Cancer associated ﬁbroblasts are distinguishable from peri‑tumor ﬁbroblasts by biological characteristics in TSCC

Ba,Pengfei; Zhang,Xiaojuan; Yu,Miao; Li,Linxia; Duan,Xiaoyu; Wang,Mingying; Lv,Shuyan; Fu,Guo; Yang,Pishan; Yang,Chengzhe; Sun,Qinfeng

doi:10.3892/ol.2019.10556

Cancer associated ﬁbroblasts are distinguishable from peri‑tumor ﬁbroblasts by biological characteristics in TSCC

Authors:
- Pengfei Ba
- Xiaojuan Zhang
- Miao Yu
- Linxia Li
- Xiaoyu Duan
- Mingying Wang
- Shuyan Lv
- Guo Fu
- Pishan Yang
- Chengzhe Yang
- Qinfeng Sun
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Affiliations: Department of Periodontology, School of Stomatology, Shandong University, Jinan, Shandong 250012, P.R. China, Department of Stomatology, Beijing Luhe Hospital Affiliated to Capital Medical University, Beijing 101149, P.R. China, Department of Stomatology, Weifang People's Hospital, Weifang, Shandong 261000, P.R. China, Department of Stomatology, Affiliated Hospital of Jining Medical University, Jining, Shandong 272000, P.R. China, National Engineering Laboratory, WeGo Group Co., Ltd., Weihai, Shandong 264200, P.R. China, Department of Periodontology, Weihai Stomatological Hospital, Weihai, Shandong 264200, P.R. China, Department of Oral and Maxillofacial Surgery, Qilu Hospital, Institute of Stomatology, Shandong University, Jinan, Shandong 250012, P.R. China
Published online on: July 4, 2019 https://doi.org/10.3892/ol.2019.10556
Pages: 2484-2490
Copyright: © Ba et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The aim of the present study was to investigate the differential biological characteristics between cancer‑associated ﬁbroblasts (CAFs) and peri‑tumor ﬁbroblasts (PTFs) in tongue squamous cell carcinoma (TSCC). The primary CAFs and PTFs from TSCC were obtained and purified. Cell morphology was observed, and the expression of α‑smooth muscle actin (α‑SMA), vimentin and cytokeratin 19 (CK19) was detected by immunohistochemistry (IHC). The percentage of α‑SMA positive cells in CAFs and PTFs was calculated separately, and α‑SMA expression was further confirmed by western blot analysis. Cell viability and the expression of matrix metalloproteinase 2 (MMP2), stromal cell derived factor1 (SDF‑1) and transforming growth factor β1 (TGFβ1) in the purified fibroblasts was detected separately. CAFs and PTFs were attained and purified. Compared with PTFs, CAFs were long‑fusiform shaped cells with reduced cytoplasm and variable size. CAFs crowded together in a disorderly manner when the cell density was increased, but this phenomenon did not occur with PTFs. IHC results verified that there was no significant difference between CAFs and PTFs in the percentage of cells staining positive for CK19 and vimentin (P>0.05); the percentage of positive staining cells for α‑SMA in CAFs was significantly higher compared with that in PTFs (P<0.001) Western blot analysis showed that α‑SMA expression in CAFs was 4.3‑fold higher compared with that in PTFs (P<0.001). A Cell Counting Kit‑8 assay indicated that the viability of CAFs increased significantly compared with that in the PTFs (P<0.05). Reverse transcription‑quantitative polymerase chain reaction and ELISA analysis showed that the expression of MMP2, SDF‑1 and TGF β1 in CAFs was higher compared with that in PTFs (P<0.05). CAFs are distinguishable from PTFs with respect to their morphology, cellular phenotype, cell viability and pro‑carcinogenic cytokine expression.

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