Silencing of miR-148a in cancer-associated fibroblasts results in WNT10B-mediated stimulation of tumor cell motility.
Silencing of miR-148a in cancer-associated fibroblasts results in WNT10B-mediated stimulation of tumor cell motility.
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DOI：
10.1038/onc.2012.351
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发表时间：
2013-07-04
期刊：
影响因子：
8
通讯作者：
Niederhuber, J.
中科院分区：
文献类型：
--
作者：
Aprelikova, O.;Palla, J.;Hibler, B.;Yu, X.;Greer, Y. E.;Yi, M.;Stephens, R.;Maxwell, G. L.;Jazaeri, A.;Risinger, J. I.;Rubin, J. S.;Niederhuber, J.
肿瘤微环境在癌症进展中具有重要作用。在这里，我们发现miR-148 a在16个癌症相关成纤维细胞（CAF）样本中的15个（94%）中下调，与从子宫内膜癌患者中建立的匹配的正常组织成纤维细胞（NF）相比。对正常组织和子宫内膜癌间质细胞的激光捕获显微切割证实了这一观察结果。用5-氮杂-脱氧胞苷处理细胞刺激了大多数CAF中miR-148 a的表达，暗示DNA甲基化在miR-148 a表达的调节中。对成纤维细胞中miR-148 a功能的研究表明，来自过表达miR-148 a的CAF的条件培养基（CM）显著损害了5种子宫内膜癌细胞系的迁移，而不影响它们在共培养实验中的生长速率。在预测的miR-148 a靶基因中，有两个WNT家族成员，WNT 1和WNT 10 B。CAF中WNT/β-连环蛋白途径的激活通过基因表达的微阵列分析和SuperTOPFIash荧光素酶报告基因的活性增加来证实。我们发现CAFs中WNT 10 B蛋白水平升高，当通过慢病毒感染重新引入miR-148 a时，其水平降低。当与miR-148 a共表达时，克隆在荧光素酶cDNA下游的WNT 10 B的3′-UTR抑制荧光素酶活性，表明WNT 10 B是miR-148 a的直接靶点。与miR-148 a的作用相反，WNT 10 B刺激子宫内膜癌细胞系的迁移。我们的研究结果已经确定了肿瘤微环境中的分子机制，这是癌症治疗的新靶点。
The tumor microenvironment has an important role in cancer progression. Here we show that miR-148a is downregulated in 15 out of 16 samples (94%) of cancer-associated fibroblasts (CAFs) compared with matched normal tissue fibroblasts (NFs) established from patients with endometrial cancer. Laser-capture microdissection of stromal cells from normal tissue and endometrial cancer confirmed this observation. Treatment of cells with 5-aza-deoxycytidine stimulated the expression of miR-148a in the majority of CAFs implicating DNA methylation in the regulation of miR-148a expression. Investigation of miR-148a function in fibroblasts demonstrated that conditioned media (CM) from CAFs overexpressing miR-148a significantly impaired the migration of five endometrial cancer cell lines without affecting their growth rates in co-culture experiments. Among predicted miR-148a target genes are two WNT family members, WNT1 and WNT10B. Activation of the WNT/β-catenin pathway in CAFs was confirmed by microarray analysis of gene expression and increased activity of the SuperTOPFIash luciferase reporter. We found elevated levels of WNT10B protein in CAFs and its level decreased when miR-148a was re-introduced by lentiviral infection. The 3′-UTR of WNT10B, cloned downstream of luciferase cDNA, suppressed luciferase activity when co-expressed with miR-148a indicating that WNT10B is a direct target of miR-148a. In contrast to the effect of miR-148a, WNT10B stimulated migration of endometrial cancer cell lines. Our findings have defined a molecular mechanism in the tumor microenvironment that is a novel target for cancer therapy.
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影响因子：
13.6
作者：
Di Leva G;Croce CM
通讯作者：
Croce CM
影响因子：
8
作者：
Lane, TF;Philip, L
通讯作者：
Philip, L
DOI：
10.1073/pnas.0401064101
发表时间：
2004-04-06
影响因子：
11.1
作者：
Kuperwasser, C;Chavarria, T;Weinberg, RA
通讯作者：
Weinberg, RA
影响因子：
10.5
作者：
Olson, Peter;Lu, Jun;Hanahan, Douglas
通讯作者：
Hanahan, Douglas
影响因子：
4.6
作者：
Nagendra, Deepak C.;Burke, James, III;Risinger, John I.
通讯作者：
Risinger, John I.