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

HSVtk/GCV therapy using the GSDMB enhancer-driven lentivirus vector improved the survival rate of PD mice. (a) A lentiviral therapeutic vector for GSDMB enhancer (Enh)-driven expression of herpes simplex virus thymidine kinase (HSVtk). (b) Cell proliferation assays on 60As6 and Met-5A transduced with the therapeutic vector, performed by incubation in the medium with (+)/without (−) ganciclovir (GCV). (c) A regimen of HSVtk/GCV therapy for PD mice. Bar, standard deviation, P, P -value of Student’s t-test between the cultured cells with (+) and without (−) GCV. (d) Microscopic observation exhibited a small population of 60As6GFP cells (green fluorescence) implanted into mouse peritoneum at day 10. (e) Number of survived mice after HSVtk/GCV therapy with the sense-strand expressing vector (red) and with an antisense-strand expressing vector as reference (blue). Mean survival time of each group is shown at the right side with P- value of Student’s t-test between the two groups
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Fig4: HSVtk/GCV therapy using the GSDMB enhancer-driven lentivirus vector improved the survival rate of PD mice. (a) A lentiviral therapeutic vector for GSDMB enhancer (Enh)-driven expression of herpes simplex virus thymidine kinase (HSVtk). (b) Cell proliferation assays on 60As6 and Met-5A transduced with the therapeutic vector, performed by incubation in the medium with (+)/without (−) ganciclovir (GCV). (c) A regimen of HSVtk/GCV therapy for PD mice. Bar, standard deviation, P, P -value of Student’s t-test between the cultured cells with (+) and without (−) GCV. (d) Microscopic observation exhibited a small population of 60As6GFP cells (green fluorescence) implanted into mouse peritoneum at day 10. (e) Number of survived mice after HSVtk/GCV therapy with the sense-strand expressing vector (red) and with an antisense-strand expressing vector as reference (blue). Mean survival time of each group is shown at the right side with P- value of Student’s t-test between the two groups

Mentions: Next, we examined the effect of the HSVtk/GCV therapy using the GSDMB enhancer-driven HSVtk lentivirus vector on 60As6 in vitro (Fig. 4a). The number of 60As6 cells transduced with the lentivirus vector was significantly reduced when incubated in medium supplemented with GCV; on the other hand, the same HSVtk/GCV treatment had no effect on the cell number of Met-5A (Fig. 4b).Fig. 4


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)

HSVtk/GCV therapy using the GSDMB enhancer-driven lentivirus vector improved the survival rate of PD mice. (a) A lentiviral therapeutic vector for GSDMB enhancer (Enh)-driven expression of herpes simplex virus thymidine kinase (HSVtk). (b) Cell proliferation assays on 60As6 and Met-5A transduced with the therapeutic vector, performed by incubation in the medium with (+)/without (−) ganciclovir (GCV). (c) A regimen of HSVtk/GCV therapy for PD mice. Bar, standard deviation, P, P -value of Student’s t-test between the cultured cells with (+) and without (−) GCV. (d) Microscopic observation exhibited a small population of 60As6GFP cells (green fluorescence) implanted into mouse peritoneum at day 10. (e) Number of survived mice after HSVtk/GCV therapy with the sense-strand expressing vector (red) and with an antisense-strand expressing vector as reference (blue). Mean survival time of each group is shown at the right side with P- value of Student’s t-test between the two groups
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Fig4: HSVtk/GCV therapy using the GSDMB enhancer-driven lentivirus vector improved the survival rate of PD mice. (a) A lentiviral therapeutic vector for GSDMB enhancer (Enh)-driven expression of herpes simplex virus thymidine kinase (HSVtk). (b) Cell proliferation assays on 60As6 and Met-5A transduced with the therapeutic vector, performed by incubation in the medium with (+)/without (−) ganciclovir (GCV). (c) A regimen of HSVtk/GCV therapy for PD mice. Bar, standard deviation, P, P -value of Student’s t-test between the cultured cells with (+) and without (−) GCV. (d) Microscopic observation exhibited a small population of 60As6GFP cells (green fluorescence) implanted into mouse peritoneum at day 10. (e) Number of survived mice after HSVtk/GCV therapy with the sense-strand expressing vector (red) and with an antisense-strand expressing vector as reference (blue). Mean survival time of each group is shown at the right side with P- value of Student’s t-test between the two groups
Mentions: Next, we examined the effect of the HSVtk/GCV therapy using the GSDMB enhancer-driven HSVtk lentivirus vector on 60As6 in vitro (Fig. 4a). The number of 60As6 cells transduced with the lentivirus vector was significantly reduced when incubated in medium supplemented with GCV; on the other hand, the same HSVtk/GCV treatment had no effect on the cell number of Met-5A (Fig. 4b).Fig. 4

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