A TrkB-STAT3-miR-204-5p regulatory circuitry controls proliferation and invasion of endometrial carcinoma cells.
A TrkB-STAT3-miR-204-5p regulatory circuitry controls proliferation and invasion of endometrial carcinoma cells.
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DOI：
10.1186/1476-4598-12-155
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发表时间：
2013-12-09
期刊：
影响因子：
37.3
通讯作者：
Wan XP
中科院分区：
文献类型：
--
作者：
Bao W;Wang HH;Tian FJ;He XY;Qiu MT;Wang JY;Zhang HJ;Wang LH;Wan XP
我们以前确定TrkB作为参与促进子宫内膜癌（EC）转移的癌基因。在这里，我们试图描绘TrkB表达的变化对EC细胞中microRNA（miRNA）的整体概况的影响，并进一步研究某些miRNA和TrkB在EC患者的临床病理特征中的表达之间的相关性。采用定量逆转录-PCR（qRT-PCR）技术，我们发现TrkB mRNA的表达在正常子宫内膜和激光捕获显微切割的EC细胞中的转录水平没有显著差异，而免疫组化结果显示TrkB蛋白在EC组织中的表达明显高于正常子宫内膜。microRNA阵列显示，异位过表达和TrkB表达的敲低导致EC细胞中miRNA表达的整体变化。qRT-PCR结果显示升高的TrkB抑制EC细胞中miR-204- 5 p的表达。此外，免疫印迹分析显示，TrkB在IshikawaTrkB细胞中的过表达显著增加了JAK 2和STAT 3的磷酸化，然而，在HEC-1BshTrkB细胞中，TrkB的敲低使JAK 2和STAT 3的磷酸化中止。此外，ChIP分析显示磷酸化STAT 3可以直接结合到miR-204- 5 p上游TRPM 3启动子区域附近的STAT 3结合位点。有趣的是，使用生物信息学分析和荧光素酶测定，我们确定TrkB是miR-204- 5 p的新靶点。MTT法、克隆形成法和Transwell法显示miR-204- 5 p在功能上显著抑制EC细胞的克隆形成生长、迁移和侵袭。此外，miR-204- 5 p还抑制携带人EC细胞的肿瘤异种移植物的生长。重要的是，我们发现较低的miR-204- 5 p表达与晚期FIGO分期、淋巴结转移相关，并且可能降低EC患者的生存机会。这项研究揭示了一个涉及TrkB/miR-204- 5 p的新的调控环，该环对EC的肿瘤发生至关重要，并提出miR-204- 5 p表达的重建可以作为这种疾病的潜在新治疗靶点进行探索。
We previously identified TrkB as an oncogene involved in promoting metastasis in endometrial carcinoma (EC). Here, we sought to delineate the effect of changes in TrkB expression on the global profile of microRNAs (miRNAs) in EC cells and further investigated the correlation between the expression of certain miRNA and TrkB in the clinicopathologic characteristics of EC patients. Using quantitative reverse transcription-PCR (qRT-PCR), we found that expression of TrkB mRNA has no significant difference in transcript levels between normal endometrium and EC cells captured by laser capture microdissection, while immunohistochemistry results demonstrated a markedly higher expression of TrkB protein in EC tissues. The microRNA array showed that ectopic overexpression and knockdown of TrkB expression caused global changes in miRNA expression in EC cells. qRT-PCR results showed that elevated TrkB repressed miR-204-5p expression in EC cells. Furthermore, immunoblotting assays revealed that TrkB overexpression in IshikawaTrkB cells noticeably increased JAK2 and STAT3 phosphorylation, which, however, was aborted by TrkB knockdown in HEC-1BshTrkB cells. Moreover, ChIP assays showed that phospho-STAT3 could directly bind to STAT3-binding sites near the TRPM3 promoter region upstream of miR-204-5p. Interestingly, using bioinformatics analysis and luciferase assays, we identified TrkB was a novel target of miR-204-5p. Functionally, the MTT assays, clonogenic and Transwell assays showed that miR-204-5p significantly suppressed the clonogenic growth, migration and invasion of EC cells. Furthermore, miR-204-5p also inhibited the growth of tumor xenografts bearing human EC cells. Importantly, we found lower miR-204-5p expression was associated with advanced FIGO stages, lymph node metastasis and probably a lower chance for survival in EC patients. This study uncovers a new regulatory loop involving TrkB/miR-204-5p that is critical to the tumorigenesis of EC and proposes that reestablishment of miR-204-5p expression could be explored as a potential new therapeutic target for this disease.
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影响因子：
4.7
作者：
Boren, Todd;Xiong, Yin;Lancaster, Johnathan M.
通讯作者：
Lancaster, Johnathan M.
影响因子：
64.8
作者：
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影响因子：
37.3
作者：
Chen Z;Sangwan V;Banerjee S;Mackenzie T;Dudeja V;Li X;Wang H;Vickers SM;Saluja AK
通讯作者：
Saluja AK
DOI：
10.1073/pnas.0914785107
发表时间：
2010-08-31
影响因子：
11.1
作者：
Conte, Ivan;Carrella, Sabrina;Banfi, Sandro
通讯作者：
Banfi, Sandro
影响因子：
3.7
作者：
Imam JS;Plyler JR;Bansal H;Prajapati S;Bansal S;Rebeles J;Chen HI;Chang YF;Panneerdoss S;Zoghi B;Buddavarapu KC;Broaddus R;Hornsby P;Tomlinson G;Dome J;Vadlamudi RK;Pertsemlidis A;Chen Y;Rao MK
通讯作者：
Rao MK