[Antisense to cyclin D1 inhibits the proliferation of glomerular mesangial cells].
OBJECTIVE
To investigate the role of Cyclin D1 in the proliferation of glomerular mesangial cells.


METHODS
We first detected the expression of Cyclin D1 protein in mesangial cells by using immunohistochemical method, set up a retroviral vector mediated genetransfer system expressing Cyclin D1 antisense RNA with recombinating techniques, and assay the proliferating situation of genetransferred mesangial cells by MTT method and flow cytometry respectively.


RESULTS
Cyclin D1 protein was expressed in the nuclei of cultured mesangial cells in vitro. The exgoenous gene was transducted into mesangial cells mediated by retrovirus and transcripted and expressed antisense RNA in high effiency. The response of antisense Cyclin D1 genetransferred mesangial cells to Endothelin-1 (OD570)(18 hr 0.105 +/- 0.009 vs 6 hr 0.096 +/- 0.013) was not significant (P > 0.05), but normal cells (18 hr 0.259 +/- 0.080 vs 6 hr 0.100 +/- 0.007) and only vector transferred cells (18 hr 0.278 +/- 0.020 vs 6 hr 0.097 +/- 0.008) were significant (P < 0.01). The progression of cell cycle was prevented from G1 to S phase by antisense Cyclin D1, and the percentage of cells of S phase in antisense Cyclin D1 genetransferred cells (12.70 +/- 3.39) was significantly reduced (P < 0.01) as compared with normal cells (29.98 +/- 3.26).


CONCLUSIONS
Cyclin D1 is a key regulatory protein of G1 progressing of cell cycle of mesangial cells, and the cellular proliferation stimulated by Endothelin-1 can be suppressed prominently in antisense Cyclin D1 gene transducted mesangial cells.
Institute of Nephrology, Beijing Medical University.
H
H
Huang
H
H
Wang
Y
Y
Lu
D000818:Animals; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D019938:Cyclin D1 / Q000494:pharmacology*; D003864:Depression, Chemical; D005920:Glomerular Mesangium / Q000166:cytology / Q000378:metabolism*; D020320:Oligoribonucleotides, Antisense; D051381:Rats
D020320:Oligoribonucleotides, Antisense; D019938:Cyclin D1