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1.
Journal of Southern Medical University ; (12): 832-837, 2015.
Article in Chinese | WPRIM | ID: wpr-355274

ABSTRACT

<p><b>OBJECTIVE</b>To construct a RNA interference lentiviral vector for human glutathione peroxidase 2 (GPX2) gene and observe the effect of GPX2 knockdown on cell apoptosis.</p><p><b>METHODS</b>The sequence of the small interfering RNA (siRNA) for GPX2 interference was inserted into the pSicoR vector. HepG2 cells were infected by the packaged si-GPX2 lentivirus and the expression of GPX2 in the infected cells was detected by both RT-PCR and Western blotting. Changes of cell apoptosis following the infection were analyzed by flow cytometry.</p><p><b>RESULTS</b>The lentiviral particles pSicoR-GPX2 were successfully packaged. The expression of GPX2 in the infected cells was obviously down-regulated at both RNA and protein levels. GPX2 knockdown caused increased apoptosis rate, increased Bax expression and lowered Bcl-2 expression in HepG2 cells.</p><p><b>CONCLUSION</b>We have successfully constructed the lentiviral vector for RNA interference of human GPX2 gene.</p>


Subject(s)
Humans , Apoptosis , Down-Regulation , Genetic Vectors , Glutathione Peroxidase , Genetics , Hep G2 Cells , Lentivirus , RNA Interference , RNA, Small Interfering
2.
Journal of Southern Medical University ; (12): 614-617, 2012.
Article in Chinese | WPRIM | ID: wpr-267539

ABSTRACT

<p><b>OBJECTIVE</b>To construct a lentiviral vector for RNA interference (RNAi) of human glycerol kinase (GK) gene to stably down-regulate GK expression in human hepatocytes.</p><p><b>METHODS</b>The sequence of siRNA for GK interference were cloned into the pSicoR vector. Following packaging in 293T cells, the lentivirus was titrated using fluorescence activated cell sorting. Human hepatocyte L02 cells was infected with the lentivirus and the expression of GK was analyzed using Western blotting.</p><p><b>RESULTS</b>The lentiviral particle pSicoR-GK was successfully packaged with a virus titer reaching 3×10(7) pfu/ml. The expression level of GK protein was down-regulated to 20% of the control level in L02 cells infected with the lentivirus.</p><p><b>CONCLUSION</b>The lentiviral vector for RNAi of human GK gene has been successfully constructed, which can significantly down-regulate GK expression in human hepatocytes.</p>


Subject(s)
Humans , Cell Line , Gene Expression Regulation , Genetic Vectors , Glycerol Kinase , Genetics , Hepatocytes , Lentivirus , Genetics , Plasmids , RNA Interference , RNA, Small Interfering , Genetics , Transfection
3.
Acta Anatomica Sinica ; (6)2002.
Article in Chinese | WPRIM | ID: wpr-681850

ABSTRACT

Objective To study the expression of ABC gene family of MCF 7/ADM cells as compared to parental cell line MCF 7/S, and its reversion by realgar of the expressed genes. Methods Expression of ABC family genes was examined by RT PCR, and cell growth by MTT test. Results The expression of ABC gene family was positive in MCF 7/ADM cells compared to MCF 7/S cells; Realgar can down regulate the expression ABC gene family in MCF 7/ADM cells; the IC 50 of ADM to MCF 7/ADM cells was decreased.Conclusion ABC gene family was responsible for the induced resistance of MCF 7/ADM cells to ADM; Realgar is able to reverse partly the multidrug resistance of the human breast cancer cell line MCF 7/ADM. [

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