Cancer-associated fibroblasts induce epithelial-to-mesenchymal transition in tongue squamous cell carcinoma

Abstract

Abstract: PURPOSE: To examine the role of cancer-associated fibroblasts (CAFs) in promoting invasion of tongue squamous cell carcinoma (TSCC) and the possible mechanism. METHODS: A series of matched pairs of CAFs and normal fibroblasts (NFs) were primarily cultured in collagenase digestion. Identified markers of alpha smooth muscle actin (α-SMA), stromal cell-derived factor-1(SDF1), vimentin and cytokeratin (CK) were observed. Transwell assay, Western blot, reverse transcription-PCR and immunofluorescence staining were carried out to assess the role of CAFs in promoting migration, invasion and inducing epithelial to mesenchymal transition (EMT) of SCC9, compared with NFs. RESULTS: Both primarily cultured CAFs and NFs expressed vimentin, but not CK. CAFs showed significantly higher α-SMA expression and SDF1 secretion, as well as mRNA level of α-SMA and SDF1. Co-culture with CAFs induced up-regulation of EMT markers (fibronectin and vimentin) and down-regulation of E-cadherin of SCC9, as well as enhanced invasion. CONCLUSION: Collagenase digestion is a reliable way to primary culture of CAFs. CAFs may induce EMT markers and functional changes in SCC9. Supported by National Natural Science Foundation of China (81172563).

Key words: Cancer-associated fibroblast, Tumor microenvironment, Tongue squamous cell carcinoma, Epithelial-mesenchymal transition

CLC Number:

R739.8

ZHOU Bin, CHEN Wei-liang, WANG You-yuan, LIN Zhao-yu, ZHANG Da-ming, FAN Song. Cancer-associated fibroblasts induce epithelial-to-mesenchymal transition in tongue squamous cell carcinoma[J]. China Journal of Oral and Maxillofacial Surgery, 2014, 12(3): 198-204.

