IKKɑ调控表皮角朊细胞的增殖和分化及与p63的关系  

The regulation of IKKαon the proliferation and differentiation of keratinocytes and its relationship with p63

作　　者：沈燕君[1] 辛颖[1] 周彤[2] 孙小梅[2] 姜新[2] 

机构地区：[1]吉林大学病理生物学教育部重点实验室,吉林长春130021 [2]吉林大学第一医院

出　　处：《中国实验诊断学》2016年第2期175-178,共4页Chinese Journal of Laboratory Diagnosis

基　　金：国家青年科学基金项目(81201218);吉林省科技厅国际科研合作项目(20130413032GH;20150204093SF);吉林大学白求恩B计划(2015203;2015225)

摘　　要：目的探讨IKKɑ调控表皮角朊细胞的增殖和分化及与p63的关系。方法原代分离培养野生型(wildtype,WT)和Ikkɑ基因敲除小鼠(Ikkα-/-)的表皮角朊细胞(WT-KTC及Ikkα-/-KTC),Western blot检测Ikkα-/-中p63的表达。含0.5mM Ca2+培养基刺激分化后,免疫荧光染色观察分化标记物Zonula occludens-1(ZO-1)及增殖标记物BrdU的表达。制备沉默p63表达的慢病毒并感染Ikkα-/-KTC,观察其增殖及分化能力的变化。结果在正常培养条件下,Ikkα-/-KTC中BrdU阳性细胞率显著高于WT-KTC(P<0.05)。Ikkα-/-KTC中p63高表达。高钙条件刺激分化后,WT-KTC中BrdU表达显著降低,表达ZO-1;而Ikkα-/-KTC的BrdU表达无明显变化;且无ZO-1的表达。将表达shRNA p63的慢病毒感染Ikkα-/-KTC沉默p63后,含0.5mM Ca2+培养基刺激细胞,Ikkα-/-KTC中BrdU的表达显著降低,不表达分化标记物ZO-1。结论 IKKɑ调控表皮角朊细胞的增殖和分化,且其对增殖的调控通过p63信号通路。Objective The purpose of this study is to discuss the regulation of IKKa on the proliferation and differ entiation of keratinocytes and its relationship with p63 pathway. Methods The keratinocytes was separated and cul- tured from wild-type （WT-KTC） and Ikka knockout mice （Ikk-/- KTC）,and p63 expression was detected by Western blot in those cells. WT-KTC and Ikka-/-- KTC were stimulated with the medium including high calcium （0. 5 mM）. It was observed and compared between WT-KTC and Ikka-/- KTC that the expression of Zonula occludens-1 （ZO-1） and BrdU by immunofluorescence staining,as well as p63 expression by Western blot. The proliferation and differentiation of Ikka-/- KTC under the high calcium condition were evaluated after infected with the lentivirus expressed the p63 specific shRNA. Results Under the normal cultured condition, the BrdU positive ratio of Ikka-/- KTC and p63 expression were significantly higher than that of WT-KTC. With the high calcium stimulation,the BrdU positive ratio was decreased and ZO-1 expression was detected in WT KTC, however, there was no ZO-1 expression and no change of the BrdU positive ratio in Ikka-/- KTC. When the Ikka/ KTC were infected with the lentivirus expressed p63 specific shRNA and stimulated with high calcium, the BrdU expression was inhibited, while there was still no ZO-1 expression. Conclusion IKKa regulates the proliferation and differentiation of keratinocytes,and the effect on proliferation is through p63 pathway.

关 键 词：IKKɑ 角朊细胞 增殖 分化 P63 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]