), 31502-9.
  • [8] Le Goff, C., Somerville, R.P., Kesteloot, F., Powell, K., Birk, D.E., Colige, A.C., 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, 133(8), 1587-96.
  • [9] Janssen, L., Dupont, L., Bekhouche, M., Noel, A., Leduc, C., Voz, M, et al. 2016. ADAMTS3 activity is mandatory for embryonic lymphangiogenesis and regulates placental angiogenesis. Angiogenesis, 19(1), 53-65.
  • [10] Dubail, J., Apte, S.S. 2015. Insights on ADAMTS proteases and ADAMTS-like proteins from mammalian genetics. Matrix Biology,44-46, 24-37.
  • [11] Hodgkinson, L.M., Wang, L., Duncan, G., Edwards, D.R., Wormstone, I.M. 2010. ADAM and ADAMTS gene expression in native and wound healing human lens epithelial cells. Molecular Vision, 16, 2765-2776.
  • [12] Siefert, S.A, Sarkar, R. 2012. Matrix metalloproteinases in vascular physiology and disease. Vascular, 20(4), 210-216.
  • [13] Peng, W.J., Yan, J.W., Wan, Y.N, Wang, B.X., Tao, J.H., Yang, G.J., Pan, H.F., Wang, J. 2012. Matrix metalloproteinases: A review of their structure and role in systemic sclerosis. J Clin Immunol, 32(6), 1409-1414.
  • [14] Abramoff, M.D., Magalhaes, P.J., Ram, S.J. 2004. Image Processing with ImageJ. Biophotonics International, 11(7), 36-42.
  • [15] Aydemir, A.T., Alper, M., Kockar, F. 2018. SP1-mediated downregulation of ADAMTS3 gene expression in osteosarcoma models. Gene, 659,1-10
  • [16] Ernst, O., Zor, T. 2010. Linearization of the Bradford protein assay. J Vis Exp, 38, 1918.
  • [17] Timm, C., Gupta, A., Yin, J. 2015. Robust kinetics of an RNA virus: Transcription rates are set by genome levels. Biotechnology and Bioengineering, 112(8), 1655-62.
  • [18] Kaighn, M.E., Narayan, K.S., Ohnuki, Y., Lechner, J.F., Jones, L.W. 1979. Establishment and characterization of a human prostatic carcinoma cell line (PC-3) Investigative Urology, 17(1), 16-23. [19] Tai, S., Sun, Y., Squires, J.M., Zhang, H., Oh, W.K., Liang, C.Z., Huang, J. 2011. PC3 is a cell line characteristic of prostatic small cell carcinoma. The Prostate, 71(15), 1668-79.
  • [20] Park, H.J., Zhang, Y., Georgescu, S.P., Johnson, K.L., Kong, D., Galper, J.B. 2006 . Human umbilical vein endothelial cells and human dermal microvascular endothelial cells offer new insights into the relationship between lipid metabolism and angiogenesis. Stem Cell Rev.2 (2), 93-102.
  • [21] Deer, E.L, González-Hernández, J., Coursen, J.D., Shea, J.E., Ngatia, J., Scaife, C.L., Firpo, M.A., Mulvihill, S.J. 2010. Phenotype and genotype of pancreatic cancer cell lines. Pancreas. 39(4), 425-35.
  • [22] Pautke, C., Schieker, M., Tischer, T., Kolk, A., Neth, P., Mutschler, W., Milz, S. 2004. Characterization of osteosarcoma cell lines MG-63, Saos-2, and U-2 OS in comparison to human osteoblasts. Anticancer Res. 24(6),3743-8.
  • [23] Fukai, N., Apte, S.S., Olsen, B.R., Erkki, R., Engvall E. 1994. Extracellular Matrix Components. Academic Press, New York, 245, 3-28.
  • [24] Van D.R., M., Garrone, R. 1991. Collagen family of proteins. FASEB J. 5 (13), 2814-2823.
  • [25] Kevorkian, L., Young, D.A., Darrah, C., Donell, S.T., Shepstone, L., Porter, S., Brockbank, S.M., Edwards, D.R., Parker, A.E., Clark, I.M. 2004. Expression profiling of metalloproteinases and their inhibitors in cartilage. Arthritis Rheum. 50(1),131-41.
  • [26] Rocks, N., Paulissen, G., Quesada Calvo, F., Polette, M., Gueders, M., Munaut, C., Foidart, J.M., Noel, A., Birembaut, P., Cataldo, D. 2006. Expression of a disintegrin and metalloprotease (ADAM and ADAMTS) enzymes in human non-small-cell lung carcinomas (NSCLC). Br J Cancer. 94(5),724-30.
  • [27] Ogino, H., Hisanaga, A., Kohno, T., Kondo, Y., Okumura, K., Kamei, T., Sato, T., Asahara, H., Tsuiji, H., Fukata, M., Hattori, M. 2017. Secreted Metalloproteinase ADAMTS-3 Inactivates Reelin. J Neurosci. 37(12), 3181-3191.
  • [28] Porter, S., S. D. Scott, S.d., Sassoon, E.M., Williams, M.R., Jones, J.L., Girling, A.C., Ball R.Y., Edwards: D.R. 2004. Dysregulated expression of adamalysin-thrombospondin genes in human breast carcinoma. Clin Cancer Res, 10(7), 2429-40
  • [29] Zhong, S., Khalil, R.A. 2019. A Disintegrin and Metalloproteinase (ADAM) and ADAM with thrombospondin motifs (ADAMTS) family in vascular biology and disease. Biochem Pharmacol. 164,188-204.
  • [30] Pulukur,i S.M., Gondi, C.S., Lakka, S.S., Jutla, A, Estes, N., Gujrati, M., Rao, J.S. 2005. RNA interference-directed knockdown of urokinase plasminogen activator and urokinase plasminogen activator receptor inhibits prostate cancer cell invasion, survival, and tumorigenicity in vivo. J Biol Chem.280(43),36529-40.

