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[Antisense to cyclin D1 inhibits the proliferation of glomerular mesangial cells]
Objectives:
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.
Insights
Cyclin D1 is crucial for mesangial cell proliferation. Suppressing Cyclin D1 with antisense RNA inhibits cell cycle progression and reduces proliferation, particularly when stimulated by Endothelin-1.
Area of Science:
- Molecular Biology
- Cell Biology
- Renal Physiology
Context:
- Glomerular mesangial cells play a key role in kidney function and disease.
- Cell proliferation is a critical process in maintaining kidney homeostasis and in the pathogenesis of renal diseases.
- Cyclin D1 is a cell cycle regulatory protein implicated in various cellular processes.
Purpose:
- To investigate the specific role of Cyclin D1 in the proliferation of glomerular mesangial cells.
- To determine if inhibiting Cyclin D1 expression can modulate mesangial cell growth.
- To assess the effect of Cyclin D1 inhibition on the response of mesangial cells to growth factors like Endothelin-1.
Summary:
- Cyclin D1 protein expression was confirmed in the nuclei of cultured mesangial cells.
- A retroviral vector system was established to express Cyclin D1 antisense RNA in mesangial cells.
- Antisense Cyclin D1 significantly inhibited cell cycle progression from the G1 to S phase, reducing the percentage of cells in S phase.
- The proliferative response of antisense Cyclin D1-transduced cells to Endothelin-1 was significantly suppressed compared to control cells.
Impact:
- This study identifies Cyclin D1 as a key regulator of mesangial cell cycle progression.
- Inhibition of Cyclin D1 offers a potential therapeutic strategy to suppress mesangial cell proliferation in kidney diseases.
- Findings contribute to understanding the molecular mechanisms underlying renal cell growth and the development of targeted therapies.