骨形态发生蛋白4信号通路能够通过诱导SNAIL2，在巴雷特食管和食管腺癌中诱导出类似上皮-间质转化的表型。
BMP4 Signaling Is Able to Induce an Epithelial-Mesenchymal Transition-Like Phenotype in Barrett's Esophagus and Esophageal Adenocarcinoma through Induction of SNAIL2.
作者信息
Kestens Christine, Siersema Peter D, Offerhaus G Johan A, van Baal Jantine W P M
机构信息
Department of Gastroenterology and Hepatology, University Medical Center Utrecht, Utrecht, the Netherlands.
Department of Gastroenterology and Hepatology, Radboud University Medical Center, Nijmegen, The Netherlands.
出版信息
PLoS One. 2016 May 18;11(5):e0155754. doi: 10.1371/journal.pone.0155754. eCollection 2016.
BACKGROUND
Bone morphogenetic protein 4 (BMP4) signaling is involved in the development of Barrett's esophagus (BE), a precursor of esophageal adenocarcinoma (EAC). In various cancers, BMP4 has been found to induce epithelial-mesenchymal transition (EMT) but its function in the development of EAC is currently unclear.
AIM
To investigate the expression of BMP4 and several members of the BMP4 pathway in EAC. Additionally, to determine the effect of BMP4 signaling in a human Barrett's esophagus (BAR-T) and adenocarcinoma (OE33) cell line.
METHODS
Expression of BMP4, its downstream target ID2 and members of the BMP4 pathway were determined by Q-RT-PCR, immunohistochemistry and Western blot analysis using biopsy samples from EAC patients. BAR-T and OE33 cells were incubated with BMP4 or the BMP4 antagonist, Noggin, and cell viability and migration assays were performed. In addition, expression of factors associated with EMT (SNAIL2, CDH1, CDH2 and Vimentin) was evaluated by Q-RT-PCR and Western blot analysis.
RESULTS
Compared to squamous epithelium (SQ), BMP4 expression was significantly upregulated in EAC and BE. In addition, the expression of ID2 was significantly upregulated in EAC and BE compared to SQ. Western blot analysis confirmed our results, showing an upregulated expression of BMP4 and ID2 in both BE and EAC. In addition, more phosphorylation of SMAD1/5/8 was observed. BMP4 incubation inhibited cell viability, but induced cell migration in both BAR-T and OE33 cells. Upon BMP4 incubation, SNAIL2 expression was significantly upregulated in BAR-T and OE33 cells while CDH1 expression was significantly downregulated. These results were confirmed by Western blot analysis.
CONCLUSION
Our results indicate active BMP4 signaling in BE and EAC and suggest that this results in an invasive phenotype by inducing an EMT-like response through upregulation of SNAIL2 and subsequent downregulation of CDH1.
背景
骨形态发生蛋白4（BMP4）信号通路参与了食管腺癌（EAC）的前体巴雷特食管（BE）的发展过程。在各种癌症中，已发现BMP4可诱导上皮-间质转化（EMT），但其在EAC发展中的作用目前尚不清楚。
目的
研究BMP4及其信号通路中几个成员在EAC中的表达情况。此外，确定BMP4信号通路在人巴雷特食管（BAR-T）和腺癌（OE33）细胞系中的作用。
方法
使用EAC患者的活检样本，通过实时定量逆转录聚合酶链反应（Q-RT-PCR）、免疫组织化学和蛋白质印迹分析来确定BMP4及其下游靶点ID2以及BMP4信号通路成员的表达情况。将BAR-T和OE33细胞与BMP4或BMP4拮抗剂Noggin孵育，然后进行细胞活力和迁移试验。此外，通过Q-RT-PCR和蛋白质印迹分析评估与EMT相关的因子（SNAIL2、CDH1、CDH2和波形蛋白）的表达。
结果
与鳞状上皮（SQ）相比，EAC和BE中BMP4的表达显著上调。此外，与SQ相比，EAC和BE中ID2的表达也显著上调。蛋白质印迹分析证实了我们的结果，显示BE和EAC中BMP4和ID2的表达均上调。此外，还观察到更多的SMAD1/5/8磷酸化。BMP4孵育抑制了细胞活力，但在BAR-T和OE33细胞中均诱导了细胞迁移。在BMP4孵育后，BAR-T和OE33细胞中SNAIL2的表达显著上调，而CDH1的表达显著下调。这些结果通过蛋白质印迹分析得到了证实。
结论
我们的结果表明BE和EAC中存在活跃的BMP4信号通路，并提示这通过上调SNAIL2并随后下调CDH1诱导类似EMT的反应，从而导致侵袭性表型。
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