[1]吴开杰,张栋,曾津,等.RNAi沉默Smad4基因表达对人前列腺癌细胞增殖的影响[J].第三军医大学学报,2009,31(13):1250-1253.
 WU Kai-jie,ZHANG Dong,ZENG Jin,et al.Effects of Smad4 gene silencing by small interfering RNA on the proliferation of prostate cancer cells in vitro[J].J Third Mil Med Univ,2009,31(13):1250-1253.
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
31卷
期数:
2009年第13期
页码:
1250-1253
栏目:
专题报道
出版日期:
2009-07-15

文章信息/Info

Title:
Effects of Smad4 gene silencing by small interfering RNA on the proliferation of prostate cancer cells in vitro
作者:
吴开杰张栋曾津薛艳薛玉泉王蓉王新阳贺大林
西安交通大学医学院第一附属医院:泌尿外科,肿瘤中心,影像中心
Author(s):
WU Kai-jie ZHANG Dong ZENG Jin XUE Yan XUE Yu-quan WANG Rong WANG Xin-yang HE Da-lin
Department of Urology, Center of Oncology,Center of Radiology,  First Affiliated Hospital, Medical School of Xi′an Jiaotong University, Xi′an 710061, Shaanxi Province, China
关键词:
前列腺癌肿瘤进展RNA干扰Smad4
Keywords:
prostate cancer tumor progression RNA interference Smad4
分类号:
R394.2; R737.25;R730.231
文献标志码:
A
摘要:
目的   观察RNA干扰技术沉默Smad4基因表达对人前列腺癌细胞增殖能力的影响,探讨Smad4在前列腺癌恶性进展中的作用。   方法   用半定量RT-PCR和Western blot方法检测LNCaP、PC-3、DU145以及ARCaP亚细胞系IF11、IA8等多种不同前列腺癌细胞Smad4 mRNA及蛋白表达的情况,筛选出Smad4基因阳性表达的细胞;针对Smad4基因不同部位设计并化学合成3对不同的靶向小干扰RNA(small interfering RNA, siRNA),脂质体介导瞬时转染阳性表达Smad4的细胞,转染后48 h分别用RT-PCR和Western blot检测Smad4 mRNA及蛋白表达水平的变化,用MTT法检测沉默Smad4表达对细胞增殖的影响。   结果   检测的5种前列腺癌细胞中仅LNCaP、PC-3、DU145 3种细胞表达Smad4 mRNA及蛋白;选择PC-3细胞做siRNA转染,与阴性对照序列组(siRNA-NC)相比,转染48 h后,转染SiRNA1、SiRNA2、SiRNA3的3组PC-3细胞的Smad4 mRNA表达水平均显著下降,但仅有siRNA3组Smad4蛋白表达水平显著降低。转染SiRNA3沉默Smad4基因表达后PC-3细胞的体外增殖能力显著增强(P<0.05)。   结论   不同前列腺癌细胞Smad4基因的表达存在差异,靶向siRNA下调Smad4的表达可增强前列腺癌细胞的增殖能力。
Abstract:
Objective   To explore the effects of Smad4 gene silencing by small interfering RNA (siRNA) on the proliferation of prostate cancer cells in vitro.     Methods   Expression of Smad4 gene in several prostate cancer cell lines (LNCaP, PC-3, DU145, IF11, and IA8) was detected by semi-quantitative reverse transcription-PCR (RT-PCR) and Western blotting for the screening of the positive cells. Three siRNA targeting three different domains of Smad4 gene were designed, synthesized, and transfected into Smad4-positive cells by LipofectamineTM2000. After transfection for 48 h, expression of Smad4 gene in Smad4-positive cells was detected by RT-PCR and Western blotting again, and the proliferative potency was also analyzed by MTT assay at the same time.     Results   There was a remarkable difference in the expressions of Smad4 mRNA and protein between these cells. Smad4 was highly expressed in LNCaP, PC-3, and DU145 cells, but not in IF11 and IA8 cells. Compared with the negative control (siRNA-NC) groups, transfection of three targeting siRNA in PC-3 cells knocked down the expression of Smad4 mRNA significantly, but only siRNA3 silenced the expression of Smad4 protein. Smad4 gene silencing enhanced the proliferative capacity of PC-3 cells after transfection.     Conclusion   There is difference in the expression of Smad4 gene in prostate cancer cell lines. The chemically synthesized specific siRNA targeting Smad4 could effectively reduce the expression of Smad4 gene, and enhance the proliferative capacity of PC-3 cells in vitro.

参考文献/References:

吴开杰, 张栋, 曾津, 等.RNAi沉默Smad4基因表达对人前列腺癌细胞增殖的影响[J]. 第三军医大学学报,2009,31(13):1250-1253.

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更新日期/Last Update: 2009-06-16