miR-148b-3p functions as a tumor suppressor in GISTs by directly targeting KIT.
miR-148b-3p functions as a tumor suppressor in GISTs by directly targeting KIT.
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miR-148b-3p 通过直接靶向 KIT 作为 GIST 中的肿瘤抑制因子
DOI：
10.1186/s12964-018-0228-z
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发表时间：
2018-04-16
期刊：
影响因子：
--
通讯作者：
Wang G
中科院分区：
文献类型：
--
作者：
Wang Y;Li J;Kuang D;Wang X;Zhu Y;Xu S;Chen Y;Cheng H;Zhao Q;Duan Y;Wang G
功能获得突变和KIT过度表达是胃肠道间质瘤的特征。MiRNA表达异常可能导致KIT过表达和肿瘤的发生。应用miRNA微阵列分析和实时定量聚合酶链式反应检测69例临床标本中的miRNA表达谱，包括50例CD117IHC+/KIT突变GIST和19例CD117IHC−/野生型GIST。GO浓缩和KEGG通路分析揭示了调控异常的miRNAs的预测靶点。通过生物信息学分析预测了与KIT表达呈负相关的异常miRNAs，并通过荧光素酶报告实验进行了验证。用细胞计数试剂盒8(CCK-8)和流式细胞仪检测细胞增殖、细胞周期停滞和细胞凋亡。伤口愈合和穿孔试验用于评估移行和侵袭。建立BALB/c裸鼠移植瘤模型，观察移植瘤的体内成瘤情况。Western印迹和qRT-PCR检测KIT及其下游效应蛋白ERK、AKT和STAT3的蛋白和mRNA水平。在与KIT表达呈负相关的6个miRNAs中，我们发现在CD117IHC+/KIT突变的GIST队列中miR-148B-3p显著下调。该miRNA随后被发现抑制GIST882细胞的增殖、迁移和侵袭。从机制上讲，miR-148B-3p通过直接与KIT mRNA的3‘-UTR结合来调节KIT的表达。在GIST882细胞中恢复miR-148B-3p的表达导致KIT及其下游效应蛋白ERK、AKT和STAT3的表达降低。然而，KIT的过表达逆转了miR-148B-3p对细胞增殖、迁移和侵袭的抑制作用。此外，我们发现miR-148B-3p表达降低与GIST患者的总体生存率(OS)和无病生存率(DFS)较差相关。MIR-148B-3p是KIT表达的重要调节因子，也是GIST潜在的预后生物标志物。
Gain-of-function mutations and overexpression of KIT are characteristic features of gastrointestinal stromal tumor (GIST). Dysregulation in miRNA expression may lead to KIT overexpression and tumorigenesis. miRNA microarray analysis and real-time PCR were used to determine the miRNA expression profiles in a cohort of 69 clinical samples including 50 CD117IHC+/KITmutation GISTs and 19 CD117IHC−/wild-type GISTs. GO enrichment and KEGG pathway analyses were performed to reveal the predicted targets of the dysregulated miRNAs. Of the dysregulated miRNAs whose expression was inversely correlated with that of KIT miRNAs were predicted by bioinformatics analysis and confirmed by luciferase reporter assay. Cell counting kit-8 (CCK-8) and flow cytometry were used to measure the cell proliferation, cycle arrest and apoptosis. Wound healing and transwell assays were used to evaluate migration and invasion. A xenograft BALB/c nude mouse model was applied to investigate the tumorigenesis in vivo. Western blot and qRT-PCR were used to investigate the protein and mRNA levels of KIT and its downstream effectors including ERK, AKT and STAT3. Of the six miRNAs whose expression was inversely correlated with that of KIT, we found that miR-148b-3p was significantly downregulated in the CD117IHC+/KITmutation GIST cohort. This miRNA was subsequently found to inhibit proliferation, migration and invasion of GIST882 cells. Mechanistically, miR-148b-3p was shown to regulate KIT expression through directly binding to the 3’-UTR of the KIT mRNA. Restoration of miR-148b-3p expression in GIST882 cells led to reduced expression of KIT and the downstream effectors proteins ERK, AKT and STAT3. However, overexpression of KIT reversed the inhibitory effect of miR-148b-3p on cell proliferation, migration and invasion. Furthermore, we found that reduced miR-148b-3p expression correlated with poor overall survival (OS) and disease-free survival (DFS) in GIST patients. miR-148b-3p functions as an important regulator of KIT expression and a potential prognostic biomarker for GISTs.
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影响因子：
64.8
作者：
Lu, J;Getz, G;Golub, TR
通讯作者：
Golub, TR
DOI：
10.1093/jnci/djr130
发表时间：
2011-06-01
期刊：
JOURNAL OF THE NATIONAL CANCER INSTITUTE
影响因子：
--
作者：
Pantaleo, Maria A.;Astolfi, Annalisa;Biasco, Guido
通讯作者：
Biasco, Guido
影响因子：
6.4
作者：
Pantaleo, Maria A.;Astolfi, Annalisa;Biasco, Guido
通讯作者：
Biasco, Guido
影响因子：
168.9
作者：
Joensuu, Heikki;Hohenberger, Peter;Corless, Christopher L.
通讯作者：
Corless, Christopher L.
影响因子：
8
作者：
Kang, HJ;Nam, SW;Kim, HG
通讯作者：
Kim, HG