title: Invasion of uterine cervical squamous cell carcinoma cells is facilitated by locoregional interaction with cancer-associated fibroblasts via activating transforming growth factor-beta.
abstract: OBJECTIVE
  Local invasion is a common pattern of spread in uterine cervical squamous cell carcinoma (CSCC). Although transforming growth factor-beta (TGF-β) facilitates invasion of various types of cancer cells, the role of the TGF-β pathway in CSCC is unclear. In this study, we analyzed the role of TGF-β signaling in the progression of CSCC.
  
  
  METHODS
  Immunohistochemistry was used to examine the expression of TGF-β pathway molecules in 67 CSCC samples with clinicopathological data. Activation of the TGF-β pathway was investigated following co-culture of CSCC cells and cervical cancer-associated fibroblasts (CCAFs).
  
  
  RESULTS
  Clinicopathological analysis of CSCC samples revealed that prominent expression of TGF-β receptor-2 was more frequent in CSCC with lymphovascular space invasion (LVSI) than without LVSI (p < 0.01). Lymph node metastasis was more frequent in cases in which phosphorylated SMAD3 (pSMAD3) was localized exclusively at the boundary of tumor clusters (n = 9, p < 0.05). Recombinant TGF-β1 increased pSMAD3 expression and enhanced cellular invasion (p < 0.005) in CSCC cells, which was attenuated by an inhibitor of the TGF-β receptor (p < 0.005). Enhanced pSMAD3 expression and invasion was also observed when conditioned media from CSCC cells co-cultured with CCAFs were administered. Luciferase assays showed that this medium contained a large amount of active TGF-β. Along with TGF-β activation, thrombospondin-1 was upregulated in both CSCC cells and CCAFs, while thrombospondin-1 silencing in either CSCC cells or CCAFs repressed the activity of TGF-β. Thrombospondin-1 was prominently expressed in cases with pSMAD3 boundary staining (p < 0.05).
  
  
  CONCLUSIONS
  These results suggest that interaction between CSCC cells and surrounding CCAFs activates TGF-β via thrombospondin-1 secretion to facilitate CSCC invasion.
authors.affiliation: Department of Gynecology and Obstetrics, Kyoto University Graduate School of Medicine, Japan.
authors.forename: Michikazu
authors.initials: M
authors.lastname: Nagura
authors.affiliation: Department of Gynecology and Obstetrics, Kyoto University Graduate School of Medicine, Japan.
authors.forename: Noriomi
authors.initials: N
authors.lastname: Matsumura
authors.affiliation: Department of Gynecology and Obstetrics, Kyoto University Graduate School of Medicine, Japan. Electronic address: babatsu@kuhp.kyoto-u.ac.jp.
authors.forename: Tsukasa
authors.initials: T
authors.lastname: Baba
authors.affiliation: Department of Gynecology and Obstetrics, Kyoto University Graduate School of Medicine, Japan.
authors.forename: Ryusuke
authors.initials: R
authors.lastname: Murakami
authors.affiliation: Department of Gynecology and Obstetrics, Kyoto University Graduate School of Medicine, Japan.
authors.forename: Budiman
authors.initials: B
authors.lastname: Kharma
authors.affiliation: Department of Gynecology and Obstetrics, Kyoto University Graduate School of Medicine, Japan.
authors.forename: Junzo
authors.initials: J
authors.lastname: Hamanishi
authors.affiliation: Department of Gynecology and Obstetrics, Kyoto University Graduate School of Medicine, Japan.
authors.forename: Ken
authors.initials: K
authors.lastname: Yamaguchi
authors.affiliation: Department of Gynecology and Obstetrics, Kyoto University Graduate School of Medicine, Japan.
authors.forename: Kaoru
authors.initials: K
authors.lastname: Abiko
authors.affiliation: Department of Gynecology and Obstetrics, Kyoto University Graduate School of Medicine, Japan.
authors.forename: Masafumi
authors.initials: M
authors.lastname: Koshiyama
authors.affiliation: Department of Obstetrics and Gynecology, Kinki University School of Medicine, Japan.
authors.forename: Masaki
authors.initials: M
authors.lastname: Mandai
authors.affiliation: Department of Obstetrics and Gynecology, Kawasaki Medical University, Japan.
authors.forename: Takuya
authors.initials: T
authors.lastname: Murata
authors.affiliation: Division of Gynecologic Oncology, Department of Obstetrics and Gynecology, Duke University Medical Center, Durham, NC, USA.
authors.forename: Susan K
authors.initials: SK
authors.lastname: Murphy
authors.affiliation: Department of Gynecology and Obstetrics, Kyoto University Graduate School of Medicine, Japan.
authors.forename: Ikuo
authors.initials: I
authors.lastname: Konishi
mesh_terms: D002294:Carcinoma, Squamous Cell / Q000378:metabolism / Q000473:pathology*; D002450:Cell Communication / Q000502:physiology*; D005260:Female; D005347:Fibroblasts / Q000378:metabolism / Q000473:pathology*; D006801:Humans; D008875:Middle Aged; D009361:Neoplasm Invasiveness; D009367:Neoplasm Staging; D015398:Signal Transduction; D016212:Transforming Growth Factor beta / Q000378:metabolism*; D002583:Uterine Cervical Neoplasms / Q000378:metabolism / Q000473:pathology*
chemical_list: D016212:Transforming Growth Factor beta
keywords: Cancer associated fibroblasts; Cervical cancer; Invasion; TGF-beta