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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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Expression of GALR1 and GALR2 using individual recombinant AAV vectors in head and neck squamous cell carcinoma (HNSCC) cells. (a) Quantitative RT-PCR was performed to measure GALR1 and GALR2 expression in wild type (Wt) or HNSCC cells transduced by each of the rAAV vectors. (b) Western blot results showing exogenous GALR1 and GALR2 expression in Wt or HNSCC cells transduced by each of the rAAV vectors, as detected by an HA-antibody.
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fig02: Expression of GALR1 and GALR2 using individual recombinant AAV vectors in head and neck squamous cell carcinoma (HNSCC) cells. (a) Quantitative RT-PCR was performed to measure GALR1 and GALR2 expression in wild type (Wt) or HNSCC cells transduced by each of the rAAV vectors. (b) Western blot results showing exogenous GALR1 and GALR2 expression in Wt or HNSCC cells transduced by each of the rAAV vectors, as detected by an HA-antibody.

Mentions: We determined both the endogenous and exogenous mRNA expression levels of each GALR gene in HEp-2 and KB cells transduced by rAAV-GALR1 or rAAV-GALR2 by quantitative RT-PCR. After 48 h of transduction, elevated mRNA expression levels of exogenous GALR1 and GALR2 were observed in both cell lines (Fig.2a).


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)

Expression of GALR1 and GALR2 using individual recombinant AAV vectors in head and neck squamous cell carcinoma (HNSCC) cells. (a) Quantitative RT-PCR was performed to measure GALR1 and GALR2 expression in wild type (Wt) or HNSCC cells transduced by each of the rAAV vectors. (b) Western blot results showing exogenous GALR1 and GALR2 expression in Wt or HNSCC cells transduced by each of the rAAV vectors, as detected by an HA-antibody.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig02: Expression of GALR1 and GALR2 using individual recombinant AAV vectors in head and neck squamous cell carcinoma (HNSCC) cells. (a) Quantitative RT-PCR was performed to measure GALR1 and GALR2 expression in wild type (Wt) or HNSCC cells transduced by each of the rAAV vectors. (b) Western blot results showing exogenous GALR1 and GALR2 expression in Wt or HNSCC cells transduced by each of the rAAV vectors, as detected by an HA-antibody.
Mentions: We determined both the endogenous and exogenous mRNA expression levels of each GALR gene in HEp-2 and KB cells transduced by rAAV-GALR1 or rAAV-GALR2 by quantitative RT-PCR. After 48 h of transduction, elevated mRNA expression levels of exogenous GALR1 and GALR2 were observed in both cell lines (Fig.2a).

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