Elsevier

Gynecologic Oncology

Volume 136, Issue 1, January 2015, Pages 104-111
Gynecologic Oncology

Invasion of uterine cervical squamous cell carcinoma cells is facilitated by locoregional interaction with cancer-associated fibroblasts via activating transforming growth factor-beta

https://doi.org/10.1016/j.ygyno.2014.11.075Get rights and content

Highlights

  • TGF-β pathway is distinguished as a driver initiating invasion of cervical squamous cell carcinoma.

  • TGF-β is activated through interaction of squamous cell carcinoma cells and cancer associated fibroblasts at tumor invasive front.

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.

Introduction

Despite widespread vaccination against human papilloma virus and periodic cancer screening, cervical cancer remains one of the highest ranking diseases causing mortality in women, and new strategies to treat this disease are urgently needed. Squamous cell carcinoma, the most common histological subtype of cervical cancer, spreads principally by migrating into the lymphatics or by invading adjacent soft tissues. Eradication of locoregional lesions is critical but is not attainable in cases with tumor extension into the urinary tract and/or rectum for fear of impairing urinary or bowel function. Even if the extension is more limited, it is also difficult to obtain cancer-free-margins, leading to pelvic recurrence from residual cancer cells. Obstruction of the ureter(s) is not trivial and is observed in 55.8% of advanced cases [1]. It is, therefore, important to develop effective treatments for invasive extension of this disease, and to this end, it is essential to further elucidate the mechanisms of cervical cancer invasion.

Transforming growth factor-beta (TGF-β) is currently known to promote cancer invasion [2]. TGF-β serves as a potent growth inhibitor for normal epithelial cells [3], but malignant transformed cells acquire resistance to the growth inhibitory effect of TGF-β [4]. Moreover, as breast cancer advances, TGF-β acts as a promoter by inducing epithelial to mesenchymal transition (EMT), invasion, and metastasis [5], [6]. In gynecologic cancers, activated TGF-β signaling promotes peritoneal dissemination of ovarian cancer [7]. Thus, TGF-β is widely recognized as a key molecule that drives cancer cell progression, but little is known about the role of TGF-β in uterine cervical cancer.

Growing evidence has shown that the stroma around cancer cells plays an important role in cancer progression. Cancer-associated fibroblasts (CAFs) exhibit morphological phenotypes of myofibroblasts and are known to promote cancer progression through interactions with adjacent cancer cells. TGF-β is a key cytokine mediating such interactions [8], [9]. However, we presently lack clarification regarding the role of TGF-β in these interactions in cervical squamous cell carcinoma (CSCC).

In this study, we analyze the role of TGF-β signaling in the progression of CSCC. We show that activation of TGF-β induced by the interaction between CSCC cells and CAFs plays a key role in the initiation of tumor metastasis.

Section snippets

Tissue samples and immunohistochemistry

Tissue samples and clinicopathological information were collected from sixty-seven patients (median age, 54 +/− 23 years) with stage IB–IIB CSCC who underwent radical hysterectomy or trachelectomy in Kyoto University Hospital from January 2003 to July 2010 with written consent under the approval of the ethics committee. Patient characteristics are described in Table 1.

Immunohistochemical staining was performed for six molecules: TGF-β, TGF-β receptor type 1 (TGFBR1), TGF-β receptor type 2

Clinicopathological analysis of CSCC in association with the TGF-β pathway

A total of 67 patients were treated for locally advanced CSCC (IB1; n = 38, IB2; n = 6, IIA1; n = 5, IIA2; n = 2, and IIB; n = 16). The 67 cases included 22 with lymph node metastasis and two with distant metastasis. Patients with lymphovascular space invasion (LVSI) or lymph node metastasis (LNM) exhibited worse clinical outcomes than those without these characteristics (p < 0.05 for each, Table 1). Immunohistochemical staining was conducted for TGF-β, TGFBR1, TGFBR2, pSMAD3, α-SMA, and TSP1. TGF-β was

Discussion

TGF-β is known to play various roles in malignant progression. It inhibits growth of normal epithelial cells, and conversely, it promotes EMT, invasion and metastasis in cancer cells, thus eluding tumor-suppressive effects [2], [3], [4]. With regard to CSCC, the role of TGF-β signaling is more complicated and not well understood. TGF-β pathway functions in an inhibitory manner for pelvic lymph node metastasis [13], while TGF-β1 expression increases through progression from cervical

Conflict of interest statement

All authors declare that they have no conflicts of interest to disclose.

