Trappin-2对HaCaT细胞及银屑病跨膜模型增殖的抑制作用

Trappin-2 inhibits the proliferation of HaCaT cells and transwell psoriatic skin organ culture models

目的构建Trappin-2的真核表达体系,初步探讨Trappin-2对HaCaT细胞及银屑病跨膜模型增殖的影响。方法采用DNA重组技术构建Trappin-2的表达载体pIRES2-EGFP-Trappin-2,通过脂质体法转染人宫颈癌HeLa细胞。将Trappin-2高表达上清加入HaCaT细胞及银屑病跨膜模型中,通过3H-TdR掺入法、MTT法、流式细胞术及组化染色Ki67检测对增殖的影响。结果Trappin-2作用后MTT和cpm值变化率显示Trappin-2可抑制HaCaT细胞的代谢与DNA合成,同时可使HaCaT细胞G2+S期细胞比例下降,而G1期细胞比例增加。Trappin-2可下调银屑病皮损Ki67表达水平。结论成功将Trappin-2基因转染HeLa细胞,并进行有效表达,初步证实Trappin-2具有抑制HaCaT细胞及银屑病跨膜模型增殖的特性。

Objective To construct a eukaryotic expression vector of Trappin-2 gene and investigate its effect on the proliferation of HaCaT cells and transwell psoriatic skin organ culture models. Methods The pIRES2-EGFP-Trappin-2 expression vector was constructed by DNA recombinant technique and transfected into HeLa cells with lipofectamine^TM 2000, then the supernatant containing Trappin-2 was added to HaCaT cells and transwell psoriatic skin organ culture models. ^3H-TdR incorporation, MTT method, cell cycle analysis and expression of Ki67 were used to examine their proliferation. Results Trappin-2 significantly inhibited the prolif- eration of HaCaT cells. Trappin-2 significantly decreased the proportion of the cells in G2 ＋ S phase. Trappin-2 reduced the Ki67 expression in psoriatic skin models. Conclusion pIRES2-EGFP-Trappin-2 could be successfully transfected into HeLa cells, and the expressed Trappin-2 inhibits the proliferation of HaCaT cells and psoriatic skin models.