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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psilenc-er2.1-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,G0/G1期细胞比例增加[(57.0±0.8)%vs(63.5±0.7)%,P<0.01],而S期和G2/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 investigate the effect of siRNA on the proliferation of mouse melanoma B16-F1 cells induced by ornithine decarboxylase antizyme inhibitor-1 (OAZI-1). Methods: The plasmid pSilencer2.1-U6 / OAZI-1 and the control psilenc-er2.1-U6 / scrambled plasmid were constructed and transfected into B16-F1 cells by lipofectamine The expression of OAZI-1 in B16-F1 cells was detected by blotting and quantitative PCR. The effects of psilencer2.1-U6 / OAZI-1 on the proliferation and cell cycle of B16-F1 cells were detected by MTT assay and flow cytometry. MTT assay was used to detect the effect of OAZI-1 on the anti-tumor drug docetaxel Sensitivity. RESULTS: The B16-F1 cells (B16 / OAZI-1 cells) stably transfected with psilencer2.1-U6 / OAZI-1 plasmid were successfully obtained. The expression of OAZI-1 mRNA and protein in B16 / OAZI-1 cells was negative / scrambled cells by 25.0% and 18.9%. The interference of OAZI-1 expression inhibited the proliferation of B16-F1 cells. The proportion of cells in G0 / G1 phase increased (57.0 ± 0.8% vs 63.5 ± 0.7%, P <0.01) The proportion decreased (31.5 ± 0.7)% vs (27.5 ± 0.3)%, P <0.05; (11.5 ± 0.3)% vs (9.1 ± 0.6)%, P <0.01]. Interfering OAZI-1 expression decreased the sensitivity of B16-F1 cells to paclitaxel. Conclusion: siRNA interference OAZI-1 expression can inhibit melanoma B16-F1 cell proliferation, and reduce its sensitivity to paclitaxel.