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siRNA interfering ornithine decarboxylase antizyme inhibitor-1 expression decreases proliferation of melanoma B16-F1 cells
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Abstract:
Objective : To study the effect of siRNA interference of ornithine decarboxylase antizyme inhibitor-1 (OAZI-1) expression on the proliferation of mouse melanoma B16-F1 cells. Methods: siRNA expression plasmid psilencer21-U6/OAZI-1 targeting OAZI-1 gene and control psilencer2.1-U6/scrambled plasmid were constructed and transfected into B16-F1 cells by Lipofect assay. Then the OAZI-1 expression in B16-F1 cells was examined by Western blotting analysis and real-time PCR. The effects of psilencer2.1-U6/OAZI-1 on proliferation and cell cycle of B16-F1 cells were detected by MTT and FCM, respectively. The sensitivity of B16-F1 cells to antitumor drug docetaxel was measured by MTT method after interfering OAZI-1 expression. Results: B16-F1 cells stably transfected with psilencer2.1-U6/OAZI-1 plasmid (B16/OAZI-1) were successfully obtained, and the expressions of OAZI-1 mRNA and protein in B16/OAZI-1 were 25% and 18.9% of those in control B16/scrambled cells, respectively. Interference of OAZI-1 expression inhibited the proliferation of B16-F1 cells, significantly increased the cells in G0/G1 phase (\[57.0±0.8\]% vs \[63.5±0.7\]%, P<0.01) and significantly decreased cells in S and G2/M phases (\[31.5±0.7\]% vs \[27.5±0.3\]%, P<0.05; \[11.5±0.3\]% vs \[9.1±0.6\]%, P<0.01). Interference of OAZI-1 expression also decreased the sensitivity of B16-F1 cells to the antitumor drug docetaxel. Conclusion: Interfering OAZI-1 expression can suppress the proliferation of mouse melanoma B16-F1 cells and decrease their sensitivity to docetaxel.
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Project Supported:
Project supported by the National Natural Science Foundation of China (No. 30772590)
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