Apoptosis-inducing effect of vp3 gene on human breast carcinoma
Qifa Zhang
Abstract
Qifa Zhang
Abstract
Objective To construct recombinant plasmid pcDvp3 and observe the apoptosis-inducing effect of vp3 gene on human breast cancer cell line-435. Methods (1) vp3 gene was cloned into the plasmid pcDNA3.1 to form the recombinant plasmid pcDvp3. Then the nucleotides sequencing was processed. (2) 48h after transfection of pcDvp3 and pcDNA3.1 into breast cancer cell lines-435,optical microscopy, electric-microscopy, agarose electrophoresis and flow cytometry were used to verify apoptosis of tumor cells.(3)Nude mouse model of human breast cancer cells -435 was established to observe the tumor-inhibiting rate and TUNEL was adopted to identify apoptosis. Results (1) Sequence analysis justified the recombination of plasmid pcDvp3. (2) 48h after transfection into breast cancer cells-435, distinct morphological transformation and typical apoptosis bodies were observed, agarose electrophoresis of genomic DNA showed typical ladder-like pattern and flow cytometry analysis showed apoptosis peaks with the percentage of 14.42%. (3) Tumor-inhibiting rates in pcDvp3 groups were 65.52% and 68.23%, much higher than that in pcDNA3.1 group(t=4.06,P0.01), and cellular apoptosis could be seen by TUNEL assay. Conclusion vp3 gene could induce apoptosis in breast cancer cell-435 both in vitro and in vivo.
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Objective To construct recombinant plasmid pcDvp3 and observe the apoptosis-inducing effect of vp3 gene on human breast cancer cell line-435. Methods (1) vp3 gene was cloned into the plasmid pcDNA3.1 to form the recombinant plasmid pcDvp3. Then the nucleotides sequencing was processed. (2) 48h after transfection of pcDvp3 and pcDNA3.1 into breast cancer cell lines-435,optical microscopy, electric-microscopy, agarose electrophoresis and flow cytometry were used to verify apoptosis of tumor cells.(3)Nude mouse model of human breast cancer cells -435 was established to observe the tumor-inhibiting rate and TUNEL was adopted to identify apoptosis. Results (1) Sequence analysis justified the recombination of plasmid pcDvp3. (2) 48h after transfection into breast cancer cells-435, distinct morphological transformation and typical apoptosis bodies were observed, agarose electrophoresis of genomic DNA showed typical ladder-like pattern and flow cytometry analysis showed apoptosis peaks with the percentage of 14.42%. (3) Tumor-inhibiting rates in pcDvp3 groups were 65.52% and 68.23%, much higher than that in pcDNA3.1 group(t=4.06,P0.01), and cellular apoptosis could be seen by TUNEL assay. Conclusion vp3 gene could induce apoptosis in breast cancer cell-435 both in vitro and in vivo.
Key concepts: Apoptosis, Agarose gel electrophoresis, Transfection, Molecular biology, Flow cytometry, TUNEL assay, Plasmid, Recombinant DNA