Grzegrzolka Jedrzej, Wojtyra Patrycja, Biala Martyna, Piotrowska Aleksandra, Gomulkiewicz Agnieszka, Rys Janusz, Podhorska-Okolow Marzenna, Dziegiel Piotr
Department of Histology and Embryology, Wroclaw Medical University, Wroclaw, Poland
Department of Histology and Embryology, Wroclaw Medical University, Wroclaw, Poland.
Anticancer Res. 2017 Oct;37(10):5485-5493. doi: 10.21873/anticanres.11978.
BACKGROUND/AIM: As a result of activation of transcription factors engaged in epithelial-mesenchymal transition (EMT), such as Twist, inhibition of epithelial markers and an increased expression of mesenchymal markers are observed. One of the specific markers of cancer-associated fibroblasts is podoplanin (PDPN) - a mucin-type membrane glycoprotein. The aim of this work was to study the localisation and intensity of expression of Twist and PDPN on the mRNA and protein level in cases of invasive ductal breast carcinoma (IDC), and its association with patients' clinico-pathological data.
The study included archival material in a form of 80 paraffin IDC blocks and 11 IDC fragments frozen in liquid nitrogen. Immunohistochemical expression of Twist and PDPN was evaluated using light microscope and semiquantitative scale for evaluation of nuclear expression or immunoreactive scale (IRS) for evaluation of cytoplasmic expression. Material was isolated from frozen IDC fragments using laser micro-dissection (from cancer and stromal cells, separately) and was used to perform real-time PCR.
Twist expression was higher in stromal cells in comparison to cancer cells. Analysis of patients' survival rate showed, that higher expression of Twist in cancer cells was associated with shorter overall survival time and shorter event-free survival time. The expression of PDPN was also higher in stromal cells in comparison with cancer cells. In addition, positive correlation was observed between expression of Twist and PDPN in stromal cells of IDC (r=0.267; p<0.05).
The relationship between the higher expression of Twist in both cancer and stromal cells and shorter patients' survival indicates Twist as a potential useful prognostic marker in IDC. Positive correlation of Twist and PDPN expression may indicate the role of PDPN in EMT in IDC.
背景/目的:由于参与上皮-间质转化(EMT)的转录因子(如Twist)被激活,可观察到上皮标志物受到抑制,间充质标志物表达增加。癌相关成纤维细胞的特异性标志物之一是血小板源性生长因子(PDPN)——一种粘蛋白型膜糖蛋白。本研究旨在探讨Twist和PDPN在浸润性导管癌(IDC)病例中的mRNA和蛋白质水平的定位及表达强度,及其与患者临床病理数据的相关性。
本研究包括80个石蜡包埋的IDC块和11个液氮冷冻的IDC片段形式的存档材料。使用光学显微镜和用于评估核表达的半定量量表或用于评估细胞质表达的免疫反应量表(IRS)评估Twist和PDPN的免疫组化表达。使用激光显微切割从冷冻的IDC片段中分离材料(分别从癌细胞和基质细胞中分离),并用于进行实时PCR。
与癌细胞相比,基质细胞中Twist的表达更高。对患者生存率的分析表明,癌细胞中Twist的高表达与总生存时间缩短和无事件生存时间缩短相关。与癌细胞相比,基质细胞中PDPN的表达也更高。此外,在IDC的基质细胞中观察到Twist和PDPN的表达之间存在正相关(r=0.267;p<0.05)。
癌细胞和基质细胞中Twist的高表达与患者较短的生存期之间的关系表明Twist是IDC中一种潜在有用的预后标志物。Twist和PDPN表达的正相关可能表明PDPN在IDC的EMT中发挥作用。