ADAMTS-3’ ün (A Disintegrin and Matrix Metalloproteinase Type,1 Motif 3) Normal ve Kanserli Hücre Hatlarındaki Ekspresyon Paterni

Yıl 2022, Cilt: 13 Sayı: 1, 40 - 47, 11.04.2022

Meltem Alper A.tuğşen Aydemir Feray Köçkar

https://doi.org/10.22312/sdusbed.1053455

Öz

ADAMTS-3, temel görevi kıkırdak ve kemiğin temel kollajeni olan prokollajen II’ yi kesmek olan bir prokollajen amino proteinazdır. Bu olgunlaştırma işlemi, dokular için doğru fibril konformasyonunun oluşmasını sağlar. ADAMTS-3, kollajen II’ nin yanı sıra kollajen III ve fibronektin gibi diğer hücreler arası matriks bileşenlerini de işeyebilmektedir. Hücreler arası matriks bileşenlerinin ADAMTS-3 aracılı işlenmesi hem normal hem de patolojik durumlarda dikkate değer bir öneme sahiptir. Bu yüzden aile üyelerinin substrat spesifikliği ve dokulardaki ekspresyon profilleri ilgi çekici bir konu haline gelmiştir. Hatta aile üyelerinden bazılarının, bazı tümör tiplerinde ekspresyonlarının değiştiği aydınlatılmıştır. Bu yüzden protümör ve tümör supresör fonksiyonları araştırılmaktadır. ADAMTS-3’ün ekspresyon profili sadece birkaç normal ve kanserli dokuda çalışılmıştır. ADAMTS-3’ün hücre hatlarındaki ekspresyon paterni hakkındaki bilgi oldukça kısıtlıdır. Bu çalışmada normal olarak HUVEC ve kanserli hücre hatlarından, Saos-2, MG-63, DU-145, PC-3, Hep3B ve PANC-1 hücrelerinde ADAMTS-3 mRNA (sqRT-PCR) ve protein ekspresyon düzeyleri (western-blot) belirlenmiştir.HUVEC, Saos-2, MG-63 ve PC-3 hücrelerinde ADAMTS-3’ün yüksek düzeyde ifade olduğu belirlenmiştir. Bu bulgular, ADAMTS-3’ün bazı spesifik kanser türlerindeki transkripsiyonel regülasyonu ve ADAMTS-3’ün tümör gelişimine ve metastazına olan katkılarının belirlenmesi üzerine yapılacak çalışmalar için temel olacaktır.

