BRWD3对乳腺癌淋巴结转移的影响

Effect of BRWD3 on lymph node metastasis in breast cancer

目的探讨溴结构域和WD重复蛋白3(BRWD3)对乳腺癌淋巴结转移的影响及其作用机制。方法采用体外细胞侵袭实验探究BRWD3对乳腺癌细胞系侵袭能力的调控作用。采用裸鼠淋巴结转移和肺转移模型探究BRWD3对裸鼠乳腺癌淋巴结转移和肺转移的调控作用。通过cBioPortal数据库检索BRWD3共表达基因,进行生物功能及通路的聚类分析,构建BRWD3分子调控网络,并用Western blotting进行部分验证。采用公共数据库数据和KMPLOT平台分析BRWD3在乳腺癌中的表达情况及其与乳腺癌预后的关系。结果体外实验结果显示,敲除BRWD3促进了乳腺癌细胞系的侵袭(P<0.01)和迁移,但未促进乳腺癌细胞增殖。裸鼠淋巴结转移模型显示,敲除BRWD3能够明显促进乳腺癌淋巴结转移;肺转移模型显示,BRWD3未影响乳腺癌的肺转移能力。GO功能分析显示,BRWD3共表达基因主要富集于基因表达调控、DNA损伤修复、染色体的组织与修饰、蛋白质泛素化等生物学功能。KEGG富集分析显示,BRWD3共表达基因主要参与蛋白质泛素化、氧化磷酸化、MAPK等信号通路。Western blotting检测证实,敲除BRWD3促进了ERK1/2的磷酸化。BRWD3在发生淋巴结转移乳腺癌患者中的表达量明显低于未发生淋巴结转移的患者。KMPLOT分析结果显示,BRWD3低表达患者的预后较差,其生存期明显短于BRWD3高表达患者。结论 BRWD3能够调控乳腺癌的侵袭和体内淋巴结转移,BRWD3低表达患者预后较差。BRWD3可能亦具有调控蛋白质泛素化、氧化磷酸化、MAPK通路等功能。该发现为乳腺癌淋巴结转移相关分子标志物的研究和应用提供了新的理论基础。

Objective To explore the effect of the bromodomain and WD repeat domain containing 3 (BRWD3) on lymph node metastasis in breast cancer and its mechanism.Methods In vitro cell invasion experiments were used to explore the effect of BRWD3 on the invasion phenotype of breast cancer cell lines.Mouse lymph node metastasis model and lung metastasis model were used to investigate the role of BRWD3 in regulating breast cancer lymph node metastasis and lung metastasis in BALB/c nude mice.The BRWD3 co-expressed genes were searched through cBioPortal databases to analyze the biological functions and pathways of BRWD3,constructed a BRWD3 molecular regulatory network,which is examined with Western blotting analysis partly.The public breast cancer dataset and the KMPLOT analysis platform was used to analyze the expression of BRWD3 in breast cancer and the relationship between the expression of BRWD3 and breast cancer prognosis.Results In vitro experiments showed that knockdown of BRWD3 significantly enhanced the invasion and migration of breast cancer cells (P<0.01),but did not promote their proliferation.The lymph node metastasis model demonstrated that knockdown of BRWD3 dramatically enhanced lymph node metastasis of breast cancer.Interestingly,the lung metastasis model showed that BRWD3 did not affect the mouse lung metastasis ability.Further functional clustering GO analysis of the co-expressed genes of BRWD3 suggested that they are mainly involved in gene expression regulation,DNA damage repair,chromosome organization and modification,ubiquitination,etc.Meanwhile,KEGG enrichment analysis showed that they were involved in signaling pathways such as ubiquitination,oxidative phosphorylation,MAPK,etc.Besides,via the Western blotting experiment,it was found that knockdown of BRWD3 increased the phosphorylation of ERK1/2.Moreover,BRWD3 expression in breast cancer with lymph node metastasis is significantly lower than in patients without lymph node metastasis.Further,the survival analysis in KMPLOT found that the prognosis of patients with low expression of BRWD3 was poor,which was significantly lower than that of patients with high expression of BRWD3.Conclusions BRWD3 can regulate breast cancer invasion in vitro and lymph node metastasis in vivo.Afterward,the prognosis of patients with low expression of BRWD3 is poor.Meanwhile,ubiquination,oxidative phosphorylation,MAPK pathway,etc.were the possible regulation pathways of BRWD3,which provide a new theoretical basis for the research and application of molecular markers related to breast cancer lymph node metastasis.