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MET receptor is a potential therapeutic target in high grade cervical cancer.

Miekus K, Pawlowska M, Sekuła M, Drabik G, Madeja Z, Adamek D, Majka M - Oncotarget (2015)

Bottom Line: MET receptor downregulation also resulted in decreased cyclin D1 and c-myc levels but did not increase apoptosis.Subsequent experiments showed that downregulation of the MET receptor decreased the expression of a key regulator of the epithelial-to-mesenchymal transition, SLUG. and increased the expression of E-cadherin, a hallmark of the epithelial phenotype.Taken together, our results strongly suggest that the MET receptor influences the oncogenic properties of cervical carcinoma cells in vitro and in vivo.

View Article: PubMed Central - PubMed

Affiliation: Department of General Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Cracow, Poland.

ABSTRACT
Cervical cancer is one of the leading causes of death among women suffering from tumors. Current treatment options are insufficient. Here, we investigated the MET receptor as a potential molecular target in advanced cervical cancer. Downregulation of MET receptor expression via RNA interference in different cervical carcinoma cell lines dramatically decreased tumor growth and forced tumor differentiation in vivo. MET receptor silencing also led to a dramatic decrease in cell size and a decrease in proliferation rate under normal and stress conditions. MET receptor downregulation also resulted in decreased cyclin D1 and c-myc levels but did not increase apoptosis. Subsequent experiments showed that downregulation of the MET receptor decreased the expression of a key regulator of the epithelial-to-mesenchymal transition, SLUG. and increased the expression of E-cadherin, a hallmark of the epithelial phenotype. Moreover, MET downregulation impairs expression and signaling of CXCR4 receptor, responsible for invasive phenotype. Taken together, our results strongly suggest that the MET receptor influences the oncogenic properties of cervical carcinoma cells in vitro and in vivo. These findings highlight a unique role of the MET receptor in cervical carcinoma cells and indicate the MET receptor as a potential therapeutic target for advanced cervical carcinoma.

No MeSH data available.


Related in: MedlinePlus

MET receptor downregulation decreases CXCR4 expression and function(A) Immunohistochemical analysis of CXCR4 receptor expression in human tissue. (B) Real-time RT-PCR and flow cytometry analysis revealed significant decreases in MET transcript and protein levels in MET receptor-silenced cells (shMET) relative to controls (wild type, WT, and shLacZ). (C) Western blot analysis of CXCR4 activation. AKT and MAPK phosphorylation was tested after after HGF and SDF-1 stimulation. (D) In chemotaxis assay MET-deficient CC cells showed limited chemotactic activity towards HGF and/or SDF-1 gradient. Western blot and FACS analyses were performed three times with similar results; representative results are shown. Real-time RT-PCR and chemotaxis assays were performed at least three times in duplicate. *p < 0.01, **p < 0.001.
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Figure 8: MET receptor downregulation decreases CXCR4 expression and function(A) Immunohistochemical analysis of CXCR4 receptor expression in human tissue. (B) Real-time RT-PCR and flow cytometry analysis revealed significant decreases in MET transcript and protein levels in MET receptor-silenced cells (shMET) relative to controls (wild type, WT, and shLacZ). (C) Western blot analysis of CXCR4 activation. AKT and MAPK phosphorylation was tested after after HGF and SDF-1 stimulation. (D) In chemotaxis assay MET-deficient CC cells showed limited chemotactic activity towards HGF and/or SDF-1 gradient. Western blot and FACS analyses were performed three times with similar results; representative results are shown. Real-time RT-PCR and chemotaxis assays were performed at least three times in duplicate. *p < 0.01, **p < 0.001.

Mentions: SDF-1-CXCR4 axis is a key player responsible for invasive phenotype and metastatic behavior of many tumor types [30, 31]. In our study, we evaluated the level of CXCR4 expression in human samples (n = 31) obtained from cervical carcinoma patients. Regardless of the cancer stage, we observed strong and very strong expression of CXCR4 receptor (Figure 8A; Supplementary Table 4). We became interested whether downregulation of MET receptor influences SDF-1/CXCR4 axis. We noticed that the expression of CXCR4 was strongly decreased in shMET HTB-35 and unchanged in control HTB-35 cell lines (Figure 8B). After the activation of CXCR4 receptor with SDF-1 we found that control cells responded to the ligand by phosphorylation of AKT and MAPK kinases. However, the activation of MAPK and AKT after SDF-1 stimulation was highly impaired in shMET HTB-35 cells (Figure 8C). These data led us to the question how MET receptor influence the chemotactive activity of CXCR4 receptor. We found that downregulation of MET receptor drastically impaired directed migration towards HGF and SDF-1 ligands (Figure 8D). HTB-34 control and shMET cells did not express CXCR4 receptor (Supplementary Figure 2).


