Ese-3在Ca2+诱导的皮肤角质形成细胞分化中的分子机制研究

Differentiation is very important for cell against hyperproliferation and it is a essential event in caner cell formation.Ese-3 is specific expressed in epithelium cells.Many researches proved that Ese-3 plays a crucial role in tumorigenesis, but its fuction and regulating pathway, especially in cell differentiation, is largely unknown. The data from our preview studies provided strong evidence that Ese-3 could promote keratinocyte differentiation through the unknown pathway with Ca2+ inducing.Using ChIP on chip and Real time PCR analysis,4 potential Ese-3 target genes were picked out.In this study, we intend to identify the Ese-3 target gene in differentiated mice and human primary keratinocyte induced by Ca2+ in vitro and confirm the mechanism of Ese-3 regulating its target gene. In order to estimate the clinical application of Ese-3 and its downstream effectors, the diseases that related to skin cell differentiation will be demonstrated.Our research expect to reveal the mechanism of Ese-3 in keratinocyte differentiation and provide a new pathway for the diagnosis and treatment of the diseases that related to skin cell differentiation.

细胞分化作用可拮抗其过度增殖，是抑制细胞恶性转化的关键因素。Ese-3是上皮细胞特异性转录调控因子。研究显示Ese-3是抑癌基因，但其功能特别是在上皮细胞分化中的作用尚未知。我们利用基因敲除小鼠研究发现，在Ca2+诱导下，Ese-3可通过某些未知的通路或作用靶点促进皮肤角质形成细胞分化。经过ChIP on chip和Real time PCR分析，初步筛选出p63、Ese-1、Notch1、JAG-2等4个与分化相关的潜在Ese-3靶基因。本研究拟依托Ese-3敲除鼠模型，以小鼠和人角质形成细胞为对象，明确Ese-3在小鼠和人角质形成细胞分化过程中的作用靶点及调控机制；探讨临床因细胞分化异常导致的皮肤病变中Ese-3及其靶分子的表达及相关性，评价其临床应用价值。本研究有望揭示Ese-3在角质形成细胞分化中的分子网络和作用机制，为明确Ese-3在皮肤分化异常疾病中的诊疗价值提供新的理论依据。

细胞分化作用可拮抗其过度增殖，是抑制细胞恶性转化的关键因素。Ese-3是上皮细胞特异性转录调控因子。研究显示Ese-3是抑癌基因，但其功能特别是在上皮细胞分化中的作用尚未知。在本实验中，首先我们利用慢病毒稳转系统并通过流式技术、克隆形成、MTT等实验技术研究发现在角质形成细胞HaCaT中沉默Ese-3的表达可以显著促进细胞增殖；通过CHIP-sequence和荧光素酶报告试验研究发现，Ese-3可通过调节P15启动子的活性而影响HaCaT细胞的增殖；通过免疫细胞荧光实验证实当在TGF-β刺激下可通过P38调控Ese-3入核进而影响细胞的增殖。研究发现在Ca2+诱导下，通过CHIP-sequence和荧光素酶报告试验证实Ese-3可通过调节p63和JAG-2启动子的活性进而影响皮肤角质形成细胞分化，在角质形成细胞HaCaT中沉默Ese-3后显著抑制角质形成细胞HaCaT的分化。我们进一步收集鳞癌组织患者及正常皮肤标本，通过免疫组化技术证实Ese-3在皮肤鳞癌组织中表达显著降低。本研究拟依托Ese-3沉默的人角质形成细胞为对象，明确Ese-3在人角质形成细胞分化过程中的作用靶点及调控机制；探讨临床因细胞分化异常导致的皮肤病变中Ese-3及其靶分子的表达及相关性，评价其临床应用价值。本研究有望揭示Ese-3在角质形成细胞分化中的分子网络和作用机制，为明确Ese-3在皮肤分化异常疾病中的诊疗价值提供新的理论依据。