Limits...
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.

Show MeSH

Related in: MedlinePlus

Transduction efficiency using recombinant adeno-associated virus (rAAV)-enhanced green fluorescent protein (EGFP) in head and neck squamous cell carcinoma (HNSCC) cell lines. HEp-2 and KB cells were transduced by AAV-EGFP at various vector doses. Upper: GFP-positive cell rate under vector dose-dependent and time-course conditions in each cell line are indicated. Lower: Raw data expressing GFP intensity as a measure of gene expression.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4317884&req=5

fig01: Transduction efficiency using recombinant adeno-associated virus (rAAV)-enhanced green fluorescent protein (EGFP) in head and neck squamous cell carcinoma (HNSCC) cell lines. HEp-2 and KB cells were transduced by AAV-EGFP at various vector doses. Upper: GFP-positive cell rate under vector dose-dependent and time-course conditions in each cell line are indicated. Lower: Raw data expressing GFP intensity as a measure of gene expression.

Mentions: To determine transduction efficiency in HNSCC cell lines using rAAV vectors, HEp-2 and KB cells were infected with a rAAV-EGFP vector at various vector doses. After incubation for 24–72 h, GFP-positive cells were counted by flow cytometry. We confirmed a high GFP expression (>90%) in both HNSCC cell lines transduced by vector doses above 1 × 105 vector genomes (vg)/cell for 48 h (Fig.1). Consequently, a titer of 1 × 105 vg/cell was used as standard vector dose.


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)

Transduction efficiency using recombinant adeno-associated virus (rAAV)-enhanced green fluorescent protein (EGFP) in head and neck squamous cell carcinoma (HNSCC) cell lines. HEp-2 and KB cells were transduced by AAV-EGFP at various vector doses. Upper: GFP-positive cell rate under vector dose-dependent and time-course conditions in each cell line are indicated. Lower: Raw data expressing GFP intensity as a measure of gene expression.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig01: Transduction efficiency using recombinant adeno-associated virus (rAAV)-enhanced green fluorescent protein (EGFP) in head and neck squamous cell carcinoma (HNSCC) cell lines. HEp-2 and KB cells were transduced by AAV-EGFP at various vector doses. Upper: GFP-positive cell rate under vector dose-dependent and time-course conditions in each cell line are indicated. Lower: Raw data expressing GFP intensity as a measure of gene expression.
Mentions: To determine transduction efficiency in HNSCC cell lines using rAAV vectors, HEp-2 and KB cells were infected with a rAAV-EGFP vector at various vector doses. After incubation for 24–72 h, GFP-positive cells were counted by flow cytometry. We confirmed a high GFP expression (>90%) in both HNSCC cell lines transduced by vector doses above 1 × 105 vector genomes (vg)/cell for 48 h (Fig.1). Consequently, a titer of 1 × 105 vg/cell was used as standard vector dose.

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