[1]张淼,郭燕丽,王洛夫,等.靶向于雄激素受体的siRNA筛选及效能评价[J].陆军军医大学学报(原第三军医大学学报),2011,33(23):2459-2462.
 Zhang Miao,Guo Yanli,Wang Luofu,et al.Screening and evaluation of a newly designed siRNA targeting androgen receptor[J].J Amry Med Univ (J Third Mil Med Univ),2011,33(23):2459-2462.
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靶向于雄激素受体的siRNA筛选及效能评价(/HTML )
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陆军军医大学学报(原第三军医大学学报)[ISSN:1000-5404/CN:51-1095/R]

卷:
33卷
期数:
2011年第23期
页码:
2459-2462
栏目:
论著
出版日期:
2011-12-15

文章信息/Info

Title:
Screening and evaluation of a newly designed siRNA targeting androgen receptor
作者:
张淼郭燕丽王洛夫李彦李锐
第三军医大学西南医院:超声科,中心实验室;第三军医大学大坪医院野战外科研究所泌尿外科
Author(s):
Zhang Miao Guo Yanli Wang Luofu Li Yan Li Rui
Department of Ultrasonography, Laboratory Center of Medical Research, Southwest Hospital, Third Military Medical University, Chongqing, 400038; Department of Urology, Institute of Surgery Research, Daping Hospital,Third Military Medical University, Chongqing, 400042, China
关键词:
RNA干扰雄激素受体雄激素非依赖性前列腺癌
Keywords:
RNA interference androgen receptor androgen-independent prostate carcinoma
分类号:
R394-33; R394.3; R937.25
文献标志码:
A
摘要:
目的      设计并合成新的靶向于雄激素受体(AR)的siRNA序列,转染前列腺癌细胞筛选最佳干扰序列。      方法       设计并合成靶向于雄激素受体的siRNA,以不同浓度转染雄激素非依赖性前列腺癌细胞C4-2,确定最佳转染浓度。以最佳转染浓度转染细胞,分为siRNAⅠ组、siRNAⅡ组、siRNAⅢ组、阴性对照组、空白对照组和无关对照组。RT-PCR检测AR mRNA表达水平,Western blot检测AR蛋白表达水平。CCK-8分析绘制细胞生长曲线,观察C4-2细胞生长活性并筛选出抑制效果最佳的siRNA靶序列。      结果       以不同浓度的AR siRNA转染C4-2细胞后,细胞生长活性降低(P<0.05),并筛选出100 nmol/L的最佳转染浓度;RT-PCR和Western blot结果显示转染后AR的mRNA和蛋白表达水平显著下降(P<0.05);CCK-8法检测结果显示各组细胞增殖活性显著降低,其中siRNAⅠ的抑制效果最为明显(P<0.05)。      结论       新设计合成的AR siRNA能通过沉默AR表达高效抑制C4-2细胞的增殖。
Abstract:
Objective       To design and synthesize siRNA sequences targeting androgen receptor (AR), and to screen effective sequences by transfecting C4-2 cells.       Methods       Three pairs of siRNA sequences targeting AR were designed and synthesized according to the cDNA of AR, and were transfected into androgen-independent prostate carcinoma C4-2 cells to determine an optimal concentration. Experimental groups comprised of siRNAⅠ, siRNAⅡ, and siRNAⅢ, negative control, blank control and unrelated control. RT-PCR and Western blotting were applied to detect mRNA and protein expression of AR, respectively. Cell growth activity and the most effective siRNA sequence were evaluated based on cell growth curves by CCK-8 analysis.       Results       Cell growth activities of C4-2 cells transfected with different concentrations of AR siRNA (20 nmol/L, 50 nmol/L, 100 nmol/L and 200 nmol/L, respectively) were suppressed, and 100 nmol/L was the optimum concentration of AR siRNA (P<0.05). Compared with those of the control groups, mRNA and protein levels of AR decreased significantly, and C4-2 cell growth activity was suppressed significantly in the groups of C4-2 cells transfected with AR siRNA (P<0.05). The inhibitory effect of AR siRNAⅠ was the most significant than that of the others.       Conclusion       The newly designed AR siRNA can inhibit C4-2 cell proliferation by silencing AR expression.

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更新日期/Last Update: 2011-11-24