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Regulation of collagenase‐3 gene expression in osteoblastic and non‐osteoblastic cell lines
Author(s) -
Selvamurugan Nagarajan,
Brown Regina J.,
Partridge Nicola C.
Publication year - 2000
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/1097-4644(20001101)79:2<182::aid-jcb20>3.0.co;2-0
Subject(s) - collagenase , northern blot , western blot , parathyroid hormone , cell culture , osteoblast , microbiology and biotechnology , gene expression , chemistry , blot , medicine , biology , endocrinology , gene , in vitro , biochemistry , enzyme , calcium , genetics
Collagenase‐3 expression in osteoblastic (UMR 106‐01, ROS 17/2.8) and non‐osteoblastic cell lines (BC1, NIH3T3) was examined. We observed that parathyroid hormone (PTH) induces collagenase‐3 expression only in UMR cells but not in BC1 (which express collagenase‐3 constitutively) or ROS and NIH3T3 cells. Since we know from UMR cells that the AP‐1 factors and Cbfa1 are required for collagenase‐3 expression, we analyzed the expression and PTH regulation of these factors by gel shift and Northern blot analysis in all cell lines. Gel mobility shift with a [ 32 P]‐labeled collagenase‐3 AP‐1 site probe indicated the induction of c‐Fos in osteoblastic cells upon PTH treatment. While c ‐ fos was induced in UMR cells, both c ‐ fos and jun B were induced in ROS cells. Since Jun B is inhibitory of Fos and Jun in the regulation of the rat collagenase‐3 gene in UMR cells, it is likely that high levels of Jun B prevent PTH stimulation of collagenase‐3 in ROS cells. When we carried out gel shift analysis with a [ 32 P]‐labeled collagenase‐3 RD ( runt domain) site probe and Northern blot analysis with a Cbfa1 specific probe, we have observed the presence of Cbfa1 in both osteoblastic and non‐osteoblastic cell lines, but there was no change in the levels of Cbfa1 RNA or protein in these cells under either control conditions or PTH treatment. From our studies above, it is evident that the expression of collagenase‐3 and its regulation by PTH in osteoblastic and non‐osteoblastic cells may be influenced by differential temporal stimulation of the AP‐1 family members, especially c‐Fos and Jun B along with the potential for posttranslational modification(s) of Cbfa1. J. Cell. Biochem. 79:182–190, 2000. © 2000 Wiley‐Liss, Inc.