References

【1】Sun L, Yao Y, Liu B, et al. MiR-200b and miR-15b regulate chemotherapy-induced epithelial-mesenchymal transition in human tongue cancer cells by targeting BMI1 . Oncogene, 2012, 31(4): 432-445.
【2】Joyce JA, Pollard JW. Microenvironmental regulation of metastasis . Nat Rev Cancer, 2009, 9(4): 239-252.
【3】Zhang J, Liu J. Tumor stroma as targets for cancer therapy . Pharmacol Ther, 2013, 137(2): 200-215.
【4】Potenta S, Zeisberg E, Kalluri R. The role of endothelial-to-mesenchymal transition in cancer progression . Br J Cancer, 2008, 99(9): 1375-1379.
【5】Liu Y, Hu T, Shen J, et al. Separation, cultivation and biological characteristics of oral carcinoma-associated fibroblasts . Oral Dis, 2006, 12(4): 375-380.
【6】Hanna E, Quick J, Libutti SK. The tumour microenvironment: a novel target for cancer therapy . Oral Dis, 2009, 15(1): 8-17.
【7】Fujii N, Shomori K, Shiomi T, et al. Cancer-associated fibroblasts and CD163-positive macrophages in oral squamous cell carcinoma: their clinicopathological and prognostic significance . J Oral Pathol Med, 2012, 41(6): 444-451.
【8】Bello IO, Vered M, Dayan D, et al. Cancer-associated fibroblasts, a parameter of the tumor microenvironment, overcomes carcinoma-associated parameters in the prognosis of patients with mobile tongue cancer . Oral Oncol, 2011, 47(1): 33-38.
【9】Vered M, Dobriyan A, Dayan D, et al. Tumor-host histopathologic variables, stromal myofibroblasts and risk score,are significantly associated with recurrent disease in tongue cancer . Cancer Sci, 2010, 101(1): 274-280.
【10】Schoppmann SF, Berghoff A, Dinhof C, et al. Podoplanin-expressing cancer-associated fibroblasts are associated with poor prognosis in invasive breast cancer . Breast Cancer Res Treat, 2012, 134(1): 237-244.【11】Herrera M, Herrera A, Dominguez G, et al. Cancer-associated fibroblast and M2 macrophage markers together predict outcome in colorectal cancer patients . Cancer Sci, 2013, 104(4): 437-444.
【12】Orimo A, Gupta PB, Sgroi DC, et al. Stromal fibroblasts present in invasive human breast carcinomas promote tumor growth and angiogenesis through elevated SDF-1/CXCL12 secretion . Cell, 2005, 121(3): 335-348.
【13】Lin J, Liu C, Ge L, et al. Carcinoma-associated fibroblasts promotes the proliferation of a lingual carcinoma cell line by secreting keratinocyte growth factor . Tumour Biol, 2011, 32(3): 597-602.
【14】Sobral LM, Bufalino A, Lopes MA, et al. Myofibroblasts in the stroma of oral cancer promote tumorigenesis via secretion of activin A . Oral Oncol, 2011, 47(9): 840-846.
【15】Wu MH, Hong HC, Hong TM, et al. Targeting galectin-1 in carcinoma-associated fibroblasts inhibits oral squamous cell carcinoma metastasis by downregulating MCP-1/CCL2 expression . Clin Cancer Res, 2011, 17(6): 1306-1316.
【16】Hinsley EE, Hunt S, Hunter KD, et al. Endothelin-1 stimulates motility of head and neck squamous carcinoma cells by promoting stromal-epithelial interactions. Int J Cancer, 2012, 130(1): 40-47.
【17】Xu J, Lu Y, Qiu S, et al. A novel role of EMMPRIN/CD147 in transformation of quiescent fibroblasts to cancer-associated fibroblasts by breast cancer cells . Cancer Lett, 2013, 335(2): 380-386.
【18】Kellermann MG, Sobral LM, da Silva SD, et al. Mutual paracrine effects of oral squamous cell carcinoma cells and normal oral fibroblasts: induction of fibroblast to myofibroblast transdifferentiation and modulation of tumor cell proliferation . Oral Oncol, 2008, 44(5): 509-517.
【19】Thiery JP, Acloque H, Huang RY, et al. Epithelial-mesenchymal transitions in development and disease . Cell, 2009, 139(5): 871-890.
【20】Vered M, Dayan D, Yahalom R, et al. Cancer-associated fibroblasts and epithelial-mesenchymal transition in metastatic oral tongue squamous cell carcinoma. Int J Cancer,2010,127(6): 1356-1362.
【21】Lebret SC, Newgreen DF, Thompson EW, et al. Induction of epithelial to mesenchymal transition in PMC42-LA human breast carcinoma cells by carcinoma-associated fibroblast secreted factors . Breast Cancer Res, 2007, 9(1): R19.
【22】Soon P, Kim E, Pon CK, et al. Breast cancer-associated fibroblasts induce epithelial-to-mesenchymal transition in breast cancer cells . Endocr Relat Cancer, 2013, 20(1): 1-12.
【23】Giannoni E, Bianchini F, Masieri L, et al. Reciprocal activation of prostate cancer cells and cancer-associated fibroblasts stimulates epithelial-mesenchymal transition and cancer stemness. Cancer Res, 2010, 70(17): 6945-6956.
【24】DudÁs J, Bitsche M, Schartinger V, et al. Fibroblasts produce brain-derived neurotrophic factor and induce mesenchymal transition of oral tumor cells . Oral Oncol, 2011, 47(2): 98-103.
【25】Daly AJ, McIlreavey L, Irwin CR. Regulation of HGF and SDF-1 expression by oral fibroblasts-implications for invasion of oral cancer . Oral Oncol, 2008, 44(7): 646-651.
【26】Onoue T, Uchida D, Begum NM, et al. Epithelial-mesenchymal transition induced by the stromal cell-derived factor-1/CXCR4 system in oral squamous cell carcinoma cells . Int J Oncol, 2006, 29(5): 1133-1138.