Anahtar Kelimeler

ADAMTS-3 gen ekspresyonu kanser hücre hatları sqRT-PCR western-blot

Proje Numarası

114Z025

Kaynakça

  • [1] Akyol, S., Cömertoğlu, I., Firat, R., Çakmak, Ö., Yukselten, Y., Erden, G., Ugurcu, V., Demircan, K. 2015. Effect of insulin on the mRNA expression of procollagen N-proteinases in chondrosarcoma OUMS-27 cells. Oncology Letters, 10(2),1091-1096.
  • [2] Cal, S., López-Otín, C. 2015. ADAMTS proteases and cancer. Matrix Biology, 44-46,77-85.
  • [3] Rocks, N., Paulissen, G., El Hour, M., Quesada, F., Crahay, C., Gueders, M., et al. 2008. Emerging roles of ADAM and ADAMTS metalloproteinases in cancer. Biochimie, 90,369–79.
  • [4] Wagstaff, L., Kelwick, R., Decock, J., Edwards, D.R. 2011. The roles of ADAMTS metalloproteinases in tumorigenesis and metastasis. Frontiers in Bioscience (Landmark Ed), 16,1861-72.
  • [5] Kumar, S., Rao, N., Ge, R. 2012. Emerging Roles of ADAMTSs in Angiogenesis and Cancer. Cancers, 4(4), 1252–1299.
  • [6] Porter, S., Clark, I.M., Kevorkian, L., Edwards, D.R. 2005. The ADAMTS metalloproteinases. Biochemical Journal, 386,15-27.
  • [7] Fernandes, R.J., Hirohata, S., Engle, J.M., Colige, A., Cohn, D.H., Eyre, D.R., Apte, S.S. 2001. Procollagen II amino propeptide processing by ADAMTS-3. Insights on dermatosparaxis. Journal of Biological Chemistry, 276(34), 31502-9.
  • [8] Le Goff, C., Somerville, R.P., Kesteloot, F., Powell, K., Birk, D.E., Colige, A.C., 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, 133(8), 1587-96.
  • [9] Janssen, L., Dupont, L., Bekhouche, M., Noel, A., Leduc, C., Voz, M, et al. 2016. ADAMTS3 activity is mandatory for embryonic lymphangiogenesis and regulates placental angiogenesis. Angiogenesis, 19(1), 53-65.
  • [10] Dubail, J., Apte, S.S. 2015. Insights on ADAMTS proteases and ADAMTS-like proteins from mammalian genetics. Matrix Biology,44-46, 24-37.
  • [11] Hodgkinson, L.M., Wang, L., Duncan, G., Edwards, D.R., Wormstone, I.M. 2010. ADAM and ADAMTS gene expression in native and wound healing human lens epithelial cells. Molecular Vision, 16, 2765-2776.
  • [12] Siefert, S.A, Sarkar, R. 2012. Matrix metalloproteinases in vascular physiology and disease. Vascular, 20(4), 210-216.
  • [13] Peng, W.J., Yan, J.W., Wan, Y.N, Wang, B.X., Tao, J.H., Yang, G.J., Pan, H.F., Wang, J. 2012. Matrix metalloproteinases: A review of their structure and role in systemic sclerosis. J Clin Immunol, 32(6), 1409-1414.
  • [14] Abramoff, M.D., Magalhaes, P.J., Ram, S.J. 2004. Image Processing with ImageJ. Biophotonics International, 11(7), 36-42.
  • [15] Aydemir, A.T., Alper, M., Kockar, F. 2018. SP1-mediated downregulation of ADAMTS3 gene expression in osteosarcoma models. Gene, 659,1-10
  • [16] Ernst, O., Zor, T. 2010. Linearization of the Bradford protein assay. J Vis Exp, 38, 1918.
  • [17] Timm, C., Gupta, A., Yin, J. 2015. Robust kinetics of an RNA virus: Transcription rates are set by genome levels. Biotechnology and Bioengineering, 112(8), 1655-62.
  • [18] Kaighn, M.E., Narayan, K.S., Ohnuki, Y., Lechner, J.F., Jones, L.W. 1979. Establishment and characterization of a human prostatic carcinoma cell line (PC-3) Investigative Urology, 17(1), 16-23. [19] Tai, S., Sun, Y., Squires, J.M., Zhang, H., Oh, W.K., Liang, C.Z., Huang, J. 2011. PC3 is a cell line characteristic of prostatic small cell carcinoma. The Prostate, 71(15), 1668-79.
  • [20] Park, H.J., Zhang, Y., Georgescu, S.P., Johnson, K.L., Kong, D., Galper, J.B. 2006 . Human umbilical vein endothelial cells and human dermal microvascular endothelial cells offer new insights into the relationship between lipid metabolism and angiogenesis. Stem Cell Rev.2 (2), 93-102.
  • [21] Deer, E.L, González-Hernández, J., Coursen, J.D., Shea, J.E., Ngatia, J., Scaife, C.L., Firpo, M.A., Mulvihill, S.J. 2010. Phenotype and genotype of pancreatic cancer cell lines. Pancreas. 39(4), 425-35.
  • [22] Pautke, C., Schieker, M., Tischer, T., Kolk, A., Neth, P., Mutschler, W., Milz, S. 2004. Characterization of osteosarcoma cell lines MG-63, Saos-2, and U-2 OS in comparison to human osteoblasts. Anticancer Res. 24(6),3743-8.
  • [23] Fukai, N., Apte, S.S., Olsen, B.R., Erkki, R., Engvall E. 1994. Extracellular Matrix Components. Academic Press, New York, 245, 3-28.
  • [24] Van D.R., M., Garrone, R. 1991. Collagen family of proteins. FASEB J. 5 (13), 2814-2823.
  • [25] Kevorkian, L., Young, D.A., Darrah, C., Donell, S.T., Shepstone, L., Porter, S., Brockbank, S.M., Edwards, D.R., Parker, A.E., Clark, I.M. 2004. Expression profiling of metalloproteinases and their inhibitors in cartilage. Arthritis Rheum. 50(1),131-41.
  • [26] Rocks, N., Paulissen, G., Quesada Calvo, F., Polette, M., Gueders, M., Munaut, C., Foidart, J.M., Noel, A., Birembaut, P., Cataldo, D. 2006. Expression of a disintegrin and metalloprotease (ADAM and ADAMTS) enzymes in human non-small-cell lung carcinomas (NSCLC). Br J Cancer. 94(5),724-30.
  • [27] Ogino, H., Hisanaga, A., Kohno, T., Kondo, Y., Okumura, K., Kamei, T., Sato, T., Asahara, H., Tsuiji, H., Fukata, M., Hattori, M. 2017. Secreted Metalloproteinase ADAMTS-3 Inactivates Reelin. J Neurosci. 37(12), 3181-3191.
  • [28] Porter, S., S. D. Scott, S.d., Sassoon, E.M., Williams, M.R., Jones, J.L., Girling, A.C., Ball R.Y., Edwards: D.R. 2004. Dysregulated expression of adamalysin-thrombospondin genes in human breast carcinoma. Clin Cancer Res, 10(7), 2429-40
  • [29] Zhong, S., Khalil, R.A. 2019. A Disintegrin and Metalloproteinase (ADAM) and ADAM with thrombospondin motifs (ADAMTS) family in vascular biology and disease. Biochem Pharmacol. 164,188-204.
  • [30] Pulukur,i S.M., Gondi, C.S., Lakka, S.S., Jutla, A, Estes, N., Gujrati, M., Rao, J.S. 2005. RNA interference-directed knockdown of urokinase plasminogen activator and urokinase plasminogen activator receptor inhibits prostate cancer cell invasion, survival, and tumorigenicity in vivo. J Biol Chem.280(43),36529-40.
Toplam 29 adet kaynakça vardır.

