Gene transfection using lipid mediated TGFβ1 sense and antisense gene expression vectors and its effects on TGFβ1 and procollagen I mRNA expression in~(60) Co-irradiated human embryo lung fibroblasts

Objective To investigate the effects on gene expression of 60 Co 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.