文章摘要
熊兵红,马 利,程 勇,张才全.miR-21在大肠癌中的表达及其靶基因探讨[J].南京医科大学学报,2014,(3):281~286
miR-21在大肠癌中的表达及其靶基因探讨
The expression of miR-21 in colorectal cancer and its target gene study
投稿时间:2013-10-18  
DOI:10.7655/NYDXBNS20140302
中文关键词: 大肠癌  miR-21  肿瘤抑制基因  PTEN  Akt  MMP-9
英文关键词: colorectal cancer  miR-21  tumor suppressor gene  PTEN  Akt  MMP-9
基金项目:重庆市自然科学基金资助项目(csts2012jjA0038)
作者单位
熊兵红 重庆医科大学附属第一医院普外科,普外科学重点实验室,重庆 400016 
马 利 重庆市花溪医院内科,重庆 400054 
程 勇 重庆医科大学附属第一医院普外科,普外科学重点实验室,重庆 400016 
张才全 重庆医科大学附属第一医院普外科,普外科学重点实验室,重庆 400016 
摘要点击次数: 981
全文下载次数: 413
中文摘要:
      目的:探讨微小RNA(microRNA,miRNA)-21在大肠癌(colorectal cancer,CRC)组织和细胞中的表达,抑制miRNA-21(miR-21)表达对PI3K/PTEN/AKT/mTOR信号通路中各蛋白的影响及miR-21可能调控的靶基因?方法:运用实时荧光定量PCR和免疫组化分别测定CRC组织中的miR-21和PTEN水平?通过反义寡核苷酸技术抑制大肠癌细胞株HCT116中miR-21的表达并加以验证?通过蛋白质免疫印迹法检测HCT116细胞中PTEN?PI3K?Akt?MMP-9?mTOR表达的改变?利用生物信息学分析预测miRNA-21可能的靶基因,荧光素酶双报告基因实验检测PTEN活性?结果:miR-21在CRC组织中增高(P < 0.05),PTEN在CRC组织中表达减低(P < 0.05)?而且 miR-21与PTEN的表达之间呈负相关(r = -0.396,P < 0.05)?miR-21在HCT116中表达最高,在SW480中表达最低?miRNA-21在细胞株HCT116中的表达得到有效抑制,并且PTEN蛋白的表达明显上调(P < 0.05),PI3K?Akt?MMP-9?mTOR表达下调(P < 0.05)?筛选并确认了PTEN为miR-21的调控靶基因之一?结论:PTEN可作为miR-21的靶基因参与调控CRC过程?miR-21在大肠癌中表达增高,通过抑制PTEN表达促进肿瘤细胞侵袭转移?
英文摘要:
      Objective:To investigate the expression of microRNA-21 (miR-21) in colorectal cancer (CRC) tissues and cells,and investigate the inhibition of miR-21 on protein expression of PI3K/ PTEN/AKT/ mTOR signaling pathway,and look for the target genes which may be regulated by miR-21. Methods:The expressions of miR-21 and PTEN from tissues of CRC were detected by real-time fluorescence quantitative PCR (FQ –PCR) and immunohistochemistry (IHC). The anti-sense oligonucleotide (ASO) technology was performed to inhibit and verify the expression of miR-21 in HCT116 cells. The changes in PTEN,PI3K,Akt,MMP-9 and mTOR protein expressions were detected by Western blotting. The potential target gene prediction was performed by Bioinformatics. Dual luciferase report gene assay was performed to identify the activity of PTEN. Results:The expression level of miR-21 was increased in CRC tissue (P < 0.05),and the expression of PTEN was decreased in CRC tissue (P < 0.05). However,the expression level of miR-21 was negatively associated with PTEN expression (r = -0.396,P < 0.05). The level of miR-21 was the highest in HCT116 cell and lowest in SW480 cell. The expression of miRNA-21 in HCT116 cell lines was significantly inhibited,the expression of PTEN protein was significantly increased and the expressions of PI3K,Akt,MMP-9 and mTOR were down-regulated. PTEN was identified as one of target genes of miR-21 in CRC cell. Conclusion:PTEN,as one of target genes of miR-21 in CRC cell,can regulate the process of CRC. miR-21 expression is increased in CRC,and promotes cancer cell invasion and metastasis by inhibiting PTEN.
查看全文   查看/发表评论  下载PDF阅读器
关闭