角蛋白K14反义寡核苷酸对角质形成细胞增殖的影响

目的：研究角蛋白K14反义寡核苷酸(ASODN)对人角质形成细胞(KC)体外增殖活性的影响。方法：利用脂质体将人工合成的正义、反义及错配K14寡核苷酸基因片段导人体外培养的角质形成细胞,应用细胞生长抑制实验、透射电镜(TEM)和流式细胞仪(FCM)检测ASODN对角质形成细胞的增生、超微结构改变和细胞增殖周期的影响。结果：脂质体介导的K14反义寡核苷酸组KC增殖活性受到明显抑制;电镜下可见KC增殖活性受到抑制的改变,角蛋白合成明显减少;流式细胞仪检测见细胞周期发生明显改变,G1期细胞百分率上升,S期细胞百分率下降。而正义寡核苷酸组、错义寡核苷酸组及空白对照组均无此改变。结论：K14反义寡核苷酸可抑制KC体外增殖活性,应用反义寡核苷酸技术封闭K14基因,有望为银屑病的基因治疗提供新的思路。

The effects of antisense oligonucleotides targeting keratin 14 on the proliferation of keratinocytes

Objective: To evaluate the inhibitory effects of antisense oligonucleotides targeting keratin 14 (K14) on the proliferation of keratinocytes. Methods: The antisense, sense and mismatched oligonucleotides with lipofectin for K14 gene were synthesized and transfected individually to keratinocytes cultured in vitro. The inhibition of proliferation was studied by MTT assay. The changes of cell cycle and cell morphology were detected by flow cytometry (FCM) and transmission electron microscopy (TEM), respectively. Results: The cell proliferation was inhibited markedly by K14 antisense oligonucleotides with lipofectin. The cell growth cycle and cell activation were inhibited effectively by antisense oligodeoxynucleotides, whereas those treated with sense and mismatched oligonucleotides were not. Conclusions: Antisense oligodeoxynucleotides conjugated with lipofectin might be a hopeful method to inhibit the proliferation of keratinocytes by inhibiting the expression of K14. Further investigations are needed for exploring a novel gene therapeutic approach for psoriasis.