Analysis of MiR-195 and MiR-497 Expression, Regulation and Role in Breast Cancer
Analysis of MiR-195 and MiR-497 Expression, Regulation and Role in Breast Cancer
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DOI：
10.1158/1078-0432.ccr-10-1800
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发表时间：
2011-04-01
影响因子：
11.5
通讯作者：
Lai, Lihui
中科院分区：
文献类型：
--
作者：
Li, Dan;Zhao, Yulan;Lai, Lihui
目的：为探讨miR-195和miR-497在乳腺癌中的表达、调控、潜在作用和靶点，实验设计：采用北方印迹和实时荧光定量PCR方法初步检测miR-195和miR-497在乳腺癌组织和细胞系中的表达模式。采用亚硫酸氢盐限制性酶切分析和亚硫酸氢盐测序相结合的方法研究miR-195和miR-497基因的DNA甲基化状态。建立稳定表达miR-195和miR-497的乳腺癌细胞，以研究其作用和靶点。结果：miR-195和miR-497在乳腺癌组织中表达显著下调，在乳腺癌组织中表达显著下调，miR-195和miR-497在乳腺癌组织中表达显著下调。发现miR-195/497基因上游CpG岛的甲基化状态负责两种miRNA的下调。miR-195或miR-497的强制表达抑制乳腺癌细胞增殖和侵袭。Raf-1和Ccnd 1被鉴定为miR-195和miR-497的新的直接靶点。结论：miR-195和miR-497在乳腺癌中的表达水平与乳腺癌的恶性程度呈负相关。临床癌症研究; 17（7）; 1722-30。(C)2011年AACR。
Purpose: To investigate expression, regulation, potential role and targets of miR-195 and miR-497 in breast cancer.Experimental Design: The expression patterns of miR-195 and miR-497 were initially examined in breast cancer tissues and cell lines by Northern blotting and quantitative real-time PCR. Combined bisulfite restriction analysis and bisulfite sequencing were carried out to study the DNA methylation status of miR195 and miR-497 genes. Breast cancer cells stably expressing miR-195 and miR-497 were established to study their role and targets. Finally, normal, fibroadenoma and breast cancer tissues were employed to analyze the correlation between miR-195/497 levels and malignant stages of breast tumor tissues.Results: MiR-195 and miR-497 were significantly downregulated in breast cancer. The methylation state of CpG islands upstream of the miR-195/497 gene was found to be responsible for the downregulation of both miRNAs. Forced expression of miR-195 or miR-497 suppressed breast cancer cell proliferation and invasion. Raf-1 and Ccnd1 were identified as novel direct targets of miR-195 and miR-497. miR-195/497 expression levels in clinical specimens were found to be correlated inversely with malignancy of breast cancer.Conclusions: Our data imply that both miR-195 and miR-497 play important inhibitory roles in breast cancer malignancy and may be the potential therapeutic and diagnostic targets. Clin Cancer Res; 17(7); 1722-30. (C) 2011 AACR.