Construction of recombinant lentiviral vectors containing Rheb gene and its mutant Rheb'D60K gene and their expression in human liver cancer cells / 南方医科大学学报
Journal of Southern Medical University
; (12): 341-344, 2012.
Article
in Zh
| WPRIM
| ID: wpr-267604
Responsible library:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To construct recombinant lentiviral vectors carrying Rheb gene and its mutant Rheb'D60K gene, and examine their expression in human liver cancer cells.</p><p><b>METHODS</b>Rheb gene was amplified by PCR to construct the recombinant plasmid LV31-Rheb-WT and LV31-Rheb-D60K. HEK-293 FT cells were contransfected with the recombinant lentiviral vector together with a lentiviral package plasmid to produce the lentiviral particles. The expression of PS6 protein was detected in the lentivirus-infected MCF-7 cells. The apoptosis of SK-HEP-1 cells transfected with LV31-Rheb-WT or LV31-Rheb-D60K was observed.</p><p><b>RESULTS</b>The recombinant LV31-Rheb-WT and LV31-Rheb-D60K vectors were confirmed by PCR and DNA sequencing. Western blotting showed that PS6 protein expression was increased in LV31-Rheb-WT-transfected cells while decreased in LV31-Rheb-D60K-transfected cells. LV31-Rheb-D60K-transfected SK-HEP-1 cells showed more obvious apoptosis after starvation than LV31-Rheb-WT-transfected cells.</p><p><b>CONCLUSION</b>Lentiviral vectors carrying Rheb gene and its mutant has been successfully constructed, which can be useful in further investigation of the role of Rheb gene in cancer cells.</p>
Full text:
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Index:
WPRIM
Main subject:
Pathology
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Neuropeptides
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Recombinant Proteins
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Transfection
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Apoptosis
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Carcinoma, Hepatocellular
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Lentivirus
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Monomeric GTP-Binding Proteins
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Mutant Proteins
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HEK293 Cells
Limits:
Humans
Language:
Zh
Journal:
Journal of Southern Medical University
Year:
2012
Type:
Article