[1]	HAN Lin-zi, ZHOU Jian-hua, ZHAO Qian, DONG Lei, CHEN Zheng-gang, QIU Jian-zhong, YUAN Rong-tao. The regulation and mechanism of cancer-associated fibroblasts on tumor-infiltrating immune cells in head and neck squamous cell carcinoma [J]. China Journal of Oral and Maxillofacial Surgery, 2023, 21(2): 119-124.
[2]	SHUI Wen-jun, GAO Jin-xing, REN Lei. Effect of MMP-9 inhibitor on invasion and migration ability of human tongue squamous cell carcinoma SCC-25 cell and its mechanism [J]. China Journal of Oral and Maxillofacial Surgery, 2021, 19(5): 424-428.
[3]	YE Chen, CAI Yun, ZHAO Xiao-mei, ZHANG Chen-ping, ZHAO Nai-qing. Prognostic factors of oral tongue squamous cell carcinoma: a retrospective study of 638 cases [J]. China Journal of Oral and Maxillofacial Surgery, 2021, 19(5): 449-455.
[4]	CHEN Zheng-gang, WANG Qi-min, TONG Lei, WANG Yun-ying, XU Xiao-na, WANG Ying, HAN Hong-yu, SHENG Shan-gui, WANG Shao-ru. Effects of silencing Icmt on the proliferation, apoptosis, and cell cycle of CAL-27 and SCC-4 cells [J]. China Journal of Oral and Maxillofacial Surgery, 2021, 19(2): 97-104.
[5]	KONG Xian-chen, JIANG Xiao-dong, LU Xin, YU Wen-yi, XUE Jiao, ZHANG Peng, ZHAO Lu, YUAN Rong-tao. Expression and significance of ribosomal protein L29 in tongue squamous cell carcinoma [J]. China Journal of Oral and Maxillofacial Surgery, 2021, 19(2): 110-115.
[6]	HE Xing-fang, LIN Yan-tong, WANG Shuai, YE Jing-jing, LIU Jie, HUANG Qiu-yu. Changes of swallowing function in 40 patients with tongue squamous cell carcinoma after simultaneous reconstruction: a longitudinal study [J]. China Journal of Oral and Maxillofacial Surgery, 2021, 19(1): 12-17.
[7]	LI Ming-yu, MA Hai-long, YANG Xi, ZHANG Zhi-yuan. The effect of cancer-associated fibroblast in glucose metabolism of tumor cells [J]. China Journal of Oral and Maxillofacial Surgery, 2021, 19(1): 70-75.
[8]	WANG Zi-yu, DING Yu-jie, CHEN Bing, SHEN Hao-chong, MENG Jian, TONG Wei-feng. Inhibitory effect of α-eleostearic acid on tongue squamous cell carcinoma CAL-27 [J]. China Journal of Oral and Maxillofacial Surgery, 2020, 18(6): 490-494.
[9]	ZHOU Guang-ming, LIANG Yan-can, HUANG Zi-xian, ZHANG Bin. Effect of long noncoding RNA FOXD2-AS1 on migration and invasion of tongue squamous cell carcinoma: an in vitro study [J]. China Journal of Oral and Maxillofacial Surgery, 2020, 18(6): 495-500.
[10]	XU Qiao-shi, YANG Xi, MA Hai-long, CHANG Han-yue, ZHANG Chen-ping. Progress on interleukin-8 in epithelial-mesenchymal transition of oral squamous cell carcinoma [J]. China Journal of Oral and Maxillofacial Surgery, 2020, 18(6): 563-566.
[11]	CHEN Yong-ju, CHEN Wei-liang, ZHOU Bin, ZHONG Jiang-long, WANG Yan, CHEN Rui.. Comparison of facial-submental artery island flap for repairing younger and older patients with tongue squamous cell carcinoma [J]. China Journal of Oral and Maxillofacial Surgery, 2020, 18(2): 117-121.
[12]	LIU Guo-xin, WANG Wei-wei, MAI Lian-xi, JIAO Jiu-yang, XIE Shu-le, LIN Zhao-yu. Cisplatin induces epithelial-mesenchymal transition in tongue squamous carcinoma cells and obtains tumor stem cell-like characteristics [J]. China Journal of Oral and Maxillofacial Surgery, 2020, 18(1): 10-16.
[13]	LIN Zhi-li, LIANG Jian-ping, ZHOU Bin. Hypoxia induces epithelial-to-mesenchymal transition of human tongue squamous cell carcinoma via lncRNA-HOTTIP [J]. China Journal of Oral and Maxillofacial Surgery, 2019, 17(1): 26-31.
[14]	LUO Dan, SUN Qiao-zhen, LIU Shao-hua, SUN Jun-wei, WANG Sheng-zhi. Effect of hyperthermia combined with silencing Id-1 gene on proliferation and invasion of tongue squamous cell carcinoma cell line CAL-27 [J]. China Journal of Oral and Maxillofacial Surgery, 2018, 16(6): 488-492.
[15]	MA Li, SHI Bing, ZHANG Dong-sheng, ZHENG Qian. The effect of apoptosis and epithelial-mesenchymal transition during palatal fusion [J]. China Journal of Oral and Maxillofacial Surgery, 2018, 16(6): 493-497.