背景:含有JumonjiC(JmjC)结构域的蛋白质参与了肿瘤发生和癌症进展。然而,这种效应的潜在机制仍然知之甚少。我们的目的是研究Jumonji和富含AT的相互作用域2(JARID2)-JmjC家族蛋白-在乳腺癌中的作用,以及它与肥胖的潜在联系。
方法:免疫组织化学,癌症基因组图谱,基因表达综合,和其他数据库用于分析JARID2在乳腺癌细胞中的表达。生长曲线,5-乙炔基-2-脱氧尿苷(EdU),菌落形成,和细胞侵袭实验用于检测JARID2是否影响乳腺癌细胞增殖和侵袭。NOD/SCID小鼠中基于球化的实验和异种肿瘤移植用于检查JARID2与乳腺癌干性之间的关联。RNA测序,京都基因和基因组百科全书,和基因集富集分析用于鉴定JARID2参与的细胞过程。进行免疫亲和纯化和银染质谱以搜索可能与JARID2相互作用的蛋白质。使用共免疫沉淀和谷胱甘肽S-转移酶(GST)下拉实验进一步验证了结果。使用染色质免疫沉淀(ChIP)测序,我们寻找JARID2和转移相关蛋白1(MTA1)共同调控的靶基因;结果通过ChIP-PCR验证,定量ChIP(qChIP)和ChIP-reChIP测定。共培养实验用于探索乳腺癌细胞与脂肪细胞之间的相互作用。
结果:在这项研究中,我们发现JARID2在包括乳腺癌在内的多种癌症中高表达.JARID2促进糖酵解,脂质代谢,扩散,入侵,和乳腺癌细胞的干性。此外,JARID2与核小体重塑和脱乙酰酶(NuRD)复合物物理相互作用,转录抑制一系列肿瘤抑制基因,如BRCA2DNA修复相关(BRCA2),RB转录辅抑制因子1(RB1),和肌醇多磷酸-4-磷酸酶IIB型(INPP4B)。此外,在乳腺癌微环境中,JARID2的表达由肥胖相关的脂肪因子瘦素通过Janus激酶2/信号转导和转录激活因子3(JAK2/STAT3)途径调节。各种在线数据库的分析也表明JARID2/MTA1与乳腺癌的不良预后相关。
结论:我们的数据表明JARID2促进乳腺肿瘤发生和发展,确认JARID2作为癌症治疗的靶标。
BACKGROUND: Proteins containing the Jumonji C (JmjC) domain participated in tumorigenesis and cancer progression. However, the mechanisms underlying this effect are still poorly understood. Our objective was to investigate the role of Jumonji and the AT-rich interaction domain-containing 2 (
JARID2) - a JmjC family protein - in breast cancer, as well as its latent association with obesity.
METHODS: Immunohistochemistry, The Cancer Genome Atlas, Gene Expression Omnibus, and other databases were used to analyze the expression of JARID2 in breast cancer cells. Growth curve, 5-ethynyl-2-deoxyuridine (EdU), colony formation, and cell invasion experiments were used to detect whether JARID2 affected breast cancer cell proliferation and invasion. Spheroidization-based experiments and xenotumor transplantation in NOD/SCID mice were used to examine the association between JARID2 and breast cancer stemness. RNA-sequencing, Kyoto Encyclopedia of Genes and Genomes, and Gene Set Enrichment Analysis were used to identify the cell processes in which JARID2 participates. Immunoaffinity purification and silver staining mass spectrometry were conducted to search for proteins that might interact with JARID2. The results were further verified using co-immunoprecipitation and glutathione S-transferase (GST) pull-down experiments. Using chromatin immunoprecipitation (ChIP) sequencing, we sought the target genes that JARID2 and metastasis-associated protein 1 (MTA1) jointly regulated; the results were validated by ChIP-PCR, quantitative ChIP (qChIP) and ChