PDHA1促进三阴性乳腺癌细胞的增殖、侵袭和转移

PDHA1 promotes proliferation,invasion and metastasis of triple-nega⁃tive breast cancer cells

目的:三阴性乳腺癌(triple-negative breast cancer,TNBC)的发生发展重要特征之一是细胞代谢异常。本研究旨在探究丙酮酸脱氢酶复合体中的关键酶组分丙酮酸脱氢酶E1亚基α1(pyruvate dehydrogenase E1 subunit alpha 1,PDHA1)影响TNBC增殖、转移和侵袭能力的分子机制。方法:(1)运用UALCAN数据库、KM-plotter数据库、String数据库和TCGA数据库分析PDHA1在乳腺癌组织和癌旁组织的表达水平,根据肿瘤病理分期、亚型分型和乳腺癌生物标志物的表达,比较PDHA1的表达情况,并且富集PDHA1在TNBC中的相关信号通路。(2)进行免疫组化实验检测PDHA1在人TNBC癌旁和肿瘤组织样本的表达情况。(3)构建稳定敲除PDHA1和回补PDHA1的TNBC细胞系MDA-MB-231。运用集落形成实验和细胞计数实验检测PDHA1对MDA-MB-231细胞增殖的影响。使用划痕实验和Transwell方法检测PDHA1对MDA-MB-231细胞侵袭和迁移的调节作用。通过Western blot实验,研究PDHA1缺失对MDA-MB-231细胞上皮-间充质转化通路相关蛋白的影响。结果:在数据库分析显示,乳腺癌高表达PDHA1组的生存期较短,预后较差。在临床样本中,肿瘤组织的PDHA1表达高于癌旁正常组织。敲除PDHA1会抑制MDA-MB-231细胞的增殖、转移与侵袭,抑制MDA-MB-231细胞上皮-间充质转化。结论:PDHA1在TNBC中过表达,促进TNBC细胞系MDA-MB-231的增殖、转移和侵袭。

AIM:One of the important characteristics of the occurrence and development of triple-negative breast cancer(TNBC)is dysregulated cell metabolism.The aim of this study is to investigate the mechanism of pyruvate dehydrogenase E1 subunit alpha 1(PDHA1),a key enzyme component in aerobic glycolysis,affecting the proliferation,metastasis and invasion of TNBC.METHODS:(1)The expression levels of PDHA1 in breast cancer tissues and adjacent tissues were analyzed by UALCAN database,KM-plotter database,Gene MANIA database and TCGA database.The expression of PDHA1 was compared according to tumor pathological stage,subtype classification and breast cancer biomarkers.The function of PDHA1 in TNBC was explored by gene enrichment analysis.(2)Immunohistochemistry assays were used to detect the expression of PDHA1 in human TNBC tissue and adjacent tissue samples.(3)Stable PDHA1 knockout and PDHA1 rescue TNBC MDA-MB-231 cells were constructed.The proliferation of MDA-MB-231 cells was de-tected by colony formation assay and cell counting assay.The regulatory effect of PDHA1 on the invasion and migration of MDA-MB-231 cells was detected by in vitro scratch assay and Transwell migration assay.RESULTS:Database analysis showed that the group with high PDHA1 expression in breast cancer had shorter survival and worse prognosis.In clinical specimens,the expression of PDHA1 in cancer tissues was higher than that in adjacent normal tissues.Knockout of PDHA1 inhibited the proliferation,metastasis,invasion and epithelial-mesenchymal transition of MDA-MB-231 cells.CONCLUSION:PDHA1 is overexpressed in TNBC,and it promotes cell proliferation and facilitates TNBC metastasis through the epithelial-mesenchymal transition pathway.