miRNA-183 suppresses apoptosis and promotes proliferation in esophageal cancer by targeting PDCD4.
miRNA-183 suppresses apoptosis and promotes proliferation in esophageal cancer by targeting PDCD4.
复制标题
miRNA-183 通过靶向 PDCD4 抑制食管癌细胞凋亡并促进增殖
DOI：
10.14348/molcells.2014.0147
复制
发表时间：
2014-12-31
影响因子：
3.8
通讯作者：
Wang Y
中科院分区：
文献类型：
--
作者：
Yang M;Liu R;Li X;Liao J;Pu Y;Pan E;Yin L;Wang Y
在我们先前的研究中，miR-96-182-183簇中的miRNA-183在食管鳞癌(ESCC)中显著过表达。在本研究中，我们通过对食道癌细胞系EC9706的功能得失分析，探讨miR-183在食管鳞癌中的致癌作用。应用全基因组mRNA芯片确定miR-183直接或间接调控的基因。采用3‘端非编码区荧光素酶报告基因分析、RT-PCR和Western印迹方法对miR-183目的基因进行验证。细胞培养结果显示，miR-183抑制细胞凋亡(p<；0.05)，促进细胞增殖(p<；0.05)，促进G1/S转换(p<；0.05)。此外，当miR-183被miR-183抑制剂抑制时，miR-183对细胞凋亡的抑制作用被挽救(p<；0.05)。Western印迹分析显示，当miR-183过表达时，被基因芯片和生物信息学预测为miR-183靶基因的程序性细胞死亡4(PDCD4)的表达降低。3‘端非编码区荧光素酶报告实验证实miR-183通过与PDCD4的3’端非编码区序列结合而直接调控PDCD4。Pearson相关分析进一步证实了miR-183与PDCD4在两种细胞系和ESCC患者中的显著负相关。我们的数据提示miR-183可能通过调节PDCD4的表达而在ESCC中发挥致癌作用。
In our previous study, miRNA-183, a miRNA in the miR-96-182-183 cluster, was significantly over-expressed in esophageal squamous cell carcinoma (ESCC). In the present study, we explored the oncogenic roles of miR-183 in ESCC by gain and loss of function analysis in an esophageal cancer cell line (EC9706). Genome-wide mRNA microarray was applied to determine the genes that were regulated directly or indirectly by miR-183. 3′UTR luciferase reporter assay, RT-PCR, and Western blot were conducted to verify the target gene of miR-183. Cell culture results showed that miR-183 inhibited apoptosis (p < 0.05), enhanced cell proliferation (p < 0.05), and accelerated G1/S transition (p < 0.05). Moreover, the inhibitory effect of miR-183 on apoptosis was rescued when miR-183 was suppressed via miR-183 inhibitor (p < 0.05). Western blot analysis showed that the expression of programmed cell death 4 (PDCD4), which was predicted as the target gene of miR-183 by microarray profiling and bioinformatics predictions, decreased when miR-183 was over-expressed. The 3′UTR luciferase reporter assay confirmed that miR-183 directly regulated PDCD4 by binding to sequences in the 3′UTR of PDCD4. Pearson correlation analysis further confirmed the significant negative correlation between miR-183 and PDCD4 in both cell lines and in ESCC patients. Our data suggest that miR-183 might play an oncogenic role in ESCC by regulating PDCD4 expression.