Effects of DARPP-32 on the drug sensitivity of SH-SY5Y cells |
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Authors: | ZHANG Hong-wei LIU Li-hua LI Wei |
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Affiliation: | 1.Department of Neurology, 2Laboratory of Molecular Diagnosis, First Hospital, Wenli University, Shaoxing 312000, China;3Department of Neurology, Huaxi Hospital, Sichuan University, Chengdu, 610041, China. E-mail:zhw0606@sina.com |
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Abstract: | AIM: To investigate the effects of DARPP-32 on the drug sensitivity of human neuroblastoma cell line SH-SY5Y. METHODS: The plasmid containing cDNA of DARPP-32 gene and the small interfering RNA eukaryotic expression vector specific to human DARPP-32 gene were constructed and transfected into SH-SY5Y cells. Stable transfectants were identified by RT-PCR and Western blotting analysis. MTT assay and flow cytometry assay were used to investigate the effects of DARPP-32 on the drug sensitivity and the drug accumulation in cell models. The expressions of Bcl-2,Bax, P-gp and MRP were analyzed by RT-PCR and Western blotting. RESULTS: The stable clones with increased or decreased DARPP-32 expression were successfully established. Up-regulation of DARPP-32 significantly enhanced the drug sensitivity and the drug accumulation of SH-SY5Y cells due to down-regulation of P-gp and Bcl-2 expressions. Down-regulation of DARPP-32 significantly reduced the drug sensitivity of SH-SY5Y cells and decreased drug accumulation in SH-SY5Y cells. CONCLUSION: DARPP-32 might mediate the sensitivity of SH-SY5Y cells to conventional chemotherapeutic agents. |
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Keywords: | Neuroblastoma Genes DARPP-32 RNA interference SH-SY5Y cells |
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