Ayrıntılar

Birincil Dil İngilizce
Konular Sağlık Kurumları Yönetimi
Bölüm Araştırma Makaleleri
Yazarlar

Meltem Alper 0000-0001-6359-9979

A.tuğşen Aydemir Bu kişi benim 0000-0003-2803-4782

Feray Köçkar 0000-0003-2572-8391

Proje Numarası 114Z025
Yayımlanma Tarihi 11 Nisan 2022
Gönderilme Tarihi 4 Ocak 2022
Yayımlandığı Sayı Yıl 2022 Cilt: 13 Sayı: 1

Kaynak Göster

Vancouver Alper M, Aydemir A, Köçkar F. Expression Pattern of ADAMTS-3 (A Disintegrin and Matrix Metalloproteinase Type,1 Motif 3) in Normal and Cancer Cell Lines. Süleyman Demirel Üniversitesi Sağlık Bilimleri Dergisi. 2022;13(1):40-7.
 Kapak Resmi İndir

SDÜ Sağlık Bilimleri Dergisi, makalenin gönderilmesi ve yayınlanması dahil olmak üzere hiçbir aşamada herhangi bir ücret talep etmemektedir. Dergimiz, bilimsel araştırmaları okuyucuya ücretsiz sunmanın bilginin küresel paylaşımını artıracağı ilkesini benimseyerek, içeriğine anında açık erişim sağlamaktadır.