CREB1通过LAYN介导的TLN1/β1整合素轴的转录调控促进胆管癌转移：CREB1通过调节LAYN/TLN1/β1整合素轴促进胆管癌转移。

CREB1 promotes cholangiocarcinoma metastasis through transcriptional regulation of the LAYN-mediated TLN1/β1 integrin axis: CREB1 promotes cholangiocarcinoma metastasis through regulating LAYN/TLN1/β1 integrin axis.

作者信息

Liang Shuai, Zhou Shuhua, Tang Yangshuo, Xiao Moyan, Ye Ke

机构信息

Department of Pancreatic Surgery, Xiangya Hospital, Central South University, Changsha, 410008, China.

Comprehensive Surgery, Xiangya Boai Rehabilitation Hospital, Changsha, 410100, China.

出版信息

Heliyon. 2024 Aug 22;10(17):e36595. doi: 10.1016/j.heliyon.2024.e36595. eCollection 2024 Sep 15.

DOI:10.1016/j.heliyon.2024.e36595

PMID:39286102

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11402943/

Abstract

BACKGROUND

Layilin (LAYN) plays an important role in tumor progression, invasion, and metastasis; however, its role in cholangiocarcinoma (CHOL) has not been elucidated.

METHODS

We utilized the GEPIA, STRING, and hTFtarget databases for bioinformatics analysis. Overexpression or knockdown cell lines were constructed by transfecting the cells with different plasmids. Western blot (WB) was performed to detect LAYN, TLN1, and CREB1 expression. Cell proliferation, migration, and invasiveness were assessed using CCK-8 and Transwell assays. Immunofluorescence and WB were used to detect epithelial-mesenchymal transition (EMT) markers. The CHOL metastasis model was established by injecting RBE cells into the tail veins of nude mice. Metastatic lesions were identified using hematoxylin and eosin staining. Co-immunoprecipitation and Chromatin immunoprecipitation were used to validate the interactions.

RESULTS

LAYN was highly expressed in the CHOL cells. Knockdown of LAYN significantly inhibited proliferation, migration, invasion, and EMT in both QBC-939 and RBE human CHOL cells, while overexpression of LAYN had the opposite effect. Furthermore, in a CHOL metastasis model using nude mice, knocking down LAYN expression markedly suppressed CHOL liver and lung metastases. LAYN interacts with TLN1, and CREB1 binds to the LAYN promoter, with all three showing a positive correlation. Additionally, bioinformatics analysis revealed high expression of both TLN1 and CREB1 in CHOL. Knockdown of TLN1 or CREB1 in QBC-939 and RBE cells inhibited cell proliferation, migration, invasion, and EMT, reversing the effects of LAYN overexpression. Moreover, knockdown of TLN1 or CREB1 also suppressed the expression of ITGB1 and the phosphorylation levels of c-Jun, p38 MAPK, and ERK, further reversing the effects of LAYN overexpression.

CONCLUSION

Our results suggest that CREB1 promotes CHOL metastasis through transcriptional regulation of the LAYN-mediated TLN1/β1 integrin axis.

摘要

背景

层粘连蛋白受体（LAYN）在肿瘤进展、侵袭和转移中起重要作用；然而，其在胆管癌（CHOL）中的作用尚未阐明。

方法

我们利用GEPIA、STRING和hTFtarget数据库进行生物信息学分析。通过用不同质粒转染细胞构建过表达或敲低细胞系。进行蛋白质免疫印迹（WB）检测LAYN、纽带蛋白1（TLN1）和环磷腺苷效应元件结合蛋白1（CREB1）的表达。使用细胞计数试剂盒-8（CCK-8）和Transwell实验评估细胞增殖、迁移和侵袭能力。采用免疫荧光和WB检测上皮-间质转化（EMT）标志物。通过将人胆管癌细胞系RBE细胞注射到裸鼠尾静脉建立胆管癌转移模型。使用苏木精-伊红染色鉴定转移病灶。采用免疫共沉淀和染色质免疫沉淀验证相互作用。

结果

LAYN在胆管癌细胞中高表达。敲低LAYN可显著抑制人胆管癌细胞系QBC-939和RBE的增殖、迁移、侵袭和EMT，而过表达LAYN则产生相反的效果。此外，在使用裸鼠的胆管癌转移模型中，敲低LAYN表达可显著抑制胆管癌肝转移和肺转移。LAYN与TLN1相互作用，CREB1与LAYN启动子结合，三者均呈正相关。此外，生物信息学分析显示TLN1和CREB1在胆管癌中均高表达。敲低QBC-939和RBE细胞中的TLN1或CREB1可抑制细胞增殖、迁移、侵袭和EMT，逆转LAYN过表达的作用。此外，敲低TLN1或CREB1还可抑制整合素β1（ITGB1）的表达以及c-Jun、p38丝裂原活化蛋白激酶（MAPK）和细胞外信号调节激酶（ERK）的磷酸化水平，进一步逆转LAYN过表达的作用。