title: Cancer-associated fibroblasts in a human HEp-2 established laryngeal xenografted tumor are not derived from cancer cells through epithelial-mesenchymal transition, phenotypically activated but karyotypically normal.
abstract: Cancer-associated fibroblasts (CAFs) play a crucial role in cancer progression and even initiation. However, the origins of CAFs in various cancer types remain controversial, and one of the important hypothesized origins is through epithelial-mesenchymal transition (EMT) from cancer cells. In this study, we investigated whether the HEp-2 laryngeal cancer cells are able to generate CAFs via EMT during tumor formation, which is now still unknown. The laryngeal xenografted tumor model was established by inoculating the HEp-2 laryngeal cancer cell line in nude mice. Primary cultured CAFs from the tumor nodules and matched normal fibroblasts (NFs) from the adjacent connective tissues were subcultured, purified, and verified by immunofluorescence. Migration, invasion, and proliferation potentials were compared between the CAFs and NFs. A co-culture of CAFs with HEp-2 cells and a co-injection of CAFs with HEp-2 cells in nude mice were performed to examine the cancer-promoting potential of CAFs to further verify their identity. Karyotypic analyses of the CAFs, NFs, and HEp-2 cells were conducted. A co-culture of NFs with HEp-2 cells was also performed to examine the expression of activated markers of CAFs. A pathological examination confirmed that the laryngeal xenografted tumor model was successfully established, containing abundant CAFs. Immunocytochemical staining verified the purities and identities of the CAFs and NFs. Although the CAFs manifested higher migration, invasion, proliferation, and cancer-promoting capacities compared with the NFs, an analysis of chromosomes revealed that both the CAFs and NFs showed typical normal mouse karyotypes. In addition, the NFs co-cultured with HEp-2 cells did not show induced expressions of activated markers of CAFs. Our findings reveal that the CAFs in the HEp-2 established laryngeal xenografted tumor are not of laryngeal cancer origin but of mouse origin, indicating that the HEp-2 laryngeal cancer cells cannot generate their own CAFs via EMT in this model.
authors: Department of Otolaryngology, Shanghai Municipal Hospital of Traditional Chinese Medicine, Shanghai, People's Republic of China; Department of Otolaryngology, the Second Affiliated Hospital of Jiaxing University, Jiaxing, Zhejiang Province, People's Republic of China.
authors: Mei
authors: M
authors: Wang
authors: Department of Otolaryngology-Head and Neck Surgery, Eye, Ear, Nose and Throat Hospital of Fudan University, Shanghai, People's Republic of China.
authors: Chun-Ping
authors: CP
authors: Wu
authors: Department of Otolaryngology, Shanghai Municipal Hospital of Traditional Chinese Medicine, Shanghai, People's Republic of China.
authors: Jun-Yan
authors: JY
authors: Pan
authors: Department of Otolaryngology, Shanghai Municipal Hospital of Traditional Chinese Medicine, Shanghai, People's Republic of China.
authors: Wen-Wei
authors: WW
authors: Zheng
authors: Department of Otolaryngology, Shanghai Municipal Hospital of Traditional Chinese Medicine, Shanghai, People's Republic of China.
authors: Xiao-Juan
authors: XJ
authors: Cao
authors: Department of Otolaryngology, the Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, Zhejiang Province, People's Republic of China.
authors: Guo-Kang
authors: GK
authors: Fan
mesh_terms: D000818:Animals; D002460:Cell Line; D002465:Cell Movement; D049109:Cell Proliferation; D018920:Coculture Techniques; D058750:Epithelial-Mesenchymal Transition; D005347:Fibroblasts / Q000378:metabolism*; D064593:Heterografts; D006801:Humans; D059785:Karyotype; D007822:Laryngeal Neoplasms / Q000378:metabolism / Q000473:pathology*; D008819:Mice, Nude; D009361:Neoplasm Invasiveness; D010641:Phenotype