Liu Jin-Juan, Yang Hong-Fa, Li Yong-Jian, Chen Yan-Ming
Dermatology & STD Department, the Second Hospital University of South China, Hengyang 421001, China.
Cardiovascular Medicine Department, the Second Hospital University of South China, Hengyang 421001, China.
Sichuan Da Xue Xue Bao Yi Xue Ban. 2019 Sep;50(5):654-659.
To investigate the expression of β-catenin in the skin lesions of patients with systemic scleroderma (SSc) and its effect on epithelial-mesenchymal transition (EMT) of human epidermal keratinocytes.
The expression of β-catenin, Snail1 and E-cadherin in the skin lesions sample of 45 SSc patients and normal skin sample from 20 healthy adults was detected with SP immunohistochemistry. HaCaT, the human epidermal keratinocytes, were treated with different concentrations of Wnt10b (0 ng/mL (control), 2 ng/mL and 4 ng/mL) for 48 h. then detected the localization of β-catenin in HaCaT cells by immunofluorescence assay, determined the mRNA levels of Snail1 and Snail2 in HaCaT cells by real-time fluorescent quantitative PCR, detected the proteins expression of β-catenin, Vimentin, N-cadherin and E-cadherin in HaCaT cells by Western blot.
The positive rates of β-catenin, Snail1 and E-cadherin in skin lesions of SSc patients were 100%, 88.89% and 2.22% respectively, while in healthy adult skin, the corresponding positive rates were 0%, 10.00%, and 95.00%. The difference between the two groups was significant. Compared with control group, treatment with different concentrations of Wnt10b (2 ng/mL and 4 ng/mL) induced up-regulation of β-catenin expression and promoted translocation of β-catenin from cytoplasm to nucleus, increased the mRNA levels of Snail1 and Snail2 ( < 0.05), and up-regulated the proteins expression of Vimentin, N-cadherin, down-regulated the E-cadherin protein expression in HaCaT cells ( < 0.05).
Abnormally activated Wnt/β-catenin signaling pathway and abnormally expressed EMT-related proteins are observed in SSc lesions. Activation of Wnt/β-catenin signaling pathway may promote EMT in HaCaT cells.
探讨β-连环蛋白在系统性硬化症（SSc）患者皮肤病变中的表达及其对人表皮角质形成细胞上皮-间质转化（EMT）的影响。
采用SP免疫组织化学法检测45例SSc患者皮肤病变样本及20例健康成年人正常皮肤样本中β-连环蛋白、Snail1和E-钙黏蛋白的表达。将人表皮角质形成细胞HaCaT用不同浓度的Wnt10b（0 ng/mL（对照组）、2 ng/mL和4 ng/mL）处理48 h。然后通过免疫荧光法检测HaCaT细胞中β-连环蛋白的定位，通过实时荧光定量PCR测定HaCaT细胞中Snail1和Snail2的mRNA水平，通过蛋白质印迹法检测HaCaT细胞中β-连环蛋白、波形蛋白、N-钙黏蛋白和E-钙黏蛋白的蛋白表达。
SSc患者皮肤病变中β-连环蛋白、Snail1和E-钙黏蛋白的阳性率分别为100%、88.89%和2.22%，而在健康成年人皮肤中，相应的阳性率分别为0%、10.00%和95.00%。两组间差异有统计学意义。与对照组相比，不同浓度Wnt10b（2 ng/mL和4 ng/mL）处理可诱导β-连环蛋白表达上调，促进β-连环蛋白从细胞质向细胞核转位，增加HaCaT细胞中Snail1和Snail2的mRNA水平（<0.05），并上调波形蛋白、N-钙黏蛋白的蛋白表达，下调HaCaT细胞中E-钙黏蛋白的蛋白表达（<0.05）。
在SSc病变中观察到Wnt/β-连环蛋白信号通路异常激活及EMT相关蛋白异常表达。Wnt/β-连环蛋白信号通路的激活可能促进HaCaT细胞的EMT。
