关键词: HSC-T6, TGF-β, 1, SMAD3, 增殖, 细胞周期

Abstract: Objective To explore the effect of TGF-β1 on cell proliferation,cell cycle regulation and collagen expression by HSC-T6 cell line. Methods The HSC-T6 cell proliferation was detected by MTT at 24 and 36 hours, respectively;The HSC- T6 cells were treated with TGF-β1（10 ng/ml） or not as control group. Flow cytometry was performed to measure the cell cycle;Fluorescent quantitative real time RT-PCR was used to quantify the mRNA levels of SMAD3,c-myc,cdk-2,cyclinE,EGF,HGF,Bcl-2,NF-κB,MMP1,MMP9,MMP14,TIMP-1,PAI-1,α-SMA,Collagen-I and Collagen-Ⅲ genes;Collagen-I,Collagen-Ⅲ,and α-SMA secreted by HSC-T6 cells were detected by ELISA. Results In comparison to HSC-T6 cells in control group,TGF-β1 decreased the cell counts in G₀-G₁ phase（24h: 57.3±8.5% vs 60.6±9.7%;36h: 53.0±2.2% vs 56.6±5.0%,both P>0.05）,while it increased the cells in S phase（24h:30.6±7.2% vs 26.4±10.1%;36h:35.2±3.7% vs 30.8±2.5%,both P>0.05）;We found that TGF-β1 treatment increased the mRNA levels of SMAD3,c-myc,cdk2,cyclin E, EGF,Bcl-2,NF-κB,TIMP1,PAI-1,α-SMA,and Collagn-I after treatment for 24 and 36 hours. Conversely,the mRNA level of HGF,MMP1,MMP9,MMP14,and collagen-III decreased at 24 hr,but increased at 36 hr. TGF-β1 promoted the secretion of collagen-I [24h:63.0±7.4ng/ml vs. 33.2±10.8ng/ml,P<0.05;36h:58.5±6.0ng/ml vs. 42.2±6.3ng/ml,P<0.05],and α-SMA[24h: 20.6±2.6ng/ml vs 4.2±0.7ng/ml,P<0.05;36h:59.7±14.6ng/ml vs 36.8±5.6ng/ml,P<0.05] by HSC-T6 cells after the treatment. Conclusion TGF-β1 can promote cell proliferation and the secretion of collagen of rat HSC-T6 cell lines.

Key words: HSC-T6 cells, TGF-β1, SMAD3, Proliferation, Cell cycle