TY - JOUR
T1 - Identification of vitamin D-stimulated alkaline phosphatase in IEC-6 cells, a rat small intestine crypt cell line
AU - Jeng, Yow Jiun
AU - Watson, Cheryl S.
AU - Thomas, Mary L.
PY - 1994/6
Y1 - 1994/6
N2 - The IEC-6 cell line was derived from newborn rat small intestinal crypts and maintains characteristics of nontransformed crypt epithelial cells. We have reported that alkaline phosphatase (ALP) activity, a differentiation marker for intestinal epithelial cells, is regulated in IEC-6 cells by 1,25- dihydroxycholecalciferol (1,25(OH)2D3). Using reverse transcription polymerase chain reaction, we determined that the ALP in IEC-6 cells is the liver/bone/kidney ALP (LALP) isoenzyme and not the mature enterocyte form. Levamisole, a specific blocker for LALP, blocked the ALP activity of IEC-6 cells. Kinetic studies showed that 1,25(OH)2D3 increases V(m) values of ALP in IEC-6 cells while K(m) values remain the same. Northern analysis of LALP mRNA levels in IEC-6 cells indicated that 10-7 M 1,25(OH)2D3 acts by increasing alkaline phosphatase activity at the gene expression level.
AB - The IEC-6 cell line was derived from newborn rat small intestinal crypts and maintains characteristics of nontransformed crypt epithelial cells. We have reported that alkaline phosphatase (ALP) activity, a differentiation marker for intestinal epithelial cells, is regulated in IEC-6 cells by 1,25- dihydroxycholecalciferol (1,25(OH)2D3). Using reverse transcription polymerase chain reaction, we determined that the ALP in IEC-6 cells is the liver/bone/kidney ALP (LALP) isoenzyme and not the mature enterocyte form. Levamisole, a specific blocker for LALP, blocked the ALP activity of IEC-6 cells. Kinetic studies showed that 1,25(OH)2D3 increases V(m) values of ALP in IEC-6 cells while K(m) values remain the same. Northern analysis of LALP mRNA levels in IEC-6 cells indicated that 10-7 M 1,25(OH)2D3 acts by increasing alkaline phosphatase activity at the gene expression level.
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U2 - 10.1006/excr.1994.1152
DO - 10.1006/excr.1994.1152
M3 - Article
C2 - 8187827
AN - SCOPUS:0028361865
SN - 0014-4827
VL - 212
SP - 338
EP - 343
JO - Experimental Cell Research
JF - Experimental Cell Research
IS - 2
ER -