Anti-tumor phosphatase and tensin homolog deleted on chromosome ten (PTEN)-VP22 gene
A PTEN-VP22, anti-tumor technology, applied in the field of genetic engineering, can solve the problems of low PTEN gene transduction efficiency, hindering the clinical application of PTEN gene therapy, etc.
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Embodiment 1
[0041] Embodiment 1—construct anti-tumor PTEN-VP22 gene by way of PCR
Embodiment 2
[0072] The PTEN-VP22 gene of the control example 1-3 obtained in embodiment 2-embodiment 1 is for the cell in vitro experiment of tumor cell
[0073] The PTEN-VP22 gene of Control Example 1-3, as well as PTEN and VP22 genes were connected into eukaryotic expression plasmid pcDNA3 to construct pcDNA3-PTEN-VP22, pcDNA3-PTEN and pcDNA3-VP22 expression vectors respectively.
[0074] Cell experiment 1: Inhibitory effect on breast cancer cell line BT549
[0075] A. Time-effect relationship: Transfect 1 μg of pcDNA3, pcDNA3-PTEN, pcDNA3-VP22, and pcDNA3-PTEN-VP22 plasmids into BT549 cells cultured in a 24-well plate, and set up a negative control group of untransfected BT549 cells. 8 replicate wells. Cells were cultured for 4 h after transfection, and 100 μl of cell suspension (1×10 3 cells), at 37°C, 5% CO 2 Cultivate under conditions; after 24h, 48h, 72h, 84h, and 96h, add 10 μl of CCK-8 solution to each well at 37°C, 5% CO 2 After incubation for 2 hours, the A450 value was mea...
Embodiment 3
[0118] The PTEN-VP22 gene of control example 1 obtained in embodiment 3-embodiment 1 is to the animal experiment of tumor cell
[0119] The PTEN-VP22 gene and PTEN of Control Example 1 were connected into the eukaryotic expression plasmid pcDNA3 to construct pcDNA3-PTEN-VP22 and pcDNA3-PTEN expression vectors respectively.
[0120] Animal Experiment 1: Inhibitory Effect on Breast Cancer Cells
[0121] Breast cancer cells BT549 were inoculated subcutaneously in nude mice to establish tumor-bearing nude mice, and the tumor-bearing nude mice were randomly divided into three groups: experimental group 1, experimental group 2 and negative control group. Experimental group 1 was injected intratumorally with 500uL, 100μg / mL pcDNA3-PTEN-VP22 gene expression vector, experimental group 2 was injected with 500uL, 100μg / mL pcDNA3-PTEN gene expression vector, and the negative control group was not injected. Observe the survival rate and average tumor volume of the three groups of mice on ...
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Abstract
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Application Information
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