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Recombinant adenovirus IL-24-Bax promotes apoptosis of hepatocellular carcinoma cells in vitro and in vivo.

Li J, Shi L, Zhang X, Kang X, Wen Y, Qian H, Zhou Y, Xu W, Zhang Y, Wu M, Yin Z - Cancer Gene Ther. (2010)

Bottom Line: The mechanism of this response may include the effect of the 10HRE/VEGF385 promoter and the synergistic effect of IL-24 and Bax.Ad.IL-24-Bax also suppressed tumor growth in nude mice and induced apoptosis.Ad.IL-24-Bax may be a useful tool for gene therapy of hepatic cancer.

View Article: PubMed Central - PubMed

Affiliation: Molecular Oncology Laboratory, Eastern Hepatobiliary Surgery Hospital, Second Military Medical University, Shanghai, China.

ABSTRACT
Gene therapy promises to become an alternative choice for the treatment of hepatic cancer. In many cancers, the delivery of chimeric proteins by adenovirus vector has been reported to induce apoptosis. This study was performed to evaluate whether the recombinant adenovirus interleukin (IL)-24-Bax can induce apoptosis in hepatocellular carcinoma cells in vitro and in vivo. Several recombinant adenoviruses were constructed, and the expression of their encoded proteins was measured. The effects of the recombinant adenovirus on hepatocellular carcinoma cells and the normal hepatocyte cell line were investigated through cell viability and apoptosis assays after the cells were treated with Ad.Luc, Ad.IL-24, Ad.Bax or Ad.IL-24-Bax. The mechanism involved was also explored. A tumor-bearing mouse model was used to evaluate the effects of the adenovirus on tumor volume and cell apoptosis in vivo. Ad.IL-24-Bax selectively suppressed growth of hepatocellular carcinoma cells and induced apoptosis, but it had little influence on the normal hepatocytes. The mechanism of this response may include the effect of the 10HRE/VEGF385 promoter and the synergistic effect of IL-24 and Bax. Ad.IL-24-Bax also suppressed tumor growth in nude mice and induced apoptosis. Ad.IL-24-Bax may be a useful tool for gene therapy of hepatic cancer.

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Related in: MedlinePlus

Construction and expression of Ad.IL-24-Bax. (a) Construction of recombinant adenovirus vector that encodes the IL-24-Bax. (b) Western blot analysis of the expression of the IL-24-Bax fusion protein. Hep3B cells were treated with Ad.IL-24-Bax, Ad.IL-24, Ad.Bax or Ad.Luc. Forty-eight hours after treatment, proteins were extracted from cells for western blot analysis. β-Actin was used as the internal standard.
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fig1: Construction and expression of Ad.IL-24-Bax. (a) Construction of recombinant adenovirus vector that encodes the IL-24-Bax. (b) Western blot analysis of the expression of the IL-24-Bax fusion protein. Hep3B cells were treated with Ad.IL-24-Bax, Ad.IL-24, Ad.Bax or Ad.Luc. Forty-eight hours after treatment, proteins were extracted from cells for western blot analysis. β-Actin was used as the internal standard.

Mentions: As Figure 1a shows, the recombinant adenovirus IL-24-Bax was constructed as well as Ad.IL-24, Ad.Bax and Ad.Luc. When Hep3B cells were transfected with Ad.IL-24-Bax, the transfection efficiency was related to the MOI of the recombinant adenovirus. As the MOI increased, the efficiency of transfection also increased. When MOI was higher than 25, the efficiency of transfection was as high as 90%. Considering the cytotoxicity of the virus, an MOI of 10 for the virus was used in transfection experiments in this study.


Recombinant adenovirus IL-24-Bax promotes apoptosis of hepatocellular carcinoma cells in vitro and in vivo.

Li J, Shi L, Zhang X, Kang X, Wen Y, Qian H, Zhou Y, Xu W, Zhang Y, Wu M, Yin Z - Cancer Gene Ther. (2010)

Construction and expression of Ad.IL-24-Bax. (a) Construction of recombinant adenovirus vector that encodes the IL-24-Bax. (b) Western blot analysis of the expression of the IL-24-Bax fusion protein. Hep3B cells were treated with Ad.IL-24-Bax, Ad.IL-24, Ad.Bax or Ad.Luc. Forty-eight hours after treatment, proteins were extracted from cells for western blot analysis. β-Actin was used as the internal standard.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2963730&req=5

fig1: Construction and expression of Ad.IL-24-Bax. (a) Construction of recombinant adenovirus vector that encodes the IL-24-Bax. (b) Western blot analysis of the expression of the IL-24-Bax fusion protein. Hep3B cells were treated with Ad.IL-24-Bax, Ad.IL-24, Ad.Bax or Ad.Luc. Forty-eight hours after treatment, proteins were extracted from cells for western blot analysis. β-Actin was used as the internal standard.
Mentions: As Figure 1a shows, the recombinant adenovirus IL-24-Bax was constructed as well as Ad.IL-24, Ad.Bax and Ad.Luc. When Hep3B cells were transfected with Ad.IL-24-Bax, the transfection efficiency was related to the MOI of the recombinant adenovirus. As the MOI increased, the efficiency of transfection also increased. When MOI was higher than 25, the efficiency of transfection was as high as 90%. Considering the cytotoxicity of the virus, an MOI of 10 for the virus was used in transfection experiments in this study.

Bottom Line: The mechanism of this response may include the effect of the 10HRE/VEGF385 promoter and the synergistic effect of IL-24 and Bax.Ad.IL-24-Bax also suppressed tumor growth in nude mice and induced apoptosis.Ad.IL-24-Bax may be a useful tool for gene therapy of hepatic cancer.

View Article: PubMed Central - PubMed

Affiliation: Molecular Oncology Laboratory, Eastern Hepatobiliary Surgery Hospital, Second Military Medical University, Shanghai, China.

ABSTRACT
Gene therapy promises to become an alternative choice for the treatment of hepatic cancer. In many cancers, the delivery of chimeric proteins by adenovirus vector has been reported to induce apoptosis. This study was performed to evaluate whether the recombinant adenovirus interleukin (IL)-24-Bax can induce apoptosis in hepatocellular carcinoma cells in vitro and in vivo. Several recombinant adenoviruses were constructed, and the expression of their encoded proteins was measured. The effects of the recombinant adenovirus on hepatocellular carcinoma cells and the normal hepatocyte cell line were investigated through cell viability and apoptosis assays after the cells were treated with Ad.Luc, Ad.IL-24, Ad.Bax or Ad.IL-24-Bax. The mechanism involved was also explored. A tumor-bearing mouse model was used to evaluate the effects of the adenovirus on tumor volume and cell apoptosis in vivo. Ad.IL-24-Bax selectively suppressed growth of hepatocellular carcinoma cells and induced apoptosis, but it had little influence on the normal hepatocytes. The mechanism of this response may include the effect of the 10HRE/VEGF385 promoter and the synergistic effect of IL-24 and Bax. Ad.IL-24-Bax also suppressed tumor growth in nude mice and induced apoptosis. Ad.IL-24-Bax may be a useful tool for gene therapy of hepatic cancer.

Show MeSH
Related in: MedlinePlus