siRNA干扰鸟氨酸脱羧酶抗酶抑制因子-1的表达抑制黑素瘤细胞增殖  

siRNA interfering ornithine decarboxylase antizyme inhibitor-1 expression decreases proliferation of melanoma B16-F1 cells

作　　者：何玲[1] 韩钰[1] 刘梦瑶[1] 蔡富强[1] 王艳林[1] 

机构地区：[1]三峡大学分子生物学研究所,湖北宜昌443002

出　　处：《中国肿瘤生物治疗杂志》2011年第4期414-418,共5页Chinese Journal of Cancer Biotherapy

基　　金：国家自然科学基金资助项目(No.30772590)~~

摘　　要：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 study the effect of siRNA interference of ornithine decarboxylase antizyme inhibitor-1 （ OA- ZI-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 BI6-F1 cells was examined by Western blotting analysis and real-time PCR. The effects of psilencer2. I-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 plas- mid （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.

关 键 词：鸟氨酸脱羧酶抗酶抑制因子-1 RNA干扰技术 黑素瘤细胞 

分 类 号：R739.5[医药卫生—肿瘤] R730.54[医药卫生—临床医学]