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三阴性乳腺癌关键基因的生物信息学分析及癌基因PLK1的验证

Bioinformatics analysis of key genes in triple negative breast cancer and validation of oncogene PLK1.

作者信息

Ren Yi, Deng Rong, Zhang Qian, Li Jing, Han Baosan, Ye Ping

机构信息

Breast Department, Xuzhou Cancer Hospital/Xuzhou Hospital Affiliated to Jiangsu University, Xuzhou, China.

Department of General Surgery, Jiangsu Cancer Hospital, Nanjing, China.

出版信息

Ann Transl Med. 2020 Dec;8(24):1637. doi: 10.21037/atm-20-6873.

Abstract

BACKGROUND

Breast cancer is the most common malignancy in women. Triple-negative breast cancer (TNBC) refers to a special subtype that is deficient in the expression of estrogen (ER), progesterone (PR), and human epidermal growth factor receptor 2 (HER-2). In this study, a variety of bioinformatics analysis tools were used to screen Hub genes related to the occurrence and development of triple negative breast cancer, and their biological functions were analyzed.

METHODS

Gene Expression Omnibus (GEO) breast cancer microarray data GSE62931 was selected as the research object. The differentially expressed genes (DEGs) were screened and the protein-protein interaction (PPI) network of DEGs was constructed using bioinformatics tools. The Hub genes were also screened. The Gene Ontology (GO) knowledgebase and the Kyoto Encyclopedia of Genes and Genomes (KEGG) were used for biological enrichment analysis. The Gene Expression Profiling Interactive Analysis (GEPIA) online tool was used to verify the expression of the screened genes and patient survival. The effects of polo-like kinase 1 (PLK1) on the proliferation, invasion, migration, and dryness of breast cancer cells were verified using cell counting kit 8 (CCK-8), transwell migration assays, scratch tests, and clone formation tests. An animal model of subcutaneous xenotransplantation of breast cancer was established to evaluate the effect of PLK1 on the proliferation of breast cancer.

RESULTS

A total of 824 DEGs were screened by GSE62931 microarray data; 405 of which were up-regulated and 419 of which were down-regulated. Functional enrichment analysis showed that these DEGs were mainly enriched in cancer-related pathways and were primarily involved in biological processes (BP) such as cell and mitotic division. From the Hub gene screening, PLK1 was further identified as the Hub gene associated with TNBC. Cell and animal experiments indicated that PLK1 promotes the proliferation, invasion, migration, and clone formation of breast cancer cells.

CONCLUSIONS

Gene chip combined with bioinformatics methods can effectively analyze the DEGs related to the occurrence and development of breast cancer, and the screening of PLK1 can provide theoretical guidance for further research on the molecular mechanism of breast cancer and the screening of molecular markers.

摘要

背景

乳腺癌是女性中最常见的恶性肿瘤。三阴性乳腺癌(TNBC)是指一种特殊的亚型,其雌激素(ER)、孕激素(PR)和人表皮生长因子受体2(HER-2)表达缺失。在本研究中,使用了多种生物信息学分析工具来筛选与三阴性乳腺癌发生发展相关的枢纽基因,并分析其生物学功能。

方法

选择基因表达综合数据库(GEO)中的乳腺癌微阵列数据GSE62931作为研究对象。使用生物信息学工具筛选差异表达基因(DEGs),并构建DEGs的蛋白质-蛋白质相互作用(PPI)网络。同时筛选枢纽基因。利用基因本体论(GO)知识库和京都基因与基因组百科全书(KEGG)进行生物富集分析。使用基因表达谱交互式分析(GEPIA)在线工具验证筛选基因的表达及患者生存情况。使用细胞计数试剂盒8(CCK-8)、transwell迁移试验、划痕试验和克隆形成试验验证polo样激酶1(PLK1)对乳腺癌细胞增殖、侵袭、迁移和干性的影响。建立乳腺癌皮下异种移植动物模型,以评估PLK1对乳腺癌增殖的影响。

结果

通过GSE62931微阵列数据共筛选出824个DEGs;其中405个上调,419个下调。功能富集分析表明,这些DEGs主要富集在癌症相关途径中,主要参与细胞和有丝分裂等生物过程(BP)。通过枢纽基因筛选,进一步确定PLK1为与TNBC相关的枢纽基因。细胞和动物实验表明,PLK1促进乳腺癌细胞的增殖、侵袭、迁移和克隆形成。

结论

基因芯片结合生物信息学方法可有效分析与乳腺癌发生发展相关的DEGs,PLK1的筛选可为进一步研究乳腺癌分子机制及筛选分子标志物提供理论指导。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0796/7812170/d083dc0c4fa6/atm-08-24-1637-f1.jpg

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