Involvement of cysteine-rich protein 61 in the epidermal growth factor-induced migration of human anaplastic thyroid cancer cells

Hölting T, Siperstein AE, Clark OH, Duh Q-Y. Epidermal growth factor (EGF)- and transforming growth factor alpha-stimulated invasion and growth of follicular thyroid cancer cells can be blocked by antagonism to the EGF receptor and tyrosine kinase in vitro. Eur J Endocrinol 1995;132:229–35. ISSN 0804–4643 We have shown recently that epidermal growth factor (EGF) enhanced invasion and growth of differentiated thyroid cancer cells in vitro and in vivo. The present study analyzed the effects of transforming growth factor alpha (TGF-α) on invasion and growth of a follicular thyroid cancer cell line (FTC133) and whether blocking the EGF receptor by a monoclonal antibody (Mab528) or blocking the tyrosine kinase of the receptor by genistein abolished the EGF- and TFG-α-mediated effects. Growth and invasion (penetration of 8-μm pore polycarbonate membranes coated with Matrigel) were determined by the dimethylthiazol-diphenyltetrazolium bromide assay. Epidermal growth factor (10 ng/l) stimulated invasion of FTC133 by 42% and TGF-α (10 ng/l) stimulated invasion of FTC133 by 27% (p < 0.02). Both growth factors also enhanced growth by 62% (EGF) and 30% (TGF-α) (p < 0.003). Epidermal growth factor receptor antibodies (1 μg/ml) abolished EGF-mediated stimulation of invasion and growth completely and that of TGF-α by 93%. At 100 ng/ml genistein reversed EGF and TGF-α stimulation, and at 1 μg/ml it inhibited invasion (27%) and growth (40%) of unstimulated FTC133 (p < 0.02). We conclude that TGF-α stimulates invasion and growth of follicular thyroid cancer by binding to the EGF receptors, that EGF- and TGF-α-mediated effects can be blocked by antagonism to the EGF receptor and to tyrosine kinase, and that genistein not only neutralized EGF and TGF-α effects but also inhibited invasion and growth of unstimulated FTC133. Therefore, tyrosine kinase activity via other signalling systems must be crucial for basal invasion and growth of follicular thyroid cancer cells in culture. Thomas Hölting, Department of Surgery, University of Heidelberg, Im Neuenheimer Feld 110, 69120 Heidelberg, Germany

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CLM3, a Multitarget Tyrosine Kinase Inhibitor With Antiangiogenic Properties, Is Active Against Primary Anaplastic Thyroid Cancer In Vitro and In Vivo

The Journal of Clinical Endocrinology & Metabolism ◽

10.1210/jc.2013-2321 ◽

2014 ◽

Vol 99 (4) ◽

pp. E572-E581 ◽

Cited By ~ 24

Author(s):

Alessandro Antonelli ◽

Guido Bocci ◽

Poupak Fallahi ◽

Concettina La Motta ◽

Silvia Martina Ferrari ◽

...

Keyword(s):

Epidermal Growth Factor Receptor ◽

Thyroid Cancer ◽

Growth Factor ◽

Epidermal Growth Factor ◽

Tyrosine Kinase ◽

Cell Line ◽

Growth Factor Receptor ◽

Anaplastic Thyroid Cancer ◽

Antiangiogenic Activity ◽

Epidermal Growth

Context and Objective: We have studied the antitumor activity of a pyrazolo[3,4-d]pyrimidine compound (CLM3) proposed for a multiple signal transduction inhibition [including the RET tyrosine kinase, epidermal growth factor receptor, and vascular endothelial growth factor (VEGF) receptor and with antiangiogenic activity] in primary anaplastic thyroid cancer (ATC) cells, in the human cell line 8305C (undifferentiated thyroid cancer), and in an ATC-cell line (AF). Design and Main Outcome Measures: CLM3 was tested in primary ATC cells at the concentrations of 5, 10, 30, and 50 μM; in 8305C cells, in AF cells, at 1, 5, 10, 30, 50, or 100 μM; and in AF cells in CD nu/nu mice. Results: CLM3 significantly inhibited the proliferation of 8305C and AF cells, also inducing apoptosis. A significant reduction of proliferation with CLM3 in ATC cells (P < .01, ANOVA) was shown. CLM3 increased the percentage of apoptotic ATC cells dose dependently (P < .001, ANOVA) and inhibited migration (P < .01) and invasion (P < .001). The AF cell line was injected sc in CD nu/nu mice, and tumor masses became detectable 15 days later. CLM3 (50 mg/kg per die) significantly inhibited tumor growth (starting 16 d after the beginning of treatment). CLM3 significantly decreased the VEGF-A expression and microvessel density in AF tumor tissues. Furthermore, CLM3 inhibited epidermal growth factor receptor, AKT, and ERK1/2 phosphorylation and down-regulated cyclin D1 in 8305C and AF cells. Conclusions: The antitumor and antiangiogenic activity of a pyrazolo[3,4-d]pyrimidine compound (CLM3) is very promising in anaplastic thyroid cancer, opening the way to a future clinical evaluation.

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