Limits...
A GSDMB enhancer-driven HSV thymidine kinase-expressing vector for controlling occult peritoneal dissemination of gastric cancer cells.

Saeki N, Komatsuzaki R, Chiwaki F, Yanagihara K, Sasaki H - BMC Cancer (2015)

Bottom Line: The lentiviral therapeutic vector suppressed proliferation of a GC cell line, 60As6, in vitro in the presence of ganciclovir, and intraperitoneal administration of the vector prolonged the survival term of mice that were intraperitoneally inoculated with 60As6 one week prior to the administration.The GSDMB-driven HSVtk expression vector had a therapeutic effect on the occult PD model mice.This strategy can potentially be used to treat GC patients with PD.

View Article: PubMed Central - PubMed

Affiliation: Division of Genetics, National Cancer Center Research Institute, Tsukiji 5-1-1, Chuo-ku, Tokyo, 104-0045, Japan. nsaeki@ncc.go.jp.

ABSTRACT

Background: Gastric cancer (GC) is one of the major malignant diseases worldwide, especially in Asia, and Japan and Korea have the highest incidence in the world. Because most of the cases that are refractory to therapies die due to peritoneal dissemination (PD) of the cancer cells, controlling PD is important for patient survival. GSDMB is a member of the gasdermin gene family. Because GSDMB is expressed in many types of cancer, including GC, it is likely that the gene contains a regulatory region that is utilized for therapy of occult PD through cancer cell-specific expression of cytotoxic genes.

Methods: We performed reporter assays to identify the regulatory region for the cancer cell-specific expression. We also constructed a lentiviral therapeutic vector that expresses herpes simplex virus thymidine kinase (HSVtk) in a GC cell-specific manner, and tested it in a mouse model of PD.

Results: We identified the regulatory region at +496 to +989 from the GSDMB transcription start site and designated it as a GSDMB enhancer. The lentiviral therapeutic vector suppressed proliferation of a GC cell line, 60As6, in vitro in the presence of ganciclovir, and intraperitoneal administration of the vector prolonged the survival term of mice that were intraperitoneally inoculated with 60As6 one week prior to the administration.

Conclusions: The GSDMB-driven HSVtk expression vector had a therapeutic effect on the occult PD model mice. This strategy can potentially be used to treat GC patients with PD.

No MeSH data available.


Related in: MedlinePlus

GSDMB enhancer has strong transcriptional activity in a 60As6 cell line. Luciferase assays with three types of cultured cells: 60As6 cells that were used for making peritoneal dissemination (PD) model mice in this study, primary culture cells of mouse peritoneal mesothelial cells and established human mesotherial cell line Met-5A. Bar, standard deviation
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC4446855&req=5

Fig3: GSDMB enhancer has strong transcriptional activity in a 60As6 cell line. Luciferase assays with three types of cultured cells: 60As6 cells that were used for making peritoneal dissemination (PD) model mice in this study, primary culture cells of mouse peritoneal mesothelial cells and established human mesotherial cell line Met-5A. Bar, standard deviation

Mentions: We previously reported a mouse PD model (PD mice) that was produced by intraperitoneal injection of 60As6 cells [25]; in this study, we developed a viral therapeutic vector for the treatment of PD mice. For examining the strength of the transcriptional activity of GSDMB enhancer in 60As6, reporter assays were performed, using the reporter construct for the upstream regions of CXCR4 and CXCR7 for comparison. The GSDMB enhancer showed stronger transcriptional activity in 60As6 cells than the CXCR4 or the CXCR7 upstream regions, and, importantly, the GSDMB enhancer had very weak transcriptional activity in mouse peritoneal mesothelial cells and in Met-5A, a human mesothelial cell line (Fig. 3). This result suggests that the GSDMB enhancer enables HSVtk expression almost exclusively in 60As6 but not in mesothelial cells of the peritoneal cavity of the PD mice, and probably not in human peritoneal mesothelium.Fig. 3


A GSDMB enhancer-driven HSV thymidine kinase-expressing vector for controlling occult peritoneal dissemination of gastric cancer cells.

