Farklı Hücre Hatlarında KLK4 Gen İfadesinin Belirlenmesi

Yıl 2022, Cilt: 12 Sayı: 2, 207 - 215, 24.12.2022

Fatma Poyrazlı , Sümeyye Aydogan Türkoğlu , Derya Babacan , Feray Köçkar

Öz

Kallikrein releated peptidaz (KLK) gen ailesi, insan genomundaki en büyük proteaz geni kümesi tarafından kodlanmaktadır ve yüksek oranda korunmuş 15 serin proteazdan oluşur. Bu ailenin doku kallikrein ve kallikrein ile ilişkili peptidazların en çok bilinen ve biyobelirteç olarak kullanılan üyesi prostata spesifik antijen (PSA- KLK3)’dir. Biyobelirteç olarak kullanılan diğer üyeleri ise, insan glandüler kallikrein 2 (KLK2) ve KLK4 (KLK-L1, PRSS17, AI2A1, ARM1, EMSP, EMSP1, PSTS TMPRSS2 ve prostaz) dür. PSA ve diğer KLK’lar arasındaki yapısal benzerlik göz önüne alındığında, ailenin kalan üyelerinin kanser biyobelirteçleri olarak potansiyel
rolü son yıllarda çokça araştırılmaktadır. Çalışmamızda PC-3/LNCaP (insan prostat karsinomu), HEP3B (insan hepatoselüler karsinomu), HT-29/SW480 (insan kolon karsinomu), ve HUVEC (insan umblikal ven endotel hücresi) hücre hatlarında KLK4’ün ifadesi mRNA ve protein düzeyinde çalışılmıştır. Çalışılan tüm hücre hatlarında ifadesinin olduğu, yine literatürle tutarlı olarak androjen bağımlı prostat kanserlerinde ise ifadesinin yüksek olduğu tespit edilmiştir. Ayrıca farklı evrelerde olan kolon kanseri hücre hatlarında,
kanser ilerleyişi ile ilişkili olarak KLK4 ifadesinin HT-29 hücrelerinde SW480 hücrelerine kıyasla yüksek bulunmuştur. Çalışmamız sonuçlarının KLK4 regülasyonunu hedefleyen kanser çalışmaları için hücre seçiminde yönlendirici olacağı düşünülmektedir

Anahtar Kelimeler

KLK4, Prostat kanseri, Kolon kanseri, Hepatoselüler karsinom, İnsan umblikal ven endotel hücresi, Human umbilical vein endothelial cell, Hepatocellular carcinoma, Colon cancer, Prostate cancer

Destekleyen Kurum

Tübitak

Proje Numarası

118Z369

Teşekkür

Bu çalışma TÜBİTAK 118Z369 nolu 1001 projesi ile desteklenmiştir. Bu çalışma 2018-2022 YÖK 100/2000 ve 2021-2022 BİDEB 2211-A Genel Yurt İçi Doktora Burs Programında bulunan Doktora öğrencisi Fatma POYRAZLI’ nın doktora tezinin bir bölümünü kapsamaktadır.

Expression of KLK4 Gene Expression Different Cell Lines

Öz

The highly conserved Kallikrein-related peptidase (KLK) gene family, encoded by the largest set of protease genes in the human genome, consists of 15 serine proteases. Prostate-specific antigen (PSA-KLK3) is the most well-known and biomarker member of tissue kallikrein and kallikrein-related peptidases. Others are human glandular kallikrein 2 (KLK2) and KLK4 (KLK-L1, PRSS17, AI2A1, ARM1, EMSP, EMSP1, PSTS TMPRSS2 and prostasis). Given the structural similarity between PSA and other KLKs, the potential role of remaining family members as cancer biomarkers has been extensively investigated in recent years. In our study,
the expression of KLK4 in PC-3/LNCaP (human prostate carcinoma), HEP3B (human hepatocellular carcinoma), HT-29/SW480 (human colon carcinoma), and HUVEC (human umbilical vein endothelial cell) cell lines was studied at the mRNA and protein level. KLK4 mRNA and protein expression were found in all studied cell lines and its expression is high in androgen-dependent prostate cancers (LNCaP), which is also consistent with the literature. In addition, a high level of KLK4 expression was found in HT-29 cells
than SW480 cells which are different stages of colon carcinoma cell lines, and this high-level expression is correlated with cancer progression. It is thought that the results of our study will be guided in the selection of cells for cancer studies targeting the regulation of KLK4.

