Kitamura M, Shirasawa T, Maruyama N
Department of Molecular Pathology, Tokyo Metropolitan Institute of Gerontology, Japan.
Kidney Int. 1994 Jun;45(6):1580-6. doi: 10.1038/ki.1994.208.
The aim of the present study is to clarify whether the cellular expression of a matrix-degrading metalloproteinase, transin, alters the behavior of cultured mesangial cells (MCs). The cDNA encoding rat transin was introduced into rat MCs and transcribed under the control of a Rous sarcoma virus promoter. The resulting transfectants were then investigated for cell shape, migration, proliferation, and expression of genes associated with matrix metabolism. Northern blot analysis routinely detected the transin transcript in two separate transfectants, MeTRN2 and MeTRN5. Transin expression was strong in MeTRN2, moderate in MeTRN5, but absent in mock transfectants. Immunoblot analysis revealed that these transin transfectants synthesized 59 and 62 kDa molecules, which correspond to transin gene products. Casein digestion assay detected enhanced proteolytic activity in MeTRN2 and MeTRN5. Microscopically, the transfected cells were somewhat elongated with accentuated margins compared with mock transfectants. [3H]-thymidine uptake studies revealed accelerated growth of the transfectants on a plastic substratum as well as within gel matrix. The migration of the transfectants into gel matrix was also significantly enhanced compared with that of mock transfectants. No obvious alteration, however, was found in transcripts of procollagen alpha 1(IV), laminin B2, or the metalloproteinase inhibitor TIMP. We hypothesize that the metalloproteinase transin has a potential for affecting the behavior of MCs and contributing to the pathogenesis of glomerular injury.
