| 钱霞,李小彦,肖明兵,江枫,钱静,姚宁华,周士忠,袁菊萍,陈不尤.转化生长因子β1 siRNA靶向干预受照人胚肺成纤维细胞胶原合成[J].中华放射医学与防护杂志,2015,35(12):886-890 |
| 转化生长因子β1 siRNA靶向干预受照人胚肺成纤维细胞胶原合成 |
| Effect of TGF-β1 siRNA on induction of collagen in HFL-1 cells after irradiation |
| 投稿时间：2015-08-25 |
| DOI：10.3760/cma.j.issn.0254-5098.2015.12.002 |
| 中文关键词: RNA干扰 TGF-β1 基因芯片 放射性肺纤维化 |
| 英文关键词:RNA interference transforming growth factor β1 Gene profile Radiation induced pulmonary fibrosis |
| 基金项目:南通市社会事业科技创新与示范(HS2013043) |
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| 中文摘要: |
| 目的 研究辐射对人胚肺成纤维细胞HFL-1的基因表达及对转化生长因子β1 (TGF-β1)的影响,探讨TGF-β1 siRNA对受照HFL-1细胞胶原合成的干预作用。方法 将HFL-1细胞分为单纯照射组和空白对照组,予6 MV X射线500 cGy单次照射,照后24 h以基因芯片法分析两组基因表达差异及TGF-β1的表达差异; 将HFL-1细胞分为单纯照射组、阴性对照siRNA组和TGF-β1 siRNA组,照射后24 h采用RT-PCR、ELISA、免疫组织化学及Western blot观察TGF-β1 siRNA对TGF-β1及Ⅰ、Ⅲ型胶原生成的干预作用;将HFL-1细胞分为地塞米松组、TGF-β1 siRNA组及TGF-β1 siRNA联合地塞米松组,以RT-PCR、免疫组织化学及Western blot观察照射后24 h不同干预方法对TGF-β1及Ⅰ、Ⅲ型胶原生成的影响。结果 芯片检测显示HFL-1细胞受照后基因表达谱明显改变,TGF-β1基因上调;单纯照射组TGF-β1基因(t=-18.580, P<0.001)、TGF-β1蛋白(t=-12.445, P<0.001)和羟脯氨酸(HYP)(t=-3.987,P=0.016)表达明显高于空白对照组。与单纯照射组比较,TGF-β1 siRNA组、地塞米松组和联合组的TGF-β1(t=13.235、6.943、-17.431,P<0.05)、α-SMA(t=12.548、6.966、13.988,P<0.05)、Ⅰ型(t=12.433、8.164、16.315,P<0.05)及Ⅲ型胶原(t=5.919、3.164、9.420,P<0.05)的表达均明显下降,且TGF-β1 siRNA组优于地塞米松组(t=7.110、7.976、4.196、5.935,P<0.05)。 结论 放射诱导HFL-1细胞基因表达谱明显改变,TGF-β1上调;TGF-β1 siRNA能有效沉默TGF-β1基因,干预Ⅰ、Ⅲ型胶原的产生;TGF-β1 siRNA干预胶原形成优于地塞米松。 |
| 英文摘要: |
| 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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