American Journal of Biomedical Research
ISSN (Print): 2328-3947 ISSN (Online): 2328-3955 Website: https://www.sciepub.com/journal/ajbr Editor-in-chief: Hari K. Koul
Open Access
Journal Browser
Go
American Journal of Biomedical Research. 2017, 5(1), 1-7
DOI: 10.12691/ajbr-5-1-1
Open AccessArticle

Expression, Down-regulation and Function of CCRL2 on Three Human Cancer Cell Lines

Nadia MatterALMhana1, , Israa Zainal2 and Nahi Y. Yaseen3

1AL-Mustansirya University, College of Engineering, Environmental Engineering Department, Baghdad-Iraq

2Kirkuk University, College of Science, Chemistry Department, Kirkuk-Iraq

3Iraqi Center for Cancer and Medical Genetics, Baghdad-Iraq

Pub. Date: January 18, 2017

Cite this paper:
Nadia MatterALMhana, Israa Zainal and Nahi Y. Yaseen. Expression, Down-regulation and Function of CCRL2 on Three Human Cancer Cell Lines. American Journal of Biomedical Research. 2017; 5(1):1-7. doi: 10.12691/ajbr-5-1-1

Abstract

C-C chemokine receptor like 2 (CCRL2) is a member of the atypical chemokine receptor family; it is a hepta helical transmembrane receptor, expression of which has been shown on almost all human hematopoietic cells. CCRL2 were previously considered to be orphan receptor and as a receptor presenting its chemo attractant ligand to functional receptors. The function and expression of CCRL2 in cancer is not understood at present. Here, we investigated the expression of CCRL2 as well as the effects on cellular proliferation resulting from their knockdown in three cancer cell lines include: human cerebral glioblastoma multi form (ANGM, at passages 75-84), human cervical cancer (HeLa, at 70 passages), and human pelvic rhabdomyosarcoma (RD, at 75 passages) cell lines. In addition, all cell lines were screened for mRNA expression of CCRL2 by reverse transcription polymerase chain reaction (RT-PCR). Cell lines with detectable expression were used for knockdown experiments; and the respective influence after transfection with small interfering RNA (siRNA) concentrations (2,3,4,5,6,7 and 8) ρmol for (24 , 48 and 72) hour were determined for both CCRL2 gene and the house keeping gene GAPDH as control. The Knockdown of CCRL2 was highly successful; the expression of CCRL2 was down-regulated by over 76.0%, 89.6% and 80.7% after transfection for 48 hour to (ANGM, HeLa and RD) cell lines respectively. The results also indicated that in the CCRL2 absence there was a significant decrease in the cell proliferation, suggesting a pro-tumoral effect of CCRL2. The potential roles of CCRL2 as a novel therapeutic target and biomarker warrant further investigations.

Keywords:
chemokine receptor CCRL2 gene expression gene silencing human cancer cell lines

Creative CommonsThis work is licensed under a Creative Commons Attribution 4.0 International License. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

References:

[1]  Jason W. Griffith, Caroline L. Sokol, and Andrew D. Luster. (2014). Chemokines and Chemokine Receptors: Positioning Cells for Host Defense and Immunity. Annual Review of Immunology. 32: 659-702.
 
[2]  Bajetto A, Barbero S, Bonavia R, et al. (2001). Stromal cell-derived factor-1alpha induces astrocyte proliferation through the activation of extracellular signal-regulated kinases 1/2 pathway. J Neurochem.; 77: 1226-36.
 
[3]  Hippe A, Homey B, Mueller-Homey A. (2010). Chemokines. Recent Results Cancer Res.; 180: 35-50.
 
[4]  Frederick MJ, Henderson Y, Xu X, et al. (2000). In vivo expression of the novel CXC chemokine BRAK in normal and cancerous human tissue. Am J Pathol. 156: 1937-50.
 
[5]  Bagggiolini M, Dewald B, Moser B. (1997). Human chemokines: an update. Annu Rev Immunol.; 15: 675-705
 
[6]  Mantovani A. (1999). Chemokines: introduction & overview. Chem Immunol. 72: 1-6.
 
[7]  Murphy PM, Baggiolini M, Charo IF, et al. International union of pharmacology. XXII. Nomenclature for chemokine receptors. Pharmacol Rev.; (2000): 52: 145-176.
 
[8]  Rollins JB. Chemokines. Blood. (1997); 90: 909-28.
 
