| LIU Chunjie,WANG Dewen,ZHANG Zhaoshan,et al.Gene transfection using lipid mediated TGFβ1 sense and antisense gene expression vectors and its effects on TGFβ1 and procollagen I mRNA expression in 60Co-irradiated human embryo lung fibroblasts[J].Chinese Journal of Radiological Medicine and Protection,2001,21(2):103-106 |
| Gene transfection using lipid mediated TGFβ1 sense and antisense gene expression vectors and its effects on TGFβ1 and procollagen I mRNA expression in 60Co-irradiated human embryo lung fibroblasts |
| Received:June 18, 2000 |
| DOI： |
| KeyWords:Transforming growth factor beta 1 Gene transfection |
| FundProject:全军九五指令基金资助项目(96L011) |
| Author Name | Affiliation | | LIU Chunjie | Beijing Institute of Radiation Medicine, Beijing 100850, China | | WANG Dewen | Beijing Institute of Radiation Medicine, Beijing 100850, China | | ZHANG Zhaoshan | 北京生物工程研究所 | | 龙建银 | 北京基础医学研究所 | | 王会信 | 北京基础医学研究所 | | 高亚兵 | Beijing Institute of Radiation Medicine, Beijing 100850, China | | 熊呈琦 | Beijing Institute of Radiation Medicine, Beijing 100850, China | | 彭瑞云 | Beijing Institute of Radiation Medicine, Beijing 100850, China | | 崔雪梅 | Beijing Institute of Radiation Medicine, Beijing 100850, China | | 宋良文 | Beijing Institute of Radiation Medicine, Beijing 100850, China |
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| Abstract:: |
| Objective To investigate the effects on gene expression of 60Co-irradiated human embryo lung fibroblasts after gene transfection using lipid mediated TGFβ1 sense and antisense gene expression vectors. Methods TGFβ1 sense and antisense gene expression vectors were transfected using a lipid mediated method.Gene expression was analysed by RNA dot blot. Results HELFs irradiated with 5 Gy were transfected with an expression vector encoding the human TGFβ1 sense or antisense gene under control of the mouse mammary tumor virus long terminal repeat(MMTV-LTR)promotor/enhance sequence(pMAMneo-TGFβ1,or pMAMneo-anti TGFβ1).The transfected cells elected by G418 resistance were cultured in DMEM containing dexamethasone.The chromosomal DNA and RNA were extracted.Positive reaction was showed from chromosomal DNA by a PCR method of neo-specific primers and DNA dot blot with Dig-labelling neo-specific probe.RNA dot blot analysis showed that TGFβ1 mRNA level of the cells transfected with pMAM neo-anti TGFβ1 decreased,but that of transfected with pMAM neo-TGFβ1 increased.For procollagen Ⅰ mRNA,the transfected pMAM neo-anti TGFβ1 was lower than untransfected cells and the transfected pMAM neo-TGFβ1 was higher. Conclusion After TGFβ1 sense and antisense gene transfection,TGFβ1 mRNA level of the cells transfected with TGFβ1 antisense gene decreased,but that with TGFβ1 sense gene increased.For procollagen Ⅰ mRNA,the cells transfected with TGFβ1 antisense gene was lower than untransfected cells and the cells transfected with TGFβ1 sense gene was higher than untransfected cells. |
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