人前脑啡肽原基因在体细胞系细胞的表达

目的 检测人前脑啡肽原基因（ｈｐｐｅ）鼠、犬，人体细胞系细胞的表达水平，评估转脑啡肽基因细胞系用于细胞镇痛的可行性。方法 采用磷酸钙共沉淀法或脂质体包裹法，将重组质粒ｐＣＭＶｈＰＰＥ（ｐＥ）单独或与ｐＲＳＶｎｅｏ（ｐＮ）共转染导入三种哺乳动物体细胞系细胞（ＮＩＨ／３Ｔ３，ＭＤＣＫ，ＷＩＳＨ）中，用Ｇ４１８培养液筛选阳性共转染细胞，扩增培养。以放射免疫法测定细胞培养上清液中脑啡肽（ＥＮＫ）的含量。结

Expression of human preproenkephalin gene in somatic cell line cells

Objective To assay the expression of exogenic enkephalin gene in mammalian somatic cell line cells and evaluate the feasibility of the gene modified cells in cell therapy for chronic pain Methods Recombinant plasmid pCMVhPPE (pE) lonely or combined with plasmid pRSVneo (pN) was transfected into somatic cell line(NIH/3T3, MDCK or WISH) cells of three kinds of mammalian using the calcium phosphate precipitation method or lipofectamine method for gene transfer The positive cotransfected cells were selected with G418 culture medium and were subcultured The concentration of enkephalin (ENK) in cell culture medium was measured with radioimmunoassay Results ENK concentration secreted by cotransfected cells (10 20ng/10 5 cells) was markedly higher than that by the cells transfected with plasmid pE (4 6ng/10 5 cells) (P<0 01) The both instant expressions were at same level (P>0 05) The rate of transfection and expression level with both plasmids by lipofectamine method were similar to those by calcium phosphate precipitation method (0 1% and 13 20ng/10 5 cells vs 0 08% and 6 10ng/10 5 cells) (P>0 05) Conclusions The human preproenkephalin gene can express well in somatic cell line cells, and the transfected cells (10 7 10 9 cells) can secret ENK enough to produce analgesic effect