Investigation of Doublecortin and Calcium/Calmodulin-dependent Protein Kinase-like-1-expressing Cells in the Mouse Colon with Acute and Chronic Mucosal Injury

WEN Quan, Mahesh MAHASETH, ZHOU Li-ping

Abstract

To identify the expression of Doublecortin and calcium/calmodulin-dependent protein kinase-like-1-expressing cells (DCAMKL-1-expressing cells) in the colon epithelium and to analyze their deviation in dextran sulfate sodium (DSS) induced colitis and colitis associated cancer. Methods A total of 60 healthy female C57 BL/6J mice were used, 40 for the DSS induced colitis model group and 20 for colitis associated cancer model group. In the former group, mice were sacrificed at day 7 after DSS administration （B group）, 3 days （C group） and 7 days （D group） after DSS withdraw, separately. The control （A group） mice were sacrificed at day 7. In the latter group, mice were gave three repetitive oral administrations of DSS and regular water after injected intraperitoneally with azoxymethane (AOM). The control group mice were injected intraperitoneally with physiological saline, and then regular water was given. All the mice were sacrificed 61 days later. DCAMKL-1 expressions were detected by both immunohistochemistry and Western blot. Results There were some DCAMKL-1-expressing cells in the normal mouse colon epithelium. Most of them were located in the base of the crypt. All DCAMKL-1-expressing cells expressed Musashi-1. In the DSS-induced colitis, the number of DCAMKL-1-expressing cells decreased 7 days after DSS administration and recovered 3 days later. The expression of DCAMKL-1 increased apparently in the mice with colitis-induced cancer. Except the base of the crypt, some of them were located in the middle portion of the crypt and some exhibited cytoplasm β-Catenin staining. Conclusion DCAMKL-1 is a putative intestinal stem cell marker. Using DCAMKL-1 as a marker for colon stem cells, we could describe the expression pattern of colon stem cells during acute and chronic mucosal injury.

 

Keywords:  Doublecortin and calcium/calmodulin-dependent protein kinase-like-1, Stem cell DSS colitis, Colitis-associated colorectal cancer 


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References


Radtke F. Clevers H. Self-renewal and cancer of the gut: two sides of a coin. Science.2005;307(5717): 1904-1909.

Qiu JM. Roberts SA. Potten CS. Cell migration in the small and large bowel shows a strong circadian rhythm. Epithelial Cell Biol, 1994 ;3(4); 137-148.

Coussens LM. Werb Z. Inflammation and cancer. Nature, 2002;420(6917):860-867.

Lin PT, Gleeson JO, Corbo JC, el al. DCAMKL1 encodes a protein kinase with homology to doublecortin that regulates microtubule polymerization. J Neurosci, 2000; 20 ( 24 ); 9152- 9161.

Zhang Y, Huang XL. Investigation of doublecortin and calcium/calmodulindependent protein kinase-like-1-expressing cells in the mouse stomach. J Gastro Hepatol, 2010; 25 ( 3): 576-582.

Cooper HS, Murthy SN, Shah RS, el al. Clinicopathologic study of dextran sulfate sodium experimental murine colitis. Lab Invest,1993;69(2):238-249.

Greten F, Eckmann L, Greten T, et al. IKKbeta links inflammation and tumorigenesis in a mouse model of colitis- associated cancer. Cell,2004; 118(3);285-296.

Cai WB, Roberts SA, Potten CS. The number of clonogenic cells in crypts in three regions of murine large intestine. Int J Radiat Biol ,1997;71 (5) s 573-579.

Potten CS, Booth C, Pritchard DM. The intestinal epithelial stem cell; the mucosal governor. Int J Exp Pathol, 1997; 78 (4):219-243.

Nishimura S, Wakabayashi N, Toyoda K, el al. Expression of Musashi-1 in human normal colon crypt cells; a possible stem cell marker of human colon epithelium. Dig Dis Sci,2003;48 (8):1523-1529.

Batlle E. Henderson JT, Beghtel H, et al. Beta-catenin and TCF mediate cell positioning in the intestinal epithelium by controlling the expression of EphB/ephrinB. Cell, 2002; 111 (2):251-263.

Potten CS, Booth C, Tudor GL, et al. Identification of a putative intestinal stem cell and early lineage marker; musashi- 1. Differentiation, 2003; 71 (1); 28-41.

Kayahara T. Sawada M. Takaishi S. elal. Candidate markers for stem and early progenitor cells. Musashi-1 and Hesl. are expressed in crypt base columnar cells of mouse small intestine. FEBS Lett. 2003; 535 (1-3): 131-135.

Allavena P. Garlanda С, Borrello MG. el at. Pathways connecting inflammation and cancer. Curr Opin Genet Dev. 2008;18(1) :3-10.

Itzkowitz SH. Yio X. Inflammation and cancer IV. Colorectal cancer in inflammatory bowel disease: the role of inflammation. Am J Physiol Gastrointest Liver Physiol. 2004 ; 287(1 ):G7-G17.

Krok KL. Lichtenstein GR. Colorectal cancer in inflammatory bowel disease. Curr Opin Gastroenterol.2004;20( 1) :43-48.

Dalerba P. Cho RW. Clarke MF. Cancer stem cells: models and concepts. Annu Rev Med.2007;58:267-284.

Carmen AB. Meirong X. Gayle AO. elal. (3-Catenin mutation in rat colon tumors initiated by 1. 2-dimethylhydrazine and 2- amino-3-methylimidazo [4. 5-f] quinoline, and the effect of post-initiation treatment with chlorophyllin and indole-3- carbinol. Carcinogenesis.2001;22(2):315-320.

Yamada Y. Oyama T. Hirose Y. elal. (3-Catenin mutation is selected during malignant transformation in colon carcinogenesis. Carcinogenesis.2003;24(1):91-97.

Miller JR. Moon RT. Signal transduction through (3-catenin and specification of cell fate during embryogenesis. Genes Dev. 1996;10(20):2527-2539.


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