[1]朱恩东,李娜,肖斌,等.miR-222在胃癌中的表达分析及其靶基因FOS的鉴定[J].第三军医大学学报,2010,32(24):2477-2581.
 Zhu Endong,Li Na,Xiao Bin,et al.Identification of FOS as a novel target of miR-222 up-regulated in gastric cancer[J].J Third Mil Med Univ,2010,32(24):2477-2581.
点击复制

miR-222在胃癌中的表达分析及其靶基因FOS的鉴定(/HTML )
分享到:

《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
32卷
期数:
2010年第24期
页码:
2477-2581
栏目:
论著
出版日期:
2010-12-30

文章信息/Info

Title:
Identification of FOS as a novel target of miR-222 up-regulated in gastric cancer
作者:
朱恩东李娜肖斌黎伯胜毛旭虎邹全明
第三军医大学医学检验系临床微生物学及免疫学教研室,国家免疫生物制品工程技术研究中心,重庆市生物制药工程技术研究中心
Author(s):
Zhu Endong Li Na Xiao Bin Li Bosheng Mao Xuhu Zou Quanming
Department of Clinical Microbiology and Immunology, College of Medical Laboratory, Third Military Medical University, Chongqing, 400038, China
关键词:
miR-222FOS胃癌
Keywords:
miR-222FOSgastric cancer
分类号:
R394-33;R394.2;R735.2
文献标志码:
A
摘要:
目的  分析miR-222在胃癌中的表达水平并鉴定miR-222的靶基因。  方法  选取第三军医大学新桥医院普外科10例患者的胃癌及远癌组织,匀浆法提取总RNA,用Real-time PCR法检测miR-222表达。利用miRNA靶基因预测数据库TargetScan,MICROCOSM及PicTar对miR-222的靶基因进行预测。化学合成包含有miR-222结合位点的靶基因 3′非编码区(3′UTR)退火引物,插入荧光素酶报告载体pMIR-REPORT及绿色荧光蛋白(GFP)报告载体pMIR-GFP-REPORT,检测荧光素酶的活性变化及观察GFP表达量,并在蛋白水平用Western blot检测miR-222对靶基因的抑制。  结果  miR-222在70%(7/10)的胃癌组织样本中表达上调。生物信息学(TargetScan,MICROCOSM及PicTar 3个miRNA靶基因预测数据库)分析可得在包括人类在内的不同物种中,FOS基因3′UTR区都有miR-222的保守结合种子序列。通过荧光素酶活性检测和GFP抑制试验可得miR-222在mRNA水平可以与FOS基因结合。Western blot结果显示miR-222可以抑制c-fos蛋白表达。  结论  miR-222在胃癌样本中表达上调,证明FOS为miR-222的一个靶基因。
Abstract:
Objective  To analyze the expression profile of miR-222 in gastric cancer and identify its target so as to lay necessary basis for research on the function of miR-222 in the initiation and development of gastric cancer.   Methods  Ten paired gastric cancer and adjacent nontumor gastric mucosa tissues were obtained from pathologically-identified patients of Xinqiao Hospital (Chongqing). The total RNA was extracted by Trizol reagent with homogenizers. Real-time PCR was performed to detect the relative level of miR-222. The potential target of miR-222 was predicted by using 3 currently available prediction programs, including TargetScan, MICROCOSM, and PicTar. FOS was chosen because it was predicted by all the 3 programs. The 3′UTR sequence of FOS was obtained by chemical synthesis. This fragment was then cloned into the pMIR-REPORT and pMIR-GFP-REPORT, FOS was validated as a target of miR-222 by diversity assay of luciferase activity and GFP expression. Western blot analysis was performed to assay if Fos was down-regulated by miR-222 at the translational level.   Results  miR-222 was upregulated in 70% gastric cancer tissues (7/10). It was found miR-222 had potential conservative target seeding sites of FOS in various species including human beings by above-mentioned 3 prediction programs. It was proved that miR-222 targeted to FOS at mRNA level by luciferase assay and GFP repression experiments, and FOS was also suppressed by miR-222 at protein level.   Conclusion  miR-21 may serve as a biomarker for gastric cancer, and miR-222 may play a role in gastric cancer by targeting FOS.

参考文献/References:

朱恩东, 李娜, 肖斌, 等. miR-222 在胃癌中的表达分析及其靶基因FOS的鉴定[J].第三军医大学学报,2010,32(24):2577-2581.

相似文献/References:

[1]陈康宁,李云峰,郑彩梅,等.蛋白激酶C激活调控FOS表达参与缺血诱导神经元凋亡的研究[J].第三军医大学学报,2002,24(12):0.[doi:10.16016/j.1000-5404.2002.12.003 ]
 CHEN Kang ning,LI Yun feng,ZHENG Cai mei.[J].J Third Mil Med Univ,2002,24(24):0.[doi:10.16016/j.1000-5404.2002.12.003 ]

更新日期/Last Update: 2010-12-22