目的探讨牙齿发育相关基因金属蛋白酶解离素28(a disintegrin and metalloproteinase 28,ADAM28)在人牙乳头细胞(human dental papilla cells,HDPC)中的定位分布及作用。方法应用基因重组技术构建真核表达质粒pcDNA3.1(+)-ADAM28,通过细胞培养、基因转染HDPC后,采用免疫荧光、RT—PCR及Western blot检测ADAM28在HDPC中的表达分布。结果成功构建并鉴定ADAM28真核质粒,瞬时转染HDPC72h后,HDPC的胞质和胞膜上均表达ADAM28蛋白,而两对照组均无表达。结论ADAM28在HDPC内能被正确地翻译、表达,可能作为一种胞质蛋白调控着牙源性间充质细胞的增殖和分化。
Objective To investigate the distribution and function of tooth development related gene-ADAM28 in human dental papilla cells (HDPC). Methods Gene recombination technique was used to construct eukaryotic expression plasmid pcDNA3.1 ( + )-ADAM28. After cell culture and gene transfection into HDPC, immunofluorescence, RT-PCR and Western blot were used to detect the expression of ADAM28 in HDPC. Results ADAM28 eukaryotic plasmid was constructed and identified successfully. After transiently transfected into HDPC for 72 hours, the cytoplasms and membranes of HDPC expressed ADAM28 protein, and no expression was found in two control groups. Conclusion ADAM28 could be correctly translated and expressed in HDPC. As a plasmosin, it might regulate proliferation and differentiation of odontogenie mesenchymal cells.