目的： 研究siRNA干扰鸟氨酸脱羧酶抗酶抑制因子-1（ornithine decarboxylase antizyme inhibitor-1， OAZI-1）的表达对小鼠黑素瘤B16-F1细胞增殖的影响。 方法： 构建OAZI-1特异性siRNA表达质粒psilencer2.1-U6/OAZI-1及对照psilencer2.1-U6/scrambled质粒，脂质体转染法将质粒转染入B16-F1细胞，Western blotting和定量PCR检测B16-F1细胞中OAZI-1的表达。MTT法和流式细胞术检测psilencer2.1-U6/OAZI-1对B16-F1细胞增殖和细胞周期的影响；MTT法检测干扰OAZI-1表达后B16-F1细胞对抗肿瘤药物紫杉醇(docetaxel)的敏感性。 结果： 成功获得稳定转染psilencer2.1-U6/OAZI-1质粒的B16-F1细胞（B16/OAZI-1细胞），B16/OAZI-1细胞中OAZI-1 mRNA和蛋白表达分别为阴性对照B16/scrambled细胞的250%和18.9%。干扰OAZI-1的表达抑制B16-F1细胞的增殖，G0/G1期细胞比例增加\[（57.0±0.8)% vs (63.5±0.7)%，P<0.01\]，而S期和G2/M期细胞比例减少\[（31.5±0.7)% vs (27.5±0.3)%，P<0.05；(11.5±0.3)% vs (9.1±0.6)%，P<0.01\]。干扰OAZI-1的表达能降低B16-F1细胞对紫杉醇的敏感性。 结论： siRNA干扰OAZI-1的表达能抑制黑素瘤B16-F1细胞的增殖，并降低其对紫杉醇的敏感性。

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 psilencer21-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 G0/G1 phase (\[57.0±0.8\]% vs \[63.5±0.7\]%, P<0.01) and significantly decreased cells in S and G2/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.

国家自然科学基金资助项目（No. 30772590）