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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 downregulation alters proliferation/viability under stress conditionsMET receptor downregulation via a lentiviral vector containing anti-MET shRNA resulted in decreases at mRNA (A) and protein (B, C) levels. Downregulation of MET receptor alters proliferation/viability under hypoxia (D) and starvation (E) conditions. A – Real-time RT-PCR revealed significant decreases in MET transcript levels in MET receptor-silenced cells (shMET) relative to controls (wild type, WT, and shLacZ) in all tested cell lines (HTB-34, HeLa and HTB-35). B – Flow cytometry analysis of MET receptor expression. C – Western blot analysis revealed complete downregulation of MET receptor expression in shMET HTB-34, HeLa and HTB-35 cells. D. MTT assay of cells cultured under starvation conditions (MEM supplemented with 0.5% BSA). E – MTT assay of cells cultured under hypoxic conditions (2% oxygen). Western blot and FACS analyses were performed at least three times with similar results; representative results are shown. Real-time RT-PCR was performed at least three times in duplicates. MTT assay was repeated three times in triplicates. *p < 0.01, **p < 0.001.
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Figure 2: MET downregulation alters proliferation/viability under stress conditionsMET receptor downregulation via a lentiviral vector containing anti-MET shRNA resulted in decreases at mRNA (A) and protein (B, C) levels. Downregulation of MET receptor alters proliferation/viability under hypoxia (D) and starvation (E) conditions. A – Real-time RT-PCR revealed significant decreases in MET transcript levels in MET receptor-silenced cells (shMET) relative to controls (wild type, WT, and shLacZ) in all tested cell lines (HTB-34, HeLa and HTB-35). B – Flow cytometry analysis of MET receptor expression. C – Western blot analysis revealed complete downregulation of MET receptor expression in shMET HTB-34, HeLa and HTB-35 cells. D. MTT assay of cells cultured under starvation conditions (MEM supplemented with 0.5% BSA). E – MTT assay of cells cultured under hypoxic conditions (2% oxygen). Western blot and FACS analyses were performed at least three times with similar results; representative results are shown. Real-time RT-PCR was performed at least three times in duplicates. MTT assay was repeated three times in triplicates. *p < 0.01, **p < 0.001.

Mentions: Cervical carcinoma cells were transduced with lentiviral vectors containing anti-MET shRNAs that were established in our laboratory [23]. The efficiency of MET downregulation was assessed in cells transduced with control LacZ (shLacZ) and MET (shMET) shRNA and compared with control wild-type (WT) cells. MET receptor expression levels were evaluated at the mRNA level using real-time RT–PCR (Figure 2A) and at the protein level using flow cytometry (Figure 2B) and western blot analysis (Figure 2C). The functionality of the silenced receptor was tested by a chemotaxis assay (Supplementary Figure 1).


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 downregulation alters proliferation/viability under stress conditionsMET receptor downregulation via a lentiviral vector containing anti-MET shRNA resulted in decreases at mRNA (A) and protein (B, C) levels. Downregulation of MET receptor alters proliferation/viability under hypoxia (D) and starvation (E) conditions. A – Real-time RT-PCR revealed significant decreases in MET transcript levels in MET receptor-silenced cells (shMET) relative to controls (wild type, WT, and shLacZ) in all tested cell lines (HTB-34, HeLa and HTB-35). B – Flow cytometry analysis of MET receptor expression. C – Western blot analysis revealed complete downregulation of MET receptor expression in shMET HTB-34, HeLa and HTB-35 cells. D. MTT assay of cells cultured under starvation conditions (MEM supplemented with 0.5% BSA). E – MTT assay of cells cultured under hypoxic conditions (2% oxygen). Western blot and FACS analyses were performed at least three times with similar results; representative results are shown. Real-time RT-PCR was performed at least three times in duplicates. MTT assay was repeated three times in triplicates. *p < 0.01, **p < 0.001.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4496342&req=5

Figure 2: MET downregulation alters proliferation/viability under stress conditionsMET receptor downregulation via a lentiviral vector containing anti-MET shRNA resulted in decreases at mRNA (A) and protein (B, C) levels. Downregulation of MET receptor alters proliferation/viability under hypoxia (D) and starvation (E) conditions. A – Real-time RT-PCR revealed significant decreases in MET transcript levels in MET receptor-silenced cells (shMET) relative to controls (wild type, WT, and shLacZ) in all tested cell lines (HTB-34, HeLa and HTB-35). B – Flow cytometry analysis of MET receptor expression. C – Western blot analysis revealed complete downregulation of MET receptor expression in shMET HTB-34, HeLa and HTB-35 cells. D. MTT assay of cells cultured under starvation conditions (MEM supplemented with 0.5% BSA). E – MTT assay of cells cultured under hypoxic conditions (2% oxygen). Western blot and FACS analyses were performed at least three times with similar results; representative results are shown. Real-time RT-PCR was performed at least three times in duplicates. MTT assay was repeated three times in triplicates. *p < 0.01, **p < 0.001.
Mentions: Cervical carcinoma cells were transduced with lentiviral vectors containing anti-MET shRNAs that were established in our laboratory [23]. The efficiency of MET downregulation was assessed in cells transduced with control LacZ (shLacZ) and MET (shMET) shRNA and compared with control wild-type (WT) cells. MET receptor expression levels were evaluated at the mRNA level using real-time RT–PCR (Figure 2A) and at the protein level using flow cytometry (Figure 2B) and western blot analysis (Figure 2C). The functionality of the silenced receptor was tested by a chemotaxis assay (Supplementary Figure 1).

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