LIM矿化蛋白-1与LIM矿化蛋白-3共转染骨髓间充质干细胞的基因表达

目的探讨LIM矿化蛋白（LIMmineralizationprotein，LMP）-1和LMP-3双基因共转染骨髓间充质干细胞（bonemesenchymalstemcells，BMSC）的表达情况。方法采用人工设计合成人LMP-1和LMP-3基因片段，分别与质粒pEGFP-N2连接，经酶切、测序鉴定后。分离培养新西兰兔BMSC，用脂质体包裹转染BMSC，按转染情况分为5组：未转染组（A组）、转染空载体组（B组）、转染LMP-1基因组（C组）、转染LMP-3基因组（D组）、LMP-1与LMP-3双基因共转染组（E组）。采用实时聚合酶链反应（real-timepolymerasechainreaction，RT-PcR）和蛋白质印迹法检测LMP-1和LMP-3的表达。结果酶切及测序表明真核表达质粒pEGFP-N2-LMP-1和pEGFP-N2-LMP-3构建成功。E组可同时较高水平表达LMP-1和LMP-3分子。对RT-PcR及蛋白质印迹法检测结果行灰度值测量并行统计学分析显示：LMP-1mRNA及蛋白水平的表达，5组间差异有统计学意义（P〈O．05），但E组与C组的差异无统计学意义（P〉O．05）；LMP-3mRNA及蛋白水平的表达，5组间差异有统计学意义（P〈O．05），且E组与D组差异也有统计学意义（P〈O．05）。结论双基因共转染的BMSC能在体外同时表达LMP-1与LMP-3，为基因修复骨缺损带来新思路。

Cotransfection and Expression of LIM mineralization protein-1 and-3 in Mesenchymal Stem Cells in vitro

Objective To study the expression of LIM mineralization protein(LMP)-1 and LMP-3 genes after cotransfecting them into bone mesenchymal stem cells(BMSC) of rabbit in vitro.Methods Fragments of LMP-1 gene and LMP-3 gene were gained through artificial synthesis,and were constructed respectively into the plasmid vector pEGFP-N2.The inserted target genes in plasmid were verified by nucleotide sequencing and enzymes.The plasmids carrying LMP-1 and LMP-3 genes were cotransfected into chondrocytes by liposome metho...