Zhou Bin, Chen Wei-Liang, Wang You-Yuan, Lin Zhao-Yu, Zhang Da-Ming, Fan Song, Li Jin-Song
Department of Oral and Maxillofacial Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China.
J Oral Pathol Med. 2014 Sep;43(8):585-92. doi: 10.1111/jop.12172. Epub 2014 Mar 20.
Lymph node metastasis is a prominent clinical feature of tongue squamous cell carcinoma (TSCC) and is associated with a higher mortality rate. Carcinoma-associated fibroblasts (CAFs), a major component of the tumor microenvironment (TME), play an important role in tumor progression, and are associated with a poor prognosis. The aim of this study was to examine the role of CAFs in promoting the invasion of TSCC through the epithelial-to-mesenchymal transition (EMT).
A series of matched CAF and normal fibroblast (NF) pairs were assessed for cell morphology and for the expression of alpha smooth muscle actin (α-SMA), stromal cell-derived factor-1 (SDF1), fibroblast-activating protein (FAP), vimentin, and cytokeratin (CK) markers. Transwell assays, Western blot analysis, reverse transcription-PCR, and immunofluorescence staining were used to assess the role of CAFs, as compared to that of NFs, in promoting proliferation, migration, invasion, and EMT in TSCC.
Both CAF and NF primary cultures expressed vimentin but not CK. CAFs showed significantly higher α-SMA protein levels, SDF1 secretion, and mRNA levels of α-SMA, SDF1, and FAP. We also found that co-culture with CAFs enhanced the proliferation and invasion of SCC9 cells. Moreover, co-culture with CAFs induced upregulation of the EMT markers fibronectin and vimentin, downregulation of E-cadherin, and enhanced invasion in SCC9 cells.
These results suggest that CAFs induce EMT marker expression and functional changes in TSCCs.
淋巴结转移是舌鳞状细胞癌（TSCC）的一个显著临床特征，且与较高的死亡率相关。癌相关成纤维细胞（CAFs）是肿瘤微环境（TME）的主要组成部分，在肿瘤进展中起重要作用，并与预后不良相关。本研究的目的是探讨CAFs通过上皮-间质转化（EMT）促进TSCC侵袭的作用。
评估一系列配对的CAF和正常成纤维细胞（NF）对的细胞形态以及α平滑肌肌动蛋白（α-SMA）、基质细胞衍生因子-1（SDF1）、成纤维细胞激活蛋白（FAP）、波形蛋白和细胞角蛋白（CK）标志物的表达。与NFs相比，采用Transwell试验、蛋白质印迹分析、逆转录-PCR和免疫荧光染色来评估CAFs在促进TSCC增殖、迁移、侵袭和EMT中的作用。
CAF和NF原代培养物均表达波形蛋白，但不表达CK。CAFs显示出显著更高的α-SMA蛋白水平、SDF1分泌以及α-SMA、SDF1和FAP的mRNA水平。我们还发现与CAFs共培养增强了SCC9细胞的增殖和侵袭。此外，与CAFs共培养诱导了EMT标志物纤连蛋白和波形蛋白的上调、E-钙黏蛋白的下调，并增强了SCC9细胞的侵袭。
这些结果表明CAFs诱导TSCC中EMT标志物的表达和功能变化。