References (21)

There are more references available in the full text version of this article.

Cited by (20)

  • Tumor-Secreted Exosomal lncRNA POU3F3 Promotes Cisplatin Resistance in ESCC by Inducing Fibroblast Differentiation into CAFs

    2020, Molecular Therapy Oncolytics
    Citation Excerpt :

    In contrast to the above reports implicating the direct enhancing effects of lncRNA POU3F3 on cell survival, our data identified that lncRNA POU3F3 can indirectly affect tumor growth by acting on the TME. CAFs occupy a central position in the TME and have been reported to create suitable conditions for the progression of cancer cells.38,39 Furthermore, they are often the key factors that foster the resistance to therapy, whether it is chemotherapy, RT, or targeted approaches.40

  • MiR-449b-5p regulates cell proliferation, migration and radioresistance in cervical cancer by interacting with the transcription suppressor FOXP1

    2019, European Journal of Pharmacology
    Citation Excerpt :

    Cervical cancer (CC) is acknowledged as one of the most common gynecologic tumors (Jain et al., 2008). Although the vaccination against human papillomavirus and surgical treatment have been developed and improved (Haggar et al., 2012; Lea and Lin, 2012), the mortality of CC patients is still quite high (Nagura et al., 2015; Torre et al., 2015). Therefore, finding novel effective therapeutic method for cervical cancer is urgent in need.

  • Cutaneous Metastasis via Surgical Planes After Lumbar Spinal Reconstruction for Spinal Metastatic Disease

    2019, World Neurosurgery
    Citation Excerpt :

    Squamous cell carcinoma is the most frequent subtype, accounting for 70%−80% of cervical cancers.3 This invasive cancer frequently metastasizes and can spread hematogenously, through lymphatics, or via direct extension.4 Though uncommon, CSCC has been reported to metastasize locally to drain sites and postpartum episiotomy scars.5-7

  • Negative roles of B7-H3 and B7-H4 in the microenvironment of cervical cancer

    2018, Experimental Cell Research
    Citation Excerpt :

    We showed that expression of IL-10 was higher in HSIL and cervical cancer than in normal cervix, consistent with the findings of previous studies [30,31]; however, no significant differences in TGF-β1 expression were found in these three groups, possibly related to its versatility in various phases of tumor progression. The analysis of IL-10/TGF-β1 expression and pathological indices suggested that IL-10 was related to tumor differentiation, and TGF-β1 was related to LNM, in accordance with its effects on invasiveness and migration [12,13]. We performed Pearson r tests and found that expression of B7-H3/B7-H4 were positively correlated with IL-10/TGF-β1 except for B7-H4 and TGF-β1, and the likely cause was that B7-H4 was distributed in cervical tumor cells and TGF-β1 was in tumor stroma, but the area of TMA was limited.

  • The Interaction Between Human Papillomaviruses and the Stromal Microenvironment

    2016, Progress in Molecular Biology and Translational Science
    Citation Excerpt :

    Conditioned media from cervical cancer-derived CAFs increase HPV16-immortalized keratinocyte invasion.194 Coculture of cervical cancer cells with cervical CAFs increases TGFβ1 and TSP-1 secretion.197 CAFs isolated from cervical cancers secrete more IL6, IL8, and VEGF and have more STAT3 activation than normal fibroblasts.198

View all citing articles on Scopus
View full text