MET receptor is a potential therapeutic target in high grade cervical cancer.

Miekus K, Pawlowska M, Sekuła M, Drabik G, Madeja Z, Adamek D, Majka M - Oncotarget (2015)

MET receptor downregulation decreases CXCR4 expression and function(A) Immunohistochemical analysis of CXCR4 receptor expression in human tissue. (B) Real-time RT-PCR and flow cytometry analysis revealed significant decreases in MET transcript and protein levels in MET receptor-silenced cells (shMET) relative to controls (wild type, WT, and shLacZ). (C) Western blot analysis of CXCR4 activation. AKT and MAPK phosphorylation was tested after after HGF and SDF-1 stimulation. (D) In chemotaxis assay MET-deficient CC cells showed limited chemotactic activity towards HGF and/or SDF-1 gradient. Western blot and FACS analyses were performed three times with similar results; representative results are shown. Real-time RT-PCR and chemotaxis assays were performed at least three times in duplicate. *p < 0.01, **p < 0.001.
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Figure 8: MET receptor downregulation decreases CXCR4 expression and function(A) Immunohistochemical analysis of CXCR4 receptor expression in human tissue. (B) Real-time RT-PCR and flow cytometry analysis revealed significant decreases in MET transcript and protein levels in MET receptor-silenced cells (shMET) relative to controls (wild type, WT, and shLacZ). (C) Western blot analysis of CXCR4 activation. AKT and MAPK phosphorylation was tested after after HGF and SDF-1 stimulation. (D) In chemotaxis assay MET-deficient CC cells showed limited chemotactic activity towards HGF and/or SDF-1 gradient. Western blot and FACS analyses were performed three times with similar results; representative results are shown. Real-time RT-PCR and chemotaxis assays were performed at least three times in duplicate. *p < 0.01, **p < 0.001.
Mentions: SDF-1-CXCR4 axis is a key player responsible for invasive phenotype and metastatic behavior of many tumor types [30, 31]. In our study, we evaluated the level of CXCR4 expression in human samples (n = 31) obtained from cervical carcinoma patients. Regardless of the cancer stage, we observed strong and very strong expression of CXCR4 receptor (Figure 8A; Supplementary Table 4). We became interested whether downregulation of MET receptor influences SDF-1/CXCR4 axis. We noticed that the expression of CXCR4 was strongly decreased in shMET HTB-35 and unchanged in control HTB-35 cell lines (Figure 8B). After the activation of CXCR4 receptor with SDF-1 we found that control cells responded to the ligand by phosphorylation of AKT and MAPK kinases. However, the activation of MAPK and AKT after SDF-1 stimulation was highly impaired in shMET HTB-35 cells (Figure 8C). These data led us to the question how MET receptor influence the chemotactive activity of CXCR4 receptor. We found that downregulation of MET receptor drastically impaired directed migration towards HGF and SDF-1 ligands (Figure 8D). HTB-34 control and shMET cells did not express CXCR4 receptor (Supplementary Figure 2).

Bottom Line: MET receptor downregulation also resulted in decreased cyclin D1 and c-myc levels but did not increase apoptosis.Subsequent experiments showed that downregulation of the MET receptor decreased the expression of a key regulator of the epithelial-to-mesenchymal transition, SLUG. and increased the expression of E-cadherin, a hallmark of the epithelial phenotype.Taken together, our results strongly suggest that the MET receptor influences the oncogenic properties of cervical carcinoma cells in vitro and in vivo.

View Article: PubMed Central - PubMed

Affiliation: Department of General Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Cracow, Poland.

ABSTRACT
Cervical cancer is one of the leading causes of death among women suffering from tumors. Current treatment options are insufficient. Here, we investigated the MET receptor as a potential molecular target in advanced cervical cancer. Downregulation of MET receptor expression via RNA interference in different cervical carcinoma cell lines dramatically decreased tumor growth and forced tumor differentiation in vivo. MET receptor silencing also led to a dramatic decrease in cell size and a decrease in proliferation rate under normal and stress conditions. MET receptor downregulation also resulted in decreased cyclin D1 and c-myc levels but did not increase apoptosis. Subsequent experiments showed that downregulation of the MET receptor decreased the expression of a key regulator of the epithelial-to-mesenchymal transition, SLUG. and increased the expression of E-cadherin, a hallmark of the epithelial phenotype. Moreover, MET downregulation impairs expression and signaling of CXCR4 receptor, responsible for invasive phenotype. Taken together, our results strongly suggest that the MET receptor influences the oncogenic properties of cervical carcinoma cells in vitro and in vivo. These findings highlight a unique role of the MET receptor in cervical carcinoma cells and indicate the MET receptor as a potential therapeutic target for advanced cervical carcinoma.

No MeSH data available.


Related in: MedlinePlus