[9]  Fung H, Gersson S: Viral insertion site detection and analysis in cancer gene therapy. Gene Therapy of Cancer. Edited by: Lattime EC, Gerson SL. (2013), San Diego (CA): Elsevier, 35-46.
 
[10]  Broxmeyer HE, Kim CH. (1999). Regulation of hematopoiesis in a sea of chemokine family members with a plethora of redundant activities.Exp Hematol. 27: 1113-1123.
 
[11]  Stone, M.J., Chuang, S., Hou, X., Shoham, M., and Zhu, J.Z. (2009). Tyrosine Sulfation: An Increasingly Recognised Posttranslational Modification of Secreted Proteins. New Biotechnology; 25, 299-317.
 
[12]  Standiford TJ, Kunkel SL, Lukacs NW, et al. (1995). Macrophage inflammatory protein-1 alpha mediates lung leukocyte recruitment, lung capillary leak, and early mortality in murine endotoxemia. J Immunol. 155: 1515-1524.
 
[13]  Shimada, T., Matsumoto, M., Tatsumi, Y., Kanamaru, A., and Akira, S. A novel lipopolysaccharide inducible C-C chemokine receptor related gene in murine macrophages, FEBS Lett. (1998): 425, 490-494.
 
[14]  Fan, P., Kyaw, H., Su, K., Zeng, Z., Augustus, M., Carter, K. C., and Li, Y.. Cloning and characterization of a novel human chemokine receptor, Biochem Biophys Res Commun. (1998): 243, 264-268.
 
[15]  Galligan, C. L., Matsuyama, W., Matsukawa, A., Mizuta, H., Hodge, D. R., Howard, O. M., and Yoshimura, T. (2004). Up-regulated expression and activation of the orphan chemokine receptor, CCRL2, in rheumatoid arthritis, Arthritis Rheum 50, 1806-1814.
 
[16]  Del Prete, A., Bonecchi, R., Vecchi, A., Mantovani, A. & Sozzani, S. CCRL2, a fringe member of the atypical chemo attractant receptor family. Eur J Immunol, (2013): 43, 1418-22.
 
[17]  Migeotte, I., Franssen, J. D., Goriely, S., Willems, F., and Parmentier, M. (2002). Distribution and regulation of expression of the putative human chemokine receptor HCR in leukocyte populations, Eur J Immunol 32, 494-501.
 
[18]  Comins C, Simpson G, Relph K, Harrington K, Melcher A, Pandha H: Reoviral therapy for cancer: strategies for improving antitumor efficacy using radio- and chemotherapy. Gene Therapy of Cancer. Edited by: Lattime EC, Gerson SL. (2013), San Diego (CA): Elsevier, 185-198. 3.
 
[19]  Fung H, Gersson S: Viral insertion site detection and analysis in cancer gene therapy. Gene Therapy of Cancer. Edited by: Lattime EC, Gerson SL. (2013), San Diego (CA): Elsevier, 35-46.
 
[20]  Gujrati M, Lu Z: Targeted systemic delivery of therapeutic siRNA. Gene Therapy of Cancer. Edited by: Lattime EC, Gerson SL. (2013), San Diego (CA): Elsevier, 47-65.
 
[21]  Freshney, R. I.. Cell line provenance. Cytotechnology, (2002): 39, 3-15.
 
[22]  Phelan, M. (1988). Basic techniques for mammalian cell tissue culture. Current Protocols in Cell Biology, 7, 15-35.
 
[23]  Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C (T)) Method. Methods. 2001; 25:402-408.
 
[24]  Johnston, S. L. G. and Siegel, C. S.. Presumptive Identification of Enteroviruses with RD, HEP-2, and RMK Cell lines. Journal of Clinical Microbiology, (1990): 28(5): 1049-1050.
 
[25]  SAS. 2012. Statistical Analysis System, User's Guide. Statistical. Version 9.1th ed. SAS. Inst. Inc. Cary. N.C. USA.
 
[26]  Muller A, Homey B, Soto H, GeN, CatronD, Buchanan ME. Involvement of chemokine receptors in breast cancer metastasis. Nature. 2001; 410:50-6.
 
[27]  Ghadjar P, Coupland SE, Na IK, Noutsias M, Letsch A, Stroux A, et al. Chemokine receptor CCR6 expression level and liver metastases in colorectal cancer. J Clin Oncol Off J Am Soc Clin Oncol. 2006; 24: 1910-6.
 
