WNT5A inhibits metastasis and alters splicing of Cd44 in breast cancer cells.
WNT5A inhibits metastasis and alters splicing of Cd44 in breast cancer cells.
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DOI：
10.1371/journal.pone.0058329
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Serra R
中科院分区：
文献类型：
--
作者：
Jiang W;Crossman DK;Mitchell EH;Sohn P;Crowley MR;Serra R
Wnt5a是一个非规范信号Wnt。WNT5A低表达与乳腺癌患者预后不良相关。高侵袭性乳腺癌细胞系MDA-MB-231和4T1表达极低水平的WNT5A。为了确定WNT5A的表达增强是否会影响转移行为，我们从4T1和MDA-MB-231亲本细胞系中产生了表达WNT5A的细胞。与对照组相比，表达WNT5A的细胞呈现鹅卵石状形态，体外迁移减少。细胞生长没有改变。与4t1载体对照相比，表达4T1-WNT5A的细胞通过尾静脉注射向肺转移的减少。为了确定WNT5A对转移的作用机制，我们采用微阵列和全转录组序列分析（RNA-seq）比较了4T1-WNT5A和4t1载体细胞中的基因表达。分析表明，与细胞运动相关的基因表达发生了非常显著的变化。通过半定量RT-PCR验证了WNT5A表达细胞中这些基因子集Mmp13、Nos2、Il1a、Cxcl2和Lamb3的下调。包括Cd44在内的细胞运动相关基因的转录物剪接也存在显著差异。Cd44是一种具有复杂基因组结构的粘附分子。可变外显子的使用与转移表型相关。RT-PCR证实了WNT5A表达细胞中Cd44的选择性添加。我们认为，WNT5A通过调控转录物水平和Cd44等关键基因的剪接，下调多种细胞运动通路，从而抑制肿瘤转移。
Wnt5a is a non-canonical signaling Wnt. Low expression of WNT5A is correlated with poor prognosis in breast cancer patients. The highly invasive breast cancer cell lines, MDA-MB-231 and 4T1, express very low levels of WNT5A. To determine if enhanced expression of WNT5A would affect metastatic behavior, we generated WNT5A expressing cells from the 4T1 and MDA-MB-231 parental cell lines. WNT5A expressing cells demonstrated cobblestone morphology and reduced in vitro migration relative to controls. Cell growth was not altered. Metastasis to the lung via tail vein injection was reduced in the 4T1-WNT5A expressing cells relative to 4T1-vector controls. To determine the mechanism of WNT5A action on metastasis, we performed microarray and whole-transcriptome sequence analysis (RNA-seq) to compare gene expression in 4T1-WNT5A and 4T1-vector cells. Analysis indicated highly significant alterations in expression of genes associated with cellular movement. Down-regulation of a subset of these genes, Mmp13, Nos2, Il1a, Cxcl2, and Lamb3, in WNT5A expressing cells was verified by semi-quantitative RT-PCR. Significant differences in transcript splicing were also detected in cell movement associated genes including Cd44. Cd44 is an adhesion molecule with a complex genome structure. Variable exon usage is associated with metastatic phenotype. Alternative spicing of Cd44 in WNT5A expressing cells was confirmed using RT-PCR. We conclude that WNT5A inhibits metastasis through down-regulation of multiple cell movement pathways by regulating transcript levels and splicing of key genes like Cd44.
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DOI：
10.1158/1541-7786.mcr-09-0528
发表时间：
2010-07
期刊：
Molecular cancer research : MCR
影响因子：
--
作者：
Lapuk A;Marr H;Jakkula L;Pedro H;Bhattacharya S;Purdom E;Hu Z;Simpson K;Pachter L;Durinck S;Wang N;Parvin B;Fontenay G;Speed T;Garbe J;Stampfer M;Bayandorian H;Dorton S;Clark TA;Schweitzer A;Wyrobek A;Feiler H;Spellman P;Conboy J;Gray JW
通讯作者：
Gray JW
影响因子：
15.9
作者：
Glynn, Sharon A.;Boersma, Brenda J.;Ambs, Stefan
通讯作者：
Ambs, Stefan
影响因子：
64.5
作者：
Acharyya S;Oskarsson T;Vanharanta S;Malladi S;Kim J;Morris PG;Manova-Todorova K;Leversha M;Hogg N;Seshan VE;Norton L;Brogi E;Massagué J
通讯作者：
Massagué J
DOI：
10.1023/b:jomg.0000037157.94207.33
发表时间：
2004-04-01
影响因子：
2.5
作者：
Brennan, KR;Brown, AMC
通讯作者：
Brown, AMC
影响因子：
15.9
作者：
Brown, Rhonda L.;Reinke, Lauren M.;Cheng, Chonghui
通讯作者：
Cheng, Chonghui