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A novel Bifidobacterium infantis-mediated TK/GCV suicide gene therapy system exhibits antitumor activity in a rat model of bladder cancer.

Tang W, He Y, Zhou S, Ma Y, Liu G - J. Exp. Clin. Cancer Res. (2009)

Bottom Line: Successful clinical management remains a challenge.The engineered BI-TK was injected into tumor-bearing rats via tail vein, followed by intraperitoneal injection of ganciclovir (GCV).Immunohistochemical staining further demonstrated that the BI-TK/GCV treatment group had the highest level of caspase3 protein expression than that of the empty plasmid group and normal saline group (p < 0.05).

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Urology, First Affiliated Hospital, Chongqing Medical University, Chongqing, China. tangwei2060@yahoo.com.cn

ABSTRACT
Bladder cancer is the ninth most common malignancy in the world. Successful clinical management remains a challenge. In order To search for novel targeted and efficacious treatment, we sought to investigate anti-tumor activity of BI-TK suicide gene therapy system in a rat model of bladder tumors. We first constructed and tested an anaerobic Bifidobacterium infantis-mediated thymidine kinase (BI-TK) suicide gene therapy system. To test the in vivo efficacy of this system, we established a rat model of bladder tumors, which was induced by N-methyl-nitrosourea perfusion. Bifidobacterium infantis containing the HSV-TK (i.e., BI-TK) were constructed by transformation of recombinant plasmid pGEX - TK. The engineered BI-TK was injected into tumor-bearing rats via tail vein, followed by intraperitoneal injection of ganciclovir (GCV). Using the rat model of bladder tumors, we found that bladder tumor burdens were significantly lower in the rats treated with BI-TK/GCV group than that treated with normal saline control group (p <0.05). While various degrees of apoptosis of the tumor cells were detected in all groups using in situ TUNEL assay, apoptosis was mostly notable in the BI-TK/GCV treatment group. Immunohistochemical staining further demonstrated that the BI-TK/GCV treatment group had the highest level of caspase3 protein expression than that of the empty plasmid group and normal saline group (p < 0.05). Thus, our results demonstrate that the Bifidobacterium infantis-mediated TK/GCV suicide gene therapy system can effectively inhibit rat bladder tumor growth, possibly through increasing caspase 3 expression and inducing apoptosis.

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Construction and verification of Bifidobacterium infantis-mediated TK tumor-targeting suicide gene therapy system. Plasmid DNA was purified from anaerobic culture, digested with restriction enzymes, and resolved on 1% agarose gel. The expected 6.0 kb fragment of pGEX-TK is indicated by arrows.
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Figure 1: Construction and verification of Bifidobacterium infantis-mediated TK tumor-targeting suicide gene therapy system. Plasmid DNA was purified from anaerobic culture, digested with restriction enzymes, and resolved on 1% agarose gel. The expected 6.0 kb fragment of pGEX-TK is indicated by arrows.

Mentions: The pGEX - TK recombinant vector was transformed into Bifidobacterium infantis by electroporation, After being cultured for 72 hours, Bifidobacterium infantis formed scattered colonies on the LB-plates containing MRS and ampicillin LB-plates. In contrast, transformatoion wild-type Bifidobacterium infantis only had no colonies on the MRS benzyl penicillin LB plates. Single colonies were picked up and grown under anaerobic condition. DNA was purified and verified by restriction enzymatic digestion and PCR amplification (Figure 1).


A novel Bifidobacterium infantis-mediated TK/GCV suicide gene therapy system exhibits antitumor activity in a rat model of bladder cancer.

Tang W, He Y, Zhou S, Ma Y, Liu G - J. Exp. Clin. Cancer Res. (2009)

Construction and verification of Bifidobacterium infantis-mediated TK tumor-targeting suicide gene therapy system. Plasmid DNA was purified from anaerobic culture, digested with restriction enzymes, and resolved on 1% agarose gel. The expected 6.0 kb fragment of pGEX-TK is indicated by arrows.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2803447&req=5

Figure 1: Construction and verification of Bifidobacterium infantis-mediated TK tumor-targeting suicide gene therapy system. Plasmid DNA was purified from anaerobic culture, digested with restriction enzymes, and resolved on 1% agarose gel. The expected 6.0 kb fragment of pGEX-TK is indicated by arrows.
Mentions: The pGEX - TK recombinant vector was transformed into Bifidobacterium infantis by electroporation, After being cultured for 72 hours, Bifidobacterium infantis formed scattered colonies on the LB-plates containing MRS and ampicillin LB-plates. In contrast, transformatoion wild-type Bifidobacterium infantis only had no colonies on the MRS benzyl penicillin LB plates. Single colonies were picked up and grown under anaerobic condition. DNA was purified and verified by restriction enzymatic digestion and PCR amplification (Figure 1).

Bottom Line: Successful clinical management remains a challenge.The engineered BI-TK was injected into tumor-bearing rats via tail vein, followed by intraperitoneal injection of ganciclovir (GCV).Immunohistochemical staining further demonstrated that the BI-TK/GCV treatment group had the highest level of caspase3 protein expression than that of the empty plasmid group and normal saline group (p < 0.05).

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Urology, First Affiliated Hospital, Chongqing Medical University, Chongqing, China. tangwei2060@yahoo.com.cn

ABSTRACT
Bladder cancer is the ninth most common malignancy in the world. Successful clinical management remains a challenge. In order To search for novel targeted and efficacious treatment, we sought to investigate anti-tumor activity of BI-TK suicide gene therapy system in a rat model of bladder tumors. We first constructed and tested an anaerobic Bifidobacterium infantis-mediated thymidine kinase (BI-TK) suicide gene therapy system. To test the in vivo efficacy of this system, we established a rat model of bladder tumors, which was induced by N-methyl-nitrosourea perfusion. Bifidobacterium infantis containing the HSV-TK (i.e., BI-TK) were constructed by transformation of recombinant plasmid pGEX - TK. The engineered BI-TK was injected into tumor-bearing rats via tail vein, followed by intraperitoneal injection of ganciclovir (GCV). Using the rat model of bladder tumors, we found that bladder tumor burdens were significantly lower in the rats treated with BI-TK/GCV group than that treated with normal saline control group (p <0.05). While various degrees of apoptosis of the tumor cells were detected in all groups using in situ TUNEL assay, apoptosis was mostly notable in the BI-TK/GCV treatment group. Immunohistochemical staining further demonstrated that the BI-TK/GCV treatment group had the highest level of caspase3 protein expression than that of the empty plasmid group and normal saline group (p < 0.05). Thus, our results demonstrate that the Bifidobacterium infantis-mediated TK/GCV suicide gene therapy system can effectively inhibit rat bladder tumor growth, possibly through increasing caspase 3 expression and inducing apoptosis.

Show MeSH
Related in: MedlinePlus