Construction and Antiapoptosis Activities of Recombinant Adenoviral Expression Vector Carrying EBV Latent Membrane Protein 2A
To evaluate the possible effects of LMP2A (EBV latent membrane protein 2A) on human gastric cancer cell line SGC-7901, LMP2A coding gene was subcloned into shuttle plasmid pAdTrackCMV to form transfer plasmid pAdTrackCMV-2A, which was linearized with PmeI and cotransformed into E.coli BJ5183 with ad...
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| Format: | Article |
| Language: | English |
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Wiley
2011-01-01
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| Series: | Gastroenterology Research and Practice |
| Online Access: | http://dx.doi.org/10.1155/2011/182832 |
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| author | Xishuang Liu Yu Gao Bing Luo Youan Zhao |
| author_facet | Xishuang Liu Yu Gao Bing Luo Youan Zhao |
| author_sort | Xishuang Liu |
| collection | DOAJ |
| description | To evaluate the possible effects of LMP2A (EBV latent membrane protein 2A) on human gastric cancer cell line SGC-7901, LMP2A coding gene was subcloned into shuttle plasmid pAdTrackCMV to form transfer plasmid pAdTrackCMV-2A, which was linearized with PmeI and cotransformed into E.coli BJ5183 with adenovirus genomic plasmid of pAdeasy-1. The identified recombinant adenovirus plasmid DNA was digested with PacI and transfected into 293 cells to package recombinant adenovirus particles named vAd-2A. Then the expression and antiapoptosis activities of LMP2A on SGC-7901 infected with vAd-2A were analyzed. The vAd-2A was successfully constructed and identified by PCR, restriction digestion, and sequencing. LMP2A expression in SGC was identified by strong green fluorescence expression with fluorescence microscopic photograph and Southern blotting. The growth of LMP2A expressing SGC cells was apparently improved. Both cyclin E expression and S phase ratio in LMP2A expressing SGC cells were upregulated by cell cycle analysis and confocal microscopic analysis respectively. The replication-deficient recombinant adenovirus vector can express LMP2A antigen in SGC cells and inhibit their apoptosis. The results indicate that LMP2A might play an important role in pathogenesis of EBV-associated gastric cancer (EBVaGC). This study establishes a foundation for further study on EBVaGC and its gene therapy. |
| format | Article |
| id | doaj-art-bbba6034dd314d35bf5cc63d9e193784 |
| institution | Kabale University |
| issn | 1687-6121 1687-630X |
| language | English |
| publishDate | 2011-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Gastroenterology Research and Practice |
| spelling | doaj-art-bbba6034dd314d35bf5cc63d9e1937842025-02-03T05:46:19ZengWileyGastroenterology Research and Practice1687-61211687-630X2011-01-01201110.1155/2011/182832182832Construction and Antiapoptosis Activities of Recombinant Adenoviral Expression Vector Carrying EBV Latent Membrane Protein 2AXishuang Liu0Yu Gao1Bing Luo2Youan Zhao3Department of Gastroenterology, Qilu Hospital, Shandong University, Ji'nan, Shandong 250012, ChinaDepartment of Medical Microbiology, Medical College, Qingdao University, Qingdao, Shandong 266021, ChinaDepartment of Medical Microbiology, Medical College, Qingdao University, Qingdao, Shandong 266021, ChinaDepartment of Gastroenterology, Qilu Hospital, Shandong University, Ji'nan, Shandong 250012, ChinaTo evaluate the possible effects of LMP2A (EBV latent membrane protein 2A) on human gastric cancer cell line SGC-7901, LMP2A coding gene was subcloned into shuttle plasmid pAdTrackCMV to form transfer plasmid pAdTrackCMV-2A, which was linearized with PmeI and cotransformed into E.coli BJ5183 with adenovirus genomic plasmid of pAdeasy-1. The identified recombinant adenovirus plasmid DNA was digested with PacI and transfected into 293 cells to package recombinant adenovirus particles named vAd-2A. Then the expression and antiapoptosis activities of LMP2A on SGC-7901 infected with vAd-2A were analyzed. The vAd-2A was successfully constructed and identified by PCR, restriction digestion, and sequencing. LMP2A expression in SGC was identified by strong green fluorescence expression with fluorescence microscopic photograph and Southern blotting. The growth of LMP2A expressing SGC cells was apparently improved. Both cyclin E expression and S phase ratio in LMP2A expressing SGC cells were upregulated by cell cycle analysis and confocal microscopic analysis respectively. The replication-deficient recombinant adenovirus vector can express LMP2A antigen in SGC cells and inhibit their apoptosis. The results indicate that LMP2A might play an important role in pathogenesis of EBV-associated gastric cancer (EBVaGC). This study establishes a foundation for further study on EBVaGC and its gene therapy.http://dx.doi.org/10.1155/2011/182832 |
| spellingShingle | Xishuang Liu Yu Gao Bing Luo Youan Zhao Construction and Antiapoptosis Activities of Recombinant Adenoviral Expression Vector Carrying EBV Latent Membrane Protein 2A Gastroenterology Research and Practice |
| title | Construction and Antiapoptosis Activities of
Recombinant Adenoviral Expression Vector Carrying
EBV Latent Membrane Protein 2A |
| title_full | Construction and Antiapoptosis Activities of
Recombinant Adenoviral Expression Vector Carrying
EBV Latent Membrane Protein 2A |
| title_fullStr | Construction and Antiapoptosis Activities of
Recombinant Adenoviral Expression Vector Carrying
EBV Latent Membrane Protein 2A |
| title_full_unstemmed | Construction and Antiapoptosis Activities of
Recombinant Adenoviral Expression Vector Carrying
EBV Latent Membrane Protein 2A |
| title_short | Construction and Antiapoptosis Activities of
Recombinant Adenoviral Expression Vector Carrying
EBV Latent Membrane Protein 2A |
| title_sort | construction and antiapoptosis activities of recombinant adenoviral expression vector carrying ebv latent membrane protein 2a |
| url | http://dx.doi.org/10.1155/2011/182832 |
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