南方医科大学学报

南方医科大学学报 ›› 2020, Vol. 40 ›› Issue (11): 1564-1570.doi: 10.12122/j.issn.1673-4254.2020.11.05

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垂体肿瘤转化基因1在系统性硬化症中的表达及作用

杨安桥，黄 炎，张玉婷，杨 凯，王久存，刘庆梅   

  • 出版日期:2020-11-20 发布日期:2020-11-23

Expression of pituitary tumor-transforming gene-1 and its pathogenic role in systemic sclerosis

  • Online:2020-11-20 Published:2020-11-23

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摘要： 目的 研究垂体肿瘤转化基因（PTTG1）在系统性硬化症（SSc）中的表达及其对成纤维细胞的作用。方法 收集皮肤活检组织，其中SSc患者组21例，正常组22例。分别应用实时荧光定量聚合酶链式反应法（Real-time PCR）检测皮肤组织中PTTG1基因表达水平；应用免疫组化方法检测皮肤组织中PTTG1蛋白表达水平；应用小干扰RNA（siRNA）降低成纤维细胞中PTTG1基因表达，通过 Real-time PCR 方法检测细胞中 PTTG1 及与细胞纤维化密切相关的几个基因α-SMA、COL1A1、COL1A2、COL3A1的表达变化；通过细胞实时增殖检测系统检测细胞增殖。结果 与正常对照相比，SSc患者皮肤组织中PTTG1的表达水平明显升高，差异具有统计学意义（P<0.05）；与正常对照相比，SSc患者皮肤组织中阳性细胞增多，PTTG1蛋白表达明显升高；原代皮肤成纤维细胞中PTTG1表达水平和α-SMA、COL1A1、COL1A2、COL3A1均存在正相关，差异具有统计学意义（R2=0.8192，P<0.05；R2=0.6398，P<0.05；R2=0.316，P<0.05；R2=0.3723，P<0.05）；同时，原代皮肤成纤维细胞中PTTG1干扰后，细胞增殖被显著抑制，纤维化相关基因（COL1A1、COL1A2、PAI-1）表达下降，胶原蛋白的基因表达降低。结论 SSc患者中存在PTTG1过度表达，干扰PTTG1可降低成纤维细胞活性，提示PTTG1与SSc纤维化程度密切相关。

关键词: 垂体肿瘤转化基因, 系统性硬化症, 纤维化, 细胞增殖, 成纤维细胞

Abstract: Objective To investigate the expression of tumor-transforming gene-1 (PTTG1) in systemic sclerosis (SSc) and its role in fibrosis. Methods Skin biopsy samples were collected from 21 patients with SSc and 22 patients with healthy skin for detecting the mRNA and protein expressions of PTTG1 using real-time PCR (RT-PCR) and immunohistochemistry, respectively. In cultured primary human dermal fibroblasts, PTTG1 expression was knocked down via RNA interference (siRNA), and the mRNA expression levels of PTTG1 and the fibrosis-related genes α-SMA, COL1A1, COL1A2, and COL3A1 were detected using RT-PCR; the proliferation of the cells was assessed using a real-time cell proliferation detection system. Results Compared with those in normal skin samples, the mRNA and protein expressions of PTTG1 increased significantly in the skin tissue of patients with SSc (P<0.05). In cultured primary skin fibroblasts, the expression of PTTG1 mRNA was positively correlated with those of α-SMA (R2=0.8192, P<0.05), COL1A1 (R2=0.6398, P<0.05), COL1A2 (R2=0.316, P<0.05) and COL3A1 mRNAs (R2=0.3727, P<0.05). Interference of PTTG1 expression significantly inhibited the cell proliferation, obviously lowered the expressions of fibrosis-related genes, and down-regulated the expression of collagen in the fibroblasts. Conclusion PTTG1 is highly expressed in skin tissues of patients with SSc, and PTTG1 knockdown can reduce the activity of the dermal fibroblasts, suggesting a close correlation of PTTG1 with fibrosis in SSc.

Key words: tumor-transforming gene-1, systemic sclerosis, fibrosis, cell proliferation, fibroblasts