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

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.