| Qian Xia,Li Xiaoyan,Xiao Mingbing,Jiang Feng,Qian Jing,Yao Ninghua,Zhou Shizhong,Yuan Juping,Chen Buyou.Effect of TGF-β1 siRNA on induction of collagen in HFL-1 cells after irradiation[J].Chinese Journal of Radiological Medicine and Protection,2015,35(12):886-890 |
| Effect of TGF-β1 siRNA on induction of collagen in HFL-1 cells after irradiation |
| Received:August 25, 2015 |
| DOI：10.3760/cma.j.issn.0254-5098.2015.12.002 |
| KeyWords:RNA interference transforming growth factor β1 Gene profile Radiation induced pulmonary fibrosis |
| FundProject:南通市社会事业科技创新与示范(HS2013043) |
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| Abstract:: |
| Objective To investigate the effects of radiation on gene expressions of TGF-β1 in human fetal lung fibroblast cells (HFL-1), and to explore the effect of TGF-β1 siRNA on radiation-induced collagen. Methods HFL-1 cells were divided into irradiation and control group. At 24 h of 500 cGy of 6 MV X-rays irradiation, the cellular gene expressions including TGF-β1 were assayed by a gene chip. HFL-1 cells were divided into irradiation, negative control siRNA and TGF-β1 siRNA groups. The effects of TGF-β1 siRNA on TGF-β1 and collagen type-Ⅰ and -Ⅲ after irradiation were detected by RT-PCR, ELISA, immunohistochemistry and Western blot. In addition, HFL-1 cells were divided into irradiation, dexamethasone, TGF-β1 siRNA, and TGF-β1 siRNA combined with dexamethasone groups. After 24 h of irradiation, the expressions of TGF-β1, collagen type-Ⅰ and -Ⅲ were determined by RT-PCR, immunohistochemistry and Western blot, respectively.Results Gene expression profile of HFL-1 cells changed after exposure to radiation and TGF-β1 mRNA was up-regulated. The expression of TGF-β1 mRNA (t=-18.580, P<0.001) and protein (t=-12.445, P<0.001) in the irradiated cells were higher than those in the control group. The expression of HYP (t=-3.987,P=0.016) was consistent with that of TGF-β1. The expression of TGF-β1 (t=13.235, 6.943 and -17.431,P<0.05), α-SMA (t=12.548, 6.966,13.988,P<0.05), collagen type-I (t=12.433, 8.164,16.315,P<0.05) and collagen type-Ⅲ (t=5.919, 3.164, 9.420,P<0.05) in the groups of TGF-β1 siRNA, dexamethasone and TGF-β1 siRNA combined with dexamethasone were dramatically suppressed in comparison with those in the irradiation alone group. Moreover, the expression levels of TGF-β1 (t=7.110,P<0.05), α-SMA (t=7.976,P<0.05), collagen type-Ⅰ (t=4.196,P<0.05) and collagen-Ⅲ (t=5.935,P<0.05) were significantly decreased in the TGF-β1 siRNA group compared to the dexamethasone group. Conclusions Gene expression profile of HFL-1 cells is significantly changed with exposure to X-ray irradiation and up-regulation of TGF-β1 in HFL-1 cells. TGF-β1 siRNA effectively inhibits TGF-β1 expression and interfers the production of collagens. Moreover, the anti-fibrosis effect of TGF-β1 siRNA is better than that of dexamethasone. These results provide insights in ameliorating radiation-induced pulmonary fibrosis. |
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