Arylsulfatase B regulates colonic epithelial cell migration by effects on MMP9 expression and RhoA activation.
Arylsulfatase B (ASB; N-acetylgalactosamine-4-sulfatase; 4-sulfatase; ARSB) is the enzyme that removes 4-sulfate groups from N-acetylgalactosamine 4-sulfate, which combines with glucuronate to form the disaccharide unit of chondroitin-4-sulfate (C4S). In this study, we report how variation in expression of ASB affected the migration of human colonic epithelial cells. In the T84 cell line, derived from lung metastasis of malignant colonic epithelial cells, the activity of ASB, as well as steroid sulfatase, arylsulfatase A, and galactose-6-sulfatase, were significantly less than in normal, primary colonic epithelial cells and in the NCM460 cell line which was derived from normal colonocytes. In the T84 cells, matrix metalloproteinase 9 (MMP9), activated RhoA, and cell migration, as well as C4S content, were significantly more than in the NCM460 cells. Silencing and overexpression of ASB had inverse effects on MMP9, activated RhoA, and cell migration, as well as the C4S content, in the NCM460 and T84 cells. When ASB expression was silenced by siRNA in the NCM460 cells, MMP9 secretion increased to over 3 times the basal level, activated RhoA increased * 85%, and cell migration increased * 52%. Following overexpression of ASB, MMP9 declined 51%, activated RhoA declined * 51%, and cell migration decreased * 37%. These findings demonstrate marked effects of ASB expression on the migratory activity of colonic epithelial cells, activated RhoA, and MMP9, and suggest a potential vital role of ASB, due to its impact on chondroitin sulfation, on determination of the invasive phenotype of colonic epithelial cells.
Bhattacharyya
Sumit
S
Department of Medicine, University of Illinois at Chicago, Chicago, IL 60612, USA.
Tobacman
Joanne K
JK
D000368:Aged; D002448:Cell Adhesion; D002465:Cell Movement; D002478:Cells, Cultured; D002809:Chondroitin Sulfates / Q000032:analysis; D003106:Colon / Q000166:cytology / Q000201:enzymology*; D006025:Glycosaminoglycans / Q000032:analysis; D006801:Humans; D007413:Intestinal Mucosa / Q000166:cytology / Q000201:enzymology*; D008297:Male; D020780:Matrix Metalloproteinase 9 / Q000235:genetics* / Q000378:metabolism; D020051:N-Acetylgalactosamine-4-Sulfatase / Q000502:physiology*; D020411:Oligonucleotide Array Sequence Analysis; D020742:rhoA GTP-Binding Protein / Q000378:metabolism*
D006025:Glycosaminoglycans; C066737:RHOA protein, human; D002809:Chondroitin Sulfates; D020051:N-Acetylgalactosamine-4-Sulfatase; D020780:Matrix Metalloproteinase 9; D020742:rhoA GTP-Binding Protein