2005Zhonghua putong waike zazhiRequires access

Apoptosis-inducing effect of vp3 gene on human breast carcinoma

Qifa Zhang

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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) 48h 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) 48h 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) 48h 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) 48h 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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Available 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) 48h 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) 48h 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

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