Objective      To investigate the expression of rat hyperplasia suppressor gene (rHSG) in rat glioma cell as well as the effect of rHSG on rat glioma cell proliferation

and to explore the mechanism.       Methods       The rat glioma cells (C6 cells) were transfected with recombinant adenovirus vector containing rHSG (Adv-rHSG-GFP). Western blotting and immunohistochemical assay were applied for determining the expressi

on of rHSG protein. The effect of rHSG on cell cycle was analyzed by flow cytometry. Together with MTT assay

cytometry and plate cloning formation method were used for identifying the effect of rHSG on cell proliferation. The expression of proliferating cell nuclear antigen (PCNA) and cell cycle regulatory proteins p27

Kip1

  and p21

Cip1

were determined by Western blotting.       Results       It was observed that in C6 cells transfected with Adv-rHSG-GFP

the protein expression level of rHSG was higher than that in untransfected cells and cells transfected with Adv-GFP (

P

0.01). The results from flow cytometry indicated that the cell cycle of C6 cells transfected with Adv-rHSG-GFP was arrested in G

0

/G

1

phase. MTT assay

cytometry and plate cloning formation results showed that the proliferation of C6 cells transfected with Adv-rHSG-GFP was markedly inhibited (

P

0.01). The expression levels of p27

Kip1 

and p21

Cip1

  were increased while that of PCNA was decreased in C6 cells transfected with Adv-rHSG-GFP

as compared to untransfected cells and cells transfected with Adv-GFP (

P

0.05

P

0.01).       Conclusion       The over-expression of rHSG can suppress the proliferation of rat glioma cells

with the possible mechanism of cell cycle regulation.