[28]  Ghadjar P, Rubie C, Aebersold DM, Keilholz U. The chemokine CCL20and its receptorCCR6 in human malignancy with focuson colorectal cancer. Int J Cancer J Int Cancer. 2009; 125: 741-5.
 
[29]  Rubie C, Frick VO, Pfeil S, Wagner M, Kollmar O, Kopp B, et al. Correlation of IL-8 with induction, progression and metastatic potential of colorectal cancer. World J Gastroenterol WJG. 2007; 13: 4996-5002.
 
[30]  Rubie C, Frick VO, Wagner M, SchuldJ, Graber S, Brittner B. ELR+ CXC chemokine expression in benign and malignant colorectal conditions. BMC Cancer. 2008; 8:178.
 
[31]  Rubie C, Frick VO, Ghadjar P, Wagner M, Justinger C, Faust SK, et al. Cxc receptor-4 mRNA silencing abrogates cxcl12-induced migration of colorectal cancer cells. J Transl Med. 2011; 9: 22.
 
[32]  Zlotnik A, Yoshie O. Chemokines: a new classification system and their role in immunity. Immunity. 2000; 12: 121-7.
 
[33]  Brouwer N, Zuurman MW, Wei T, Ransohoff RM, Boddeke HW, BiberK. Induction of glial L-CCRmRNA expression in spinalcord and brain in experimental autoimmune encephalomyelitis. Glia. 2004; 46: 84-94.
 
[34]  Zuurman MW, Heeroma J, Brouwer N, Boddeke HW, Biber K. LPS-induced expression of a novel chemokine receptor (L-CCR) in mouse glial cells in vitro and in vivo. Glia. 2003; 41:327-36.
 
[35]  Zabel BA, Nakae S, Zuniga L, Kim JY, Ohyama T, Alt C, et al. Mastcell-expressed orphanreceptorCCRL2 binds chemerinandis required for optimal induction of IgE-mediated passive cutaneous anaphylaxis. J Exp Med. 2008; 205:2207-20.
 
[36]  Oostendorp J, Hylkema MN, Luinge M, Geerlings M, Meurs H, Timens W, et al. Localization and enhanced mRNA expression of the orphan chemokine receptor L-CCR in the lung in a murine model of ovalbumin-induced airway inflammation. J Histochem Cytochem Off J Histochem Soc. 2004; 52: 401-10.
 
[37]  C.C. Schimanski, S. Schwald, N. Simiantonaki, et al.(. Effect of chemokine receptors CXCR4 and CCR7 on the metastatic behavior of human colorectal cancer, Clin. Cancer Res. (2005): 11: 1743-1750.
 
[38]  Z.X. Liang, T. Wu, H. Lou, et al. Inhibition of breast cancer metastasis by selective synthetic polypeptide against CXCR4, Cancer Res. (2004) 64: 4302- 4308.
 
[39]  Zamore, P. D.. "Ancient pathways programmed by small RNAs." Science. (2002): 296(5571): 1265-9.
 
[40]  Fengqiong Yin a,b, Zhenhua Xu c, Zifeng Wang d, Hong Yao c, Zan Shen e, Fang Yu f, Yiping Tang b,Dengli Fu b, Sheng Lin g, Gang Lu d, Hsiang-fu Kung c, Wai Sang Poon d, Yunchao Huang a,⇑, Marie Chia-mi Lin. Elevated chemokine CC-motif receptor-like 2 (CCRL2) promotes cell migration and invasion in glioblastoma. Biochemical and Biophysical Research Communications 429 (2012) 168-172.
 
[41]  Israa G. Akram & Rania Georges & Thomas Hielscher & Hassan Adwan & Martin R. Berger. The chemokines CCR1 and CCRL2 have a role in colorectal cancer liver metastasis. Tumor Biol.
 
[42]  Julie Catusse1, Marion Leick1, Mareike Groch1, David J Clark2, Maike V Buchner1, Katja Zirlik1, Meike Burger. Role of the atypical chemoattractant receptor CRAM in regulating CCL19 induced CCR7 responses in B-cell chronic lymphocytic leukemia. Molecular Cancer 2010, 9:297.
 
[43]  Sehgal, C. Keener, A.L. Boynton, et al.. CXCR-4, a chemokine receptor, is overexpressed in and required for proliferation of glioblastoma tumor cells, J. Surg. Oncol. (1998): 69:99-104.
 
[44]  M. Ehtesham, K.Y. Mapara, C.B. Stevenson, et al. CXCR4 mediates the proliferation of glioblastoma progenitor cells, Cancer Lett. (2009):274:305-312.