Saeki N, Komatsuzaki R, Chiwaki F, Yanagihara K, Sasaki H - BMC Cancer (2015)

GSDMB enhancer has strong transcriptional activity in a 60As6 cell line. Luciferase assays with three types of cultured cells: 60As6 cells that were used for making peritoneal dissemination (PD) model mice in this study, primary culture cells of mouse peritoneal mesothelial cells and established human mesotherial cell line Met-5A. Bar, standard deviation
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4446855&req=5

Fig3: GSDMB enhancer has strong transcriptional activity in a 60As6 cell line. Luciferase assays with three types of cultured cells: 60As6 cells that were used for making peritoneal dissemination (PD) model mice in this study, primary culture cells of mouse peritoneal mesothelial cells and established human mesotherial cell line Met-5A. Bar, standard deviation
Mentions: We previously reported a mouse PD model (PD mice) that was produced by intraperitoneal injection of 60As6 cells [25]; in this study, we developed a viral therapeutic vector for the treatment of PD mice. For examining the strength of the transcriptional activity of GSDMB enhancer in 60As6, reporter assays were performed, using the reporter construct for the upstream regions of CXCR4 and CXCR7 for comparison. The GSDMB enhancer showed stronger transcriptional activity in 60As6 cells than the CXCR4 or the CXCR7 upstream regions, and, importantly, the GSDMB enhancer had very weak transcriptional activity in mouse peritoneal mesothelial cells and in Met-5A, a human mesothelial cell line (Fig. 3). This result suggests that the GSDMB enhancer enables HSVtk expression almost exclusively in 60As6 but not in mesothelial cells of the peritoneal cavity of the PD mice, and probably not in human peritoneal mesothelium.Fig. 3

Bottom Line: The lentiviral therapeutic vector suppressed proliferation of a GC cell line, 60As6, in vitro in the presence of ganciclovir, and intraperitoneal administration of the vector prolonged the survival term of mice that were intraperitoneally inoculated with 60As6 one week prior to the administration.The GSDMB-driven HSVtk expression vector had a therapeutic effect on the occult PD model mice.This strategy can potentially be used to treat GC patients with PD.

View Article: PubMed Central - PubMed

Affiliation: Division of Genetics, National Cancer Center Research Institute, Tsukiji 5-1-1, Chuo-ku, Tokyo, 104-0045, Japan. nsaeki@ncc.go.jp.

ABSTRACT

Background: Gastric cancer (GC) is one of the major malignant diseases worldwide, especially in Asia, and Japan and Korea have the highest incidence in the world. Because most of the cases that are refractory to therapies die due to peritoneal dissemination (PD) of the cancer cells, controlling PD is important for patient survival. GSDMB is a member of the gasdermin gene family. Because GSDMB is expressed in many types of cancer, including GC, it is likely that the gene contains a regulatory region that is utilized for therapy of occult PD through cancer cell-specific expression of cytotoxic genes.

Methods: We performed reporter assays to identify the regulatory region for the cancer cell-specific expression. We also constructed a lentiviral therapeutic vector that expresses herpes simplex virus thymidine kinase (HSVtk) in a GC cell-specific manner, and tested it in a mouse model of PD.

Results: We identified the regulatory region at +496 to +989 from the GSDMB transcription start site and designated it as a GSDMB enhancer. The lentiviral therapeutic vector suppressed proliferation of a GC cell line, 60As6, in vitro in the presence of ganciclovir, and intraperitoneal administration of the vector prolonged the survival term of mice that were intraperitoneally inoculated with 60As6 one week prior to the administration.

Conclusions: The GSDMB-driven HSVtk expression vector had a therapeutic effect on the occult PD model mice. This strategy can potentially be used to treat GC patients with PD.

No MeSH data available.


Related in: MedlinePlus