BMP4 通过调控 EMT 参与非综合征型唇腭裂（伴或不伴腭裂）的发生。

BMP4 Regulates EMT to be Involved in non-Syndromic Cleft lip With or Without Palate.

机构信息

Key Lab of Environment and Health, School of Public Health, Xuzhou Medical University, Jiangsu, China.

出版信息

Cleft Palate Craniofac J. 2023 Nov;60(11):1462-1473. doi: 10.1177/10556656221105762. Epub 2022 Jun 14.

DOI:10.1177/10556656221105762

PMID:35702016

Abstract

OBJECTIVE

In the previous study, we identified bone morphogenetic protein 4 (BMP4) responsible for non-syndromic cleft lip with or without cleft palate (NSCL/P). We aimed to elucidate the effects and mechanisms of BMP4 on epithelial-mesenchymal transition (EMT) through Smad1 signaling pathway to be involved in NSCL/P.

METHODS

The human oral epidermoid carcinoma cells (KBs) were transfected with plasmids or small interfering RNA (siRNA) to build the models. The migration of the cells was evaluated by transwell assay. Western blotting and quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR) were used to detect the expressions of BMP4, E-cadherin, N-cadherin, EMT-related transcription factors snal1 and snal2, matrix metalloproteinase 2 (MMP2), MMP9, Smad1, and phosphorylated Smad1.

RESULTS

In the overexpression group, the migration number of cells was increased significantly. The protein expression of E-cadherin was decreased significantly, while the protein expression level of the N-cadherin was increased significantly. The protein and mRNA expressions of MMP2, MMP9, snal1, and snal2 were significantly higher. The expression level of Smad1 was not significantly changed, while the phosphorylation of Smad1 was significantly increased. In the BMP4-siRNA group, the migrating number cells was significantly decreased. The protein expression of E-cadherin was increased significantly, while the expression of N-cadherin was significantly decreased. The protein and mRNA expressions of MMP2, MMP9, snal1, and snal2 were significantly lower than that of the control group. The expressions of Smad1 and phosphorylation of Smad1 were not significantly changed.

CONCLUSIONS

BMP4 enhances cell migration and promotes cell EMT through Smad1 signaling pathway. Abnormal BMP4 mediates migration and EMT through other relevant signaling pathways resulting in NSCL/P. The study provides new insight into the mechanisms of NSCL/P associated with BMP4.n.

摘要

目的

在之前的研究中，我们发现骨形态发生蛋白 4（BMP4）负责非综合征性唇裂伴或不伴腭裂（NSCL/P）。我们旨在通过 Smad1 信号通路阐明 BMP4 对上皮-间充质转化（EMT）的影响和机制，从而参与 NSCL/P。

方法

用人口腔表皮样癌细胞（KBs）转染质粒或小干扰 RNA（siRNA）构建模型。通过 Transwell 测定评估细胞迁移。Western blot 和实时定量逆转录聚合酶链反应（qRT-PCR）用于检测 BMP4、E-钙粘蛋白、N-钙粘蛋白、EMT 相关转录因子 snal1 和 snal2、基质金属蛋白酶 2（MMP2）、MMP9、Smad1 和磷酸化 Smad1 的表达。

结果

在过表达组中，细胞迁移数量显著增加。E-钙粘蛋白的蛋白表达显著降低，而 N-钙粘蛋白的蛋白表达水平显著升高。MMP2、MMP9、snal1 和 snal2 的蛋白和 mRNA 表达显著升高。Smad1 的表达水平没有显著变化，而 Smad1 的磷酸化显著增加。在 BMP4-siRNA 组中，细胞迁移数量显著减少。E-钙粘蛋白的蛋白表达显著增加，而 N-钙粘蛋白的表达显著降低。MMP2、MMP9、snal1 和 snal2 的蛋白和 mRNA 表达显著低于对照组。Smad1 的表达和磷酸化 Smad1 没有显著变化。