Construction of a CD147 Lentiviral Expression Vector and Establishment of Its Stably Transfected A549 Cell Line
Background and objective CD147, a type of transmembrane glycoprotein embedded on the surface of tumor cells, can promote tumor invasion and metastasis. This aim of this study is to construct a CD147 lentiviral expression vector, establish its stably transfected A549 cell line, and observe the effect...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | zho |
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Chinese Anti-Cancer Association; Chinese Antituberculosis Association
2012-12-01
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| Series: | Chinese Journal of Lung Cancer |
| Subjects: | |
| Online Access: | http://dx.doi.org/10.3779/j.issn.1009-3419.2012.12.03 |
| _version_ | 1829113915026440192 |
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| author | Shaoxing YANG Chuanhao TANG Sihan WANG Santai SONG Xiaoqing LIU |
| author_facet | Shaoxing YANG Chuanhao TANG Sihan WANG Santai SONG Xiaoqing LIU |
| author_sort | Shaoxing YANG |
| collection | DOAJ |
| description | Background and objective CD147, a type of transmembrane glycoprotein embedded on the surface of tumor cells, can promote tumor invasion and metastasis. This aim of this study is to construct a CD147 lentiviral expression vector, establish its stably transfected A549 cell line, and observe the effect of CD147 on MMP-9 proliferation as well as on the invasive ability of human lung adenocarcinoma cells. Methods Full-length CD147 gene was amplified by real-time polymerase chain reaction (RT-PCR), inserted into a pEGFP vector to construct pEGFP-CD147 and pEGFP vectors, and then transfected into 293FT cells to precede the lentivirus equipment package. Subsequently, we collected the lentivirus venom to infect the A549 cells and establish a stable, overexpressed cell line named A549-CD147. The mRNA expression of MMP-9 was examined by RT-PCR. The proliferation and invasive ability of the human lung cancer cells before and after transfection were examined by the CCK-8 and Transwell methods. Results A CD147 lentiviral expression vector (pEGFP-CD147) was successfully constructed by restrictive enzyme digestion and plasmid sequencing. RT-PCR and Western blot analyses revealed increased mRNA and protein expression of CD147 gene in cells transfected with pEGFP-CD147 compared with the control groups. Therefore, the A549-CD147 cell line was successfully established through the experiment. The mRNA expression of MMP-9 also significantly increased after the upregulation of CD147 expression. Meanwhile, CCK-8 and Transwell assays indicated that the proliferation and invasive ability significantly increased in the A549-CD147 cells. Conclusion A lentiviral CD147 expression vector and its A549 cell line (A549-CD14) were successfully constructed. CD147 overexpression upregulated the protein expression of MMP-9, and strengthened the proliferation and invasive ability of human lung adenocarcinoma cells. |
| first_indexed | 2024-12-12T15:45:19Z |
| format | Article |
| id | doaj.art-9e7da3ac165f492e8b4f5ccf33efb4ec |
| institution | Directory Open Access Journal |
| issn | 1009-3419 1999-6187 |
| language | zho |
| last_indexed | 2024-12-12T15:45:19Z |
| publishDate | 2012-12-01 |
| publisher | Chinese Anti-Cancer Association; Chinese Antituberculosis Association |
| record_format | Article |
| series | Chinese Journal of Lung Cancer |
| spelling | doaj.art-9e7da3ac165f492e8b4f5ccf33efb4ec2022-12-22T00:19:46ZzhoChinese Anti-Cancer Association; Chinese Antituberculosis AssociationChinese Journal of Lung Cancer1009-34191999-61872012-12-01151269470010.3779/j.issn.1009-3419.2012.12.03Construction of a CD147 Lentiviral Expression Vector and Establishment of Its Stably Transfected A549 Cell LineShaoxing YANGChuanhao TANGSihan WANGSantai SONGXiaoqing LIUBackground and objective CD147, a type of transmembrane glycoprotein embedded on the surface of tumor cells, can promote tumor invasion and metastasis. This aim of this study is to construct a CD147 lentiviral expression vector, establish its stably transfected A549 cell line, and observe the effect of CD147 on MMP-9 proliferation as well as on the invasive ability of human lung adenocarcinoma cells. Methods Full-length CD147 gene was amplified by real-time polymerase chain reaction (RT-PCR), inserted into a pEGFP vector to construct pEGFP-CD147 and pEGFP vectors, and then transfected into 293FT cells to precede the lentivirus equipment package. Subsequently, we collected the lentivirus venom to infect the A549 cells and establish a stable, overexpressed cell line named A549-CD147. The mRNA expression of MMP-9 was examined by RT-PCR. The proliferation and invasive ability of the human lung cancer cells before and after transfection were examined by the CCK-8 and Transwell methods. Results A CD147 lentiviral expression vector (pEGFP-CD147) was successfully constructed by restrictive enzyme digestion and plasmid sequencing. RT-PCR and Western blot analyses revealed increased mRNA and protein expression of CD147 gene in cells transfected with pEGFP-CD147 compared with the control groups. Therefore, the A549-CD147 cell line was successfully established through the experiment. The mRNA expression of MMP-9 also significantly increased after the upregulation of CD147 expression. Meanwhile, CCK-8 and Transwell assays indicated that the proliferation and invasive ability significantly increased in the A549-CD147 cells. Conclusion A lentiviral CD147 expression vector and its A549 cell line (A549-CD14) were successfully constructed. CD147 overexpression upregulated the protein expression of MMP-9, and strengthened the proliferation and invasive ability of human lung adenocarcinoma cells.http://dx.doi.org/10.3779/j.issn.1009-3419.2012.12.03CD147Lentiviral expression vectorTransfectionProliferation |
| spellingShingle | Shaoxing YANG Chuanhao TANG Sihan WANG Santai SONG Xiaoqing LIU Construction of a CD147 Lentiviral Expression Vector and Establishment of Its Stably Transfected A549 Cell Line Chinese Journal of Lung Cancer CD147 Lentiviral expression vector Transfection Proliferation |
| title | Construction of a CD147 Lentiviral Expression Vector and Establishment of Its Stably Transfected A549 Cell Line |
| title_full | Construction of a CD147 Lentiviral Expression Vector and Establishment of Its Stably Transfected A549 Cell Line |
| title_fullStr | Construction of a CD147 Lentiviral Expression Vector and Establishment of Its Stably Transfected A549 Cell Line |
| title_full_unstemmed | Construction of a CD147 Lentiviral Expression Vector and Establishment of Its Stably Transfected A549 Cell Line |
| title_short | Construction of a CD147 Lentiviral Expression Vector and Establishment of Its Stably Transfected A549 Cell Line |
| title_sort | construction of a cd147 lentiviral expression vector and establishment of its stably transfected a549 cell line |
| topic | CD147 Lentiviral expression vector Transfection Proliferation |
| url | http://dx.doi.org/10.3779/j.issn.1009-3419.2012.12.03 |
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