抗角蛋白K16抗体对培养角质形成细胞分化和细胞活性的影响

Regulation Study of Anti-keratin 16 Antibodies on Differentiation and Activity of Cultured Keratinocytes

目的研究抗角蛋白K16抗体对培养角质形成细胞分化和细胞活性的影响。方法鼠抗人角蛋白K16的IgG单克隆抗体作用培养的角质形成细胞,分别于作用12h,24h和36h,以同种型抗体作对照,采用定量PCR,Western blot的方法分别检测抗角蛋白K16抗体对角质形成细胞分化标志蛋白(nascent polypeptide-associated complex,NACA)和Involucrin mRNA表达的影响,以及对Involucrin和肌动蛋白合成启动蛋白(actin related protein,ARP2)表达的影响。结果 12h,24h和36h后NACA mRNA升高5.93-,3.35-和3.54-倍。12h和36h后Involucrin mRNA升高2.33-和2.0-倍。36h后Involucrin蛋白升高4.5-倍,而Arp2蛋白下降1.82-倍。结论抗角蛋白K16抗体能调节角质形成细胞的分化、影响细胞的活动力,角蛋白K16可能是维持银屑病角质形成细胞高分化和表皮增殖特性的重要因素。

Objective To study regulation of anti-keratin 16 antibodies on differentiation and activity of cultured keratinocytes. Methods Keratinocytes were primarily cultured in vitro with mouse antikeratin 16 monoclonal antibodies （mAbs） for 12,24 and 36 hours While some other keratinocytes cultured with isotype antibody served as nagtive control group. NACA and Involucrin mRNA were measured by quantitative RT-PCR. Levels of involucrin and ARP2 protein were measured by Western blot. Results NACA mRNA increased 5.93 - ,3.35 - and 3.54 - fold after 12,24 and 36 hours. While Involuerin mRNA increased 2.33 - and 2.0 - fold after 12 and 36 hours. Involucrin protein increased 4. 5 - fold, whereas Arp2 protein decreased 1.82 - fold, after 36 hours. Conclusion Anti-keratin 16 antibodies,regulated differentiation and activity of cultured keratinoeytes, could be the key factor which sustains high differentiation and proliferation charactoristic in psoriasis.