MYB is a novel regulator of pancreatic tumour growth and metastasis.
MYB is a novel regulator of pancreatic tumour growth and metastasis.
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DOI：
10.1038/bjc.2015.400
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发表时间：
2015-12-22
影响因子：
8.8
通讯作者：
Singh AP
中科院分区：
文献类型：
--
作者：
Srivastava SK;Bhardwaj A;Arora S;Singh S;Azim S;Tyagi N;Carter JE;Wang B;Singh AP
MYB编码一种转录因子，调节参与细胞功能的多种基因的表达。据报道，它在胰腺癌（PC）病例的子集中扩增;然而，迄今为止，其病理生物学相关性仍不清楚。通过免疫印迹或qRT-PCR分析来分析MYB和其他细胞蛋白的表达。MYB在非表达（BxPC 3）PC细胞中稳定过表达，在高表达（MiaPaCa和Panc 1）PC细胞中沉默。以24小时间隔通过自动细胞计数分析对生长的影响。在用各自的生物标志物染色后，通过流式细胞术测量具有改变的MYB表达的PC细胞的细胞周期进展和凋亡指数。在Boyden室测定中使用未包被或基质胶包被的膜检查细胞运动性/侵袭。通过非侵入性成像和通过在裸鼠胰腺中植入的内切酶标记的MYB改变的PC的终点测量来检查对致瘤性和转移潜力的影响。MYB在所有胰腺癌组织中均有异常表达，而在正常胰腺组织中未检测到。除BxPC 3外，所有测试的建立的PC细胞系也表现出MYB表达。MYB在BxPC 3细胞中的强制表达促进了它们的生长、细胞周期进展、存活和恶性行为，而其在MiaPaCa和Panc 1细胞中的沉默产生了匡威的效果。更重要的是，异位MYB表达足以赋予非致瘤性BxPC 3细胞致瘤性和转移性能力，而其沉默导致MYB过表达细胞中的显著丧失，如原位小鼠模型中所示。我们还确定了PC细胞中的几个MYB调控基因，这些基因可能潜在地介导其对肿瘤生长和转移的影响。MYB在PC细胞中异常过表达，并作为胰腺肿瘤生长和转移的关键决定因素。
MYB encodes for a transcription factor regulating the expression of a wide array of genes involved in cellular functions. It is reported to be amplified in a sub-set of pancreatic cancer (PC) cases; however, its pathobiological association has remained unclear thus far. Expression of MYB and other cellular proteins was analysed by immunoblot or qRT-PCR analyses. MYB was stably overexpressed in non-expressing (BxPC3) and silenced in highly expressing (MiaPaCa and Panc1) PC cells. Effect on growth was analysed by automated cell counting at 24-h interval. Cell-cycle progression and apoptotic indices of PC cells with altered MYB expression were measured through flow cytometry upon staining with respective biomarkers. Cell motility/invasion was examined in a Boyden's chamber assay using non-coated or Matrigel-coated membranes. Effect on tumorigenicity and metastatic potential was examined by non-invasive imaging and through end-point measurements of luciferase-tagged MYB-altered PC implanted in the pancreas of nude mice. MYB was aberrantly expressed in all malignant cases of pancreas, whereas remained undetectable in normal pancreas. All the tested established PC cell lines except BxPC3 also exhibited MYB expression. Forced expression of MYB in BxPC3 cells promoted their growth, cell-cycle progression, survival and malignant behaviour, whereas its silencing in MiaPaCa and Panc1 cells produced converse effects. More importantly, ectopic MYB expression was sufficient to confer tumorigenic and metastatic capabilities to non-tumorigenic BxPC3 cells, while its silencing resulted in significant loss of the same in MYB-overexpressing cells as demonstrated in orthotopic mouse model. We also identified several MYB-regulated genes in PC cells that might potentially mediate its effect on tumour growth and metastasis. MYB is aberrantly overexpressed in PC cells and acts as a key determinant of pancreatic tumour growth and metastasis.