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Novel anti-tumor mechanism of galanin receptor type 2 in head and neck squamous cell carcinoma cells.

Uehara T, Kanazawa T, Mizukami H, Uchibori R, Tsukahara T, Urabe M, Kume A, Misawa K, Carey TE, Suzuki M, Ichimura K, Ozawa K - Cancer Sci. (2013)

Bottom Line: In this study, we first transduced HEp-2 and KB cell lines using a recombinant adeno-associated virus (rAAV)-green fluorescent protein (GFP) vector and confirmed a high GFP expression rate (>90%) in both cell lines at the standard vector dose.Additionally, the annexin V-positive rate and sub-G0/G1 phase population were significantly elevated in HEp-2 cells (mock vs GALR2: 12.3 vs 25.0% (P < 0.01) and 9.1 vs 32.0% (P < 0.05), respectively) after 48 h.Furthermore, in HEp-2 cells, GALR2-mediated apoptosis was caspase-independent, involving downregulation of ERK1/2, followed by induction of the pro-apoptotic Bcl-2 protein, Bim.

View Article: PubMed Central - PubMed

Affiliation: Department of Otorhinolaryngology, Head and Neck Surgery, Graduate School of Medicine, University of the Ryukyus, Nishihara, Japan; Division of Genetic Therapeutics, Center for Molecular Medicine, Jichi Medical University, Shimotsuke, Japan; Department of Otolaryngology, Head and Neck Surgery, Jichi Medical University School of Medicine, Shimotsuke, Japan.

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Immunoblotting analysis of the phosphorylation of ERK1/2 and Akt and regulation of key apoptosis regulators by co-administration of recombinant adeno-associated virus (rAAV)-GALR2 vector and galanin. (a) Effect of galanin on ERK1/2 activation and Bim expression in rAAV-GALR2 vector-transduced head and neck squamous cell carcinoma (HNSCC) cells. (b) Effect of galanin on Akt activation in GALR-transduced HNSCC cells. (c) Effects of treatment of cells with galanin and transduction with individual rAAV vectors on the phosphorylation state of ERK1/2 and expression of proteins belonging to the Bcl-2 or IAP families.
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fig05: Immunoblotting analysis of the phosphorylation of ERK1/2 and Akt and regulation of key apoptosis regulators by co-administration of recombinant adeno-associated virus (rAAV)-GALR2 vector and galanin. (a) Effect of galanin on ERK1/2 activation and Bim expression in rAAV-GALR2 vector-transduced head and neck squamous cell carcinoma (HNSCC) cells. (b) Effect of galanin on Akt activation in GALR-transduced HNSCC cells. (c) Effects of treatment of cells with galanin and transduction with individual rAAV vectors on the phosphorylation state of ERK1/2 and expression of proteins belonging to the Bcl-2 or IAP families.

Mentions: As the GALR2-mediated cytotoxic effects were mainly due to apoptosis induction, we examined whether stimulation of the GALR2 signaling pathway affected the phosphorylation states of ERK1/2 and Akt by immunoblotting. Sustained dephosphorylation of ERK1/2 was induced by stimulation of GALR2 signaling in both HNSCC cell lines (Fig.5a), but no effect on Akt phosphorylation was observed (Fig.5b).


Novel anti-tumor mechanism of galanin receptor type 2 in head and neck squamous cell carcinoma cells.

Uehara T, Kanazawa T, Mizukami H, Uchibori R, Tsukahara T, Urabe M, Kume A, Misawa K, Carey TE, Suzuki M, Ichimura K, Ozawa K - Cancer Sci. (2013)

Immunoblotting analysis of the phosphorylation of ERK1/2 and Akt and regulation of key apoptosis regulators by co-administration of recombinant adeno-associated virus (rAAV)-GALR2 vector and galanin. (a) Effect of galanin on ERK1/2 activation and Bim expression in rAAV-GALR2 vector-transduced head and neck squamous cell carcinoma (HNSCC) cells. (b) Effect of galanin on Akt activation in GALR-transduced HNSCC cells. (c) Effects of treatment of cells with galanin and transduction with individual rAAV vectors on the phosphorylation state of ERK1/2 and expression of proteins belonging to the Bcl-2 or IAP families.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4317884&req=5

fig05: Immunoblotting analysis of the phosphorylation of ERK1/2 and Akt and regulation of key apoptosis regulators by co-administration of recombinant adeno-associated virus (rAAV)-GALR2 vector and galanin. (a) Effect of galanin on ERK1/2 activation and Bim expression in rAAV-GALR2 vector-transduced head and neck squamous cell carcinoma (HNSCC) cells. (b) Effect of galanin on Akt activation in GALR-transduced HNSCC cells. (c) Effects of treatment of cells with galanin and transduction with individual rAAV vectors on the phosphorylation state of ERK1/2 and expression of proteins belonging to the Bcl-2 or IAP families.
Mentions: As the GALR2-mediated cytotoxic effects were mainly due to apoptosis induction, we examined whether stimulation of the GALR2 signaling pathway affected the phosphorylation states of ERK1/2 and Akt by immunoblotting. Sustained dephosphorylation of ERK1/2 was induced by stimulation of GALR2 signaling in both HNSCC cell lines (Fig.5a), but no effect on Akt phosphorylation was observed (Fig.5b).

Bottom Line: In this study, we first transduced HEp-2 and KB cell lines using a recombinant adeno-associated virus (rAAV)-green fluorescent protein (GFP) vector and confirmed a high GFP expression rate (>90%) in both cell lines at the standard vector dose.Additionally, the annexin V-positive rate and sub-G0/G1 phase population were significantly elevated in HEp-2 cells (mock vs GALR2: 12.3 vs 25.0% (P < 0.01) and 9.1 vs 32.0% (P < 0.05), respectively) after 48 h.Furthermore, in HEp-2 cells, GALR2-mediated apoptosis was caspase-independent, involving downregulation of ERK1/2, followed by induction of the pro-apoptotic Bcl-2 protein, Bim.

View Article: PubMed Central - PubMed

Affiliation: Department of Otorhinolaryngology, Head and Neck Surgery, Graduate School of Medicine, University of the Ryukyus, Nishihara, Japan; Division of Genetic Therapeutics, Center for Molecular Medicine, Jichi Medical University, Shimotsuke, Japan; Department of Otolaryngology, Head and Neck Surgery, Jichi Medical University School of Medicine, Shimotsuke, Japan.

Show MeSH
Related in: MedlinePlus