腺病毒载体介导的PML生长抑制因子转染人角质形成细胞的实验研究

目的探讨腺病毒介导的PML基因对角质形成细胞(KC)的生长抑制作用。方法用原位杂交和免疫组织化学法分别检测PML基因重组腺病毒的转导效率,观察培养人KC中PMLmRNA和蛋白的表达量及表达时间,MTT法检测PML基因转染后人KC的生长状况,观察转染PML基因后KC的增殖能力的变化。结果腺病毒滴度为100MO I时被转染的人KC可达100%,Ad-PML基因转染的人KC能高效表达PMLmRNA和PML蛋白,表达时间可持续2周以上,PML基因对人KC体外生长具有明显抑制作用。结论腺病毒介导的PML基因能在人KC中高效稳定地表达,能显著抑制人KC的体外生长,PML生长抑制因子有可能用于银屑病等皮肤病的基因治疗。

Experimental Study on the Keratinocytes Transfected by Adenovirus - mediated PML Growth Suppressor

Objective To investigate the inhibitory effects of adenovirus-mediated PML growth suppressor on growth of human keratinocytes.Methods Transduction efficiency of PML gene was tested by in situ hybridization(ISH).PML mRNA and PML protein expression after Ad-PML transduction was examined by means of ISH and immunohistochemistry.Human keratinocytes(KC) were placed in 24-well tissue culture plates,and transfection was carried out with Ad-PML or Ad-control.The material was analyzed by MTT assay.Results The maximal transduction efficiency of PML was obtained at a concentration of 100MOI.The Ad-PML-infected cells expressed a high level of PML mRNA and protein.A detectable level of PML was found on day 15 after viral infection.The growth rate of the Ad-PML-infected keratinocytes was significantly inhibited.Conclusion PML gene is highly and stably expressed in Ad-PML-infected keratinocytes.PML gene is able to suppress growth of human keratinocytes in vitro.Ad-PML may be a potential candidate for gene therapy of psoriasis and other dermatoses.