Anahtar Kelimeler

Human umbilical vein endothelial cell, Hepatocellular carcinoma, Colon cancer, Prostate cancer, KLK4

Proje Numarası

118Z369

Kaynakça

• Avgaris, M., Mavridis K., Scorilas A. 2012. Kallikrein-related peptidases in prostate, breast, and ovarian cancers: from pathobiology to clinical relevance. Biol. Chem., 393, 301-317, DOİ: 10.1515/hsz-2011-0260.
• Christiono, S., Anggarani, W. 2021. The effect of pregnancy milk on the expression of kallikrein related peptidase-4 (KLK-4) and collagen type 1 (Coll-1) in amelogenesis. Dentino (Jur. Ked. Gigi), 6(2), 126-130.
• Clements, J., Hooper, J., Dong, Y., Harvey, T. 2001. The expanded human kallikrein (KLK) gene family: genomic organisation, tissue-specific expression and potential functions. Biol. Chem., 382(1), DOİ: 10.1515/BC.2001.002.
• Clements, J. A., Willemsen, N. M., Myers, S. A., Dong, Y. 2004. The tissue kallikrein family of serine proteases: functional roles in human disease and potential as clinical biomarkers. Crit. Rev. Clin. Lab. Sci., 41(3), 265-312, DOİ: 10.1080/10408360490471931.
• Cox, J., Khan, Z., Jackson-Boeters, L., Armstrong, J., Darling, M. 2021. Human Tissue Kallikreins in Polymorphous Adenocarcinoma: A Polymerase Chain Reaction and Immunohistochemical Study. Head Neck Pathol., 15(1), 169-178, DOİ: 10.1007/s12105-020-01196-2.
• Diamandis, E. P., Yousef, G. M., Clements, J., Ashworth, L. K., Yoshida, S., Egelrud, T., Nelson, P.S., Shiosaka, S., Little, S., Lilja, H., Stenman, U.F., Rittenhouse, H.G., Wain, H. 2000. New nomenclature for the human tissue kallikrein gene family. Clin., Chem., 46(11), 1855-1858, DOİ: 10.1093/clinchem/46.11.1855.
• Dong, Y., Kaushal, A., Bui L., Chu S Fuller, P. J., Nicklin, J., Samaratunga H., Clements, J. A. 2001. Human Kallikrein 4 (KLK4) Is Highly Expressed in Serous Ovarian Carcinomas. Clin. Cancer Res. 7(8), 2363–2371.
• Emami N., Diamandis E.P. 2008. Utility of Kallikrein-Related Peptidases (KLKs) as Cancer Biomarkers. Clin., Chem., 54(10), 1600–1607, DOİ: 10.1373/clinchem.2008.105189.
• Fuhrman-Luck R.A., Stansfield S.H., Stephens C.R., Loessner D., Clements, J. A. 2016. Prostate Cancer-Associated Kallikrein-Related Peptidase 4 Activates Matrix Metalloproteinase-1 and Thrombospondin-1. J. Proteome Res., 15(8), 2466-2478, DOİ: 10.1021/acs.jproteome.5b01148.
• Gala de Pablo, J., Armistead, F. J., Peyman, S. A., Bonthron, D., Lones, M., Smith, S., Evans, S. D. 2018. Biochemical fingerprint of colorectal cancer cell lines using label‐free live single‐cell Raman spectroscopy. J. Raman Spectrosc., 49(8), 1323-1332, DOİ: 10.1002/jrs.5389.
• Gong, W., Liu, Y., Seidl, C., Dreyer, T., Drecoll, E., Kotzsch, M., Bronger, H., Dorn, J., Magdolen, V. 2019. Characterization of kallikrein-related peptidase 4 (KLK4) mRNA expression in tumor tissue of advanced high-grade serous ovarian cancer patients. PloS one, 14(2), e0212968, DOİ: 10.1371/journal.pone.0212968.
• Gong, W., Zhu, C., Liu, Y., Muckenhuber, A., Bronger, H., Scorilas, A., Kiechle, M., Dorn, J., Magdolen, V., Dreyer T. 2021. Elevated levels of both microRNA 378 (miR-378) and kallikrein-related peptidase 4 (KLK4) mRNA are associated with an unfavorable prognosis in triple-negative breast cancer. Am. J. Transl. Res., 13(3), 1594-1606.
• Gratio, V., Beaufort, N., Seiz, L., Maier, J., Virca, G. D., Debela, M., Grebenchtchikov N., Magdolen, V., Darmoul, D. 2010. Kallikrein-related peptidase 4: a new activator of the aberrantly expressed protease-activated receptor 1 in colon cancer cells. Am. J. Pathol., 176(3), 1452-1461, DOİ: 10.2353/ajpath.2010.090523.
• Klokk, T. I., Kilander, A., Xi, Z., Wæhre, H., Risberg, B., Danielsen, H. E., & Saatcioglu, F. 2007. Kallikrein 4 is a proliferative factor that is overexpressed in prostate cancer. Cancer Res., 67(11), 5221-5230, DOİ: 10.1158/0008-5472.CAN-06-4728.
• Komatsu, N., Takata, M., Otsuki, N., Toyama, T., Ohka, R., Takehara, K., Saijoh, K. 2003. Expression and localization of tissue kallikrein mRNAs in human epidermis and appendages. J. Invest. Dermatol., 121(3), 542-549, DOİ: 10.1046/j.1523-1747.2003.12363.x.
• Kontos C.K., Scorilas A. 2012. Kallikrein-related peptidases (KLKs): a gene family of novel cancer biomarkers. Clin. Chem. Lab. Med., 50(11), 1877–1891, DOİ: 10.1515/cclm-2012-0247.
• Korkmaz, S.K., Korkmaz G.C., Pretlow G.T., Saatcioglu F. 2004. Distinctly Different Gene Structure of KLK4/KLK-L1/Prostase/ARM1 Compared with Other Members of the Kallikrein Family: Intracellular Localization, Alternative cDNA Forms, and Regulation by Multiple Hormones. DNA Cell Biol., DOİ: 10.1089/104454901750361497.
• Kraut H., Frey E. K., Werle E. 1930. Der nachweis eines kreislauf-hormons in der pankreasdrüse. Hoppeseylers. Z. Physiol. Chem., 189, 97–106, DOİ: 10.1515/bchm2.1930.189.3-4.97.
• Kryza T., Silva L.M., Bock N., Fuhrman-Luck R. A., Stephens C.R., Gao J., Samaratunga H., Lawrence M.G.,Hooper J. D.,Dong Y.,Risbridger G. P. and Clements J.A. 2017. Kallikrein-related peptidase 4 induces cancer-associated fibroblast features in prostate-derived stromal cells. Molecular Oncology, 11, 1307–1329, DOİ: 10.1002/1878-0261.12075.
• Lai J., Dong Y., Clements J.A. 2009. KLK4 (kallikrein-related peptidase 4). Atlas Genet. Cytogenet Oncol. Haematol., 13(12), DOİ: 10.4267/2042/44638.
• Li, J., Kou, M., Cui, M., Ruan, J., Cheng, Z. 2022. NaF reduces KLK4 expression by decreasing Foxo1/Runx2 expression in LS8 cells. Archives of oral biology, 133, 105311, DOİ: 10.1016/j.archoralbio.2021.105311.
• Livak, K.J., 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, DOİ: 10.1006/meth.2001.1262.
• Lundwall, A., Ylitalo, E. B., Wikström, P., Brattsand, M. 2021. Klk4t2 is a hormonally regulated transcript from the klk4 locus. Int. J. Mol. Sci., 22(23), 13023, DOİ: 10.3390/ijms222313023.
• Lundwalla, Brattsandb M. 2008. Kallikrein-related peptidases. Cell. Mol. Life Sci., 65, 2019–2038, DOİ: 10.1007/s00018-008-8024-3.
• Majumdar S., Diamandis E.P. 1999. The promoter and the enhancer region of the KLK 3 (prostate specific antigen) gene is frequently mutated in breast tumours and in breast carcinoma cell lines. Br. J. Cancer, 79(9/10), 1594–1602.
• Matsumura, M., Bhatt, A. S., Andress, D., Clegg, N., Takayama, T. K., Craik, C. S., Nelson, P. S. 2005. Substrates of the prostate‐specific serine protease prostase/KLK4 defined by positional‐scanning peptide libraries. The Prostate, 62(1), 1-13, DOİ: 10.1002/pros.20101.
• Obiezu, C. V., Scorilas, A., Katsaros, D., Massobrio, M., Yousef, G. M., Fracchioli, S., Rigault de la Longrais, I.A., Arisio R., Diamandis, E. P. 2001. Higher human kallikrein gene 4 (KLK4) expression indicates poor prognosis of ovarian cancer patients. Clin. Cancer Res., 7(8), 2380-2386.
• Rochette, P. J., Bastien, N., Lavoie, J., Guérin, S. L., Drouin, R. 2005. SW480, a p53 double-mutant cell line retains proficiency for some p53 functions. J. Mol. Biol., 352(1), 44-57, DOİ: 10.1016/j.jmb.2005.06.033.
• 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), 1668-1679, DOİ: 10.1002/pros.21383.
• Tailor, P. D., Kodeboyina, S. K., Bai, S., Patel, N., Sharma, S., Ratnani, A., Copland, J.A., She, J.X., Sharma, A. 2018. Diagnostic and prognostic biomarker potential of kallikrein family genes in different cancer types. Oncotarget, 9(25), 17876-17888., DOİ: 10.18632/oncotarget.24947.
• Tefekli A.H. 2012. Prostat Kanserinde Yeni Belirteçler ve Phi Skoru. Turk Urol Sem, 3: 61-9, DOİ: 10.5152/tus.2012.11.
• Türkoğlu, S. A., Tosun, S. G., Köçkar, F. 2019. Farklı Hücre Hatlarında Hipoksik Koşullarda ADAMTS-2 İfadesinin Değişimi. AKÜ FEMÜBİD, 19(1), 22-33, DOİ: 10.35414/akufemubid.428822.
• Seiz, L., Kotzsch, M., Grebenchtchikov, N. I., Geurts-Moespot, A. J., Fuessel, S., Goettig, P., Gkazepis, A., Wirth, M.P., Schmitt, M., Lossnitzer, A., Sweep, F.C.G.J., Magdolen, V. 2010. Polyclonal antibodies against kallikrein-related peptidase 4 (KLK4): immunohistochemical assessment of KLK4 expression in healthy tissues and prostate cancer. Biol, Chem., DOİ: 10.1515/bc.2010.033.
• Wang, P., Magdolen, V., Seidl, C., Dorn, J., Drecoll, E., Kotzsch, M., Yang, F., Schmitt, M., Schilling, O., Rockstroh, A., Clements, J.A., Loessner, D. 2018. Kallikrein-related peptidases 4, 5, 6 and 7 regulate tumour-associated factors in serous ovarian cancer. Br. J. Cancer, 119(7), 1-9, DOİ: 10.1038/s41416-018-0260-1.
• Werle E. 1934. Zur Kenntnis des haushalts des Kallikreins. Biochem. Z., 269: 415–34.
• Xi Z., Klokk T.I., Korkmaz K., Kurys P., Elbi C., Risberg B., Danielsen H., Loda M., Saatcioglu F. 2004. Kallikrein 4 is a Predominantly Nuclear Protein and Is Overexpressed in Prostate Cancer. Cancer Res., 64, 2365–2370, DOİ: 10.1158/0008-5472.CAN-03-2025.