Yu Ruoxi, Li Zhi, Zhang Chuang, Song Huicong, Deng Mingming, Sun Liping, Xu Ling, Che Xiaofang, Hu Xuejun, Qu Xiujuan, Liu Yunpeng, Zhang Ye
Department of Medical Oncology, the First Affiliated Hospital of China Medical University, Shenyang, China.
Key Laboratory of Anticancer Drugs and Biotherapy, Shenyang, China.
PeerJ. 2019 May 6;7:e6885. doi: 10.7717/peerj.6885. eCollection 2019.
The limb-bud and heart development (LBH) gene is a highly conserved, tissue-specific transcription cofactor in vertebrates that regulates multiple key genes in embryonic development. The role of LBH in various cancer types is still controversial, and its specific role and molecular mechanism in the oncogenesis of gastric cancer (GC) remains largely unexplored. In the present study, the prognostic significance and clinicopathological characteristics of LBH in GC was determined. The LBH mRNA expression was first investigated in four independent public datasets (TCGA-STAD, GSE15459, GSE29272, and GSE62254) and then validated with our samples at the protein level. LBH was overexpressed at both the mRNA and protein levels in cancer compared with normal tissues. High LBH expression was correlated with advanced T, N, and M stages. Kaplan-Meier analysis and log-rank test indicated that higher LBH expression was statistically correlated with shorter overall survival (OS) in the public datasets and our study samples. Univariate and multivariate Cox regression analysis showed that LBH was an independent prognostic biomarker for survival in TCGA-STAD, GSE15459, GSE62254 cohorts, and our GC patients. experiments showed that knockdown of LBH can significantly inhibit the proliferation and invasion of HGC-27 cells, while overexpression of LBH can significantly enhance the proliferation and invasion of BGC-823 cells. Gene Set Enrichment Analysis (GSEA), Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomics (KEGG) indicated that high LBH expression is associated with the PI3K-Akt pathway, focal adhesion, and extracellular matrix (ECM)-receptor interaction. Western blot analysis showed that knockdown of LBH significantly inhibited the expression of integrin α5, integrin β1, p-FAK, and p-Akt. Therefore, results from the present study indicate that LBH is a potential independent prognostic biomarker and promotes proliferation and invasion of GC cells by activating the integrin/FAK/Akt pathway.
肢体芽与心脏发育（LBH）基因是脊椎动物中一种高度保守的组织特异性转录辅因子，可调节胚胎发育中的多个关键基因。LBH在各种癌症类型中的作用仍存在争议，其在胃癌（GC）发生中的具体作用和分子机制在很大程度上仍未得到探索。在本研究中，确定了LBH在GC中的预后意义和临床病理特征。首先在四个独立的公共数据集（TCGA-STAD、GSE15459、GSE29272和GSE62254）中研究LBH mRNA表达，然后在蛋白质水平上用我们的样本进行验证。与正常组织相比，癌症组织中LBH在mRNA和蛋白质水平均过表达。高LBH表达与晚期T、N和M分期相关。Kaplan-Meier分析和对数秩检验表明，在公共数据集和我们的研究样本中，较高的LBH表达与较短的总生存期（OS）在统计学上相关。单因素和多因素Cox回归分析表明，LBH是TCGA-STAD、GSE15459、GSE62254队列以及我们的GC患者生存的独立预后生物标志物。实验表明，敲低LBH可显著抑制HGC-27细胞的增殖和侵袭，而LBH的过表达可显著增强BGC-823细胞的增殖和侵袭。基因集富集分析（GSEA）、基因本体论（GO）和京都基因与基因组百科全书（KEGG）表明，高LBH表达与PI3K-Akt途径、粘着斑和细胞外基质（ECM）-受体相互作用相关。蛋白质印迹分析表明，敲低LBH可显著抑制整合素α5、整合素β1、p-FAK和p-Akt的表达。因此，本研究结果表明，LBH是一种潜在的独立预后生物标志物，并通过激活整合素/FAK/Akt途径促进GC细胞的增殖和侵袭。
