Yang Anqiao, Huang Yan, Zhang Yuting, Yang Kai, Wang Jiucun, Liu Qingmei
School of Life Sciences, Fudan University, Shanghai 200433, China.
Department of Dermatology, Jing'an District Central Hospital, Shanghai 200040, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2020 Nov 30;40(11):1564-1570. doi: 10.12122/j.issn.1673-4254.2020.11.05.
To investigate the expression of tumor-transforming gene-1 (PTTG1) in systemic sclerosis (SSc) and its role in fibrosis.
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 -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.
Compared with those in normal skin samples, the mRNA and protein expressions of PTTG1 increased significantly in the skin tissue of patients with SSc ( < 0.05). In cultured primary skin fibroblasts, the expression of PTTG1 mRNA was positively correlated with those of -SMA (R=0.8192, < 0.05), COL1A1 (R=0.6398, < 0.05), COL1A2 (R=0.316, < 0.05) and COL3A1 mRNAs (R=0.3727, < 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.
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.
研究肿瘤转化基因1(PTTG1)在系统性硬化症(SSc)中的表达及其在纤维化中的作用。
分别从21例SSc患者和22例健康皮肤患者中采集皮肤活检样本,采用实时荧光定量聚合酶链反应(RT-PCR)和免疫组织化学方法检测PTTG1的mRNA和蛋白表达。在培养的原代人皮肤成纤维细胞中,通过RNA干扰(siRNA)敲低PTTG1表达,采用RT-PCR检测PTTG1及纤维化相关基因α-SMA、COL1A1、COL1A2和COL3A1的mRNA表达水平;使用实时细胞增殖检测系统评估细胞增殖情况。
与正常皮肤样本相比,SSc患者皮肤组织中PTTG1的mRNA和蛋白表达显著增加(P<0.05)。在培养的原代皮肤成纤维细胞中,PTTG1 mRNA表达与α-SMA(R=0.8192,P<0.05)、COL1A1(R=0.6398,P<0.05)、COL1A2(R=0.316,P<0.05)和COL3A1 mRNA(R=0.3727,P<0.05)的表达呈正相关。干扰PTTG1表达可显著抑制细胞增殖,明显降低纤维化相关基因的表达,并下调成纤维细胞中胶原蛋白的表达。
PTTG1在SSc患者皮肤组织中高表达,敲低PTTG1可降低皮肤成纤维细胞活性,提示PTTG1与SSc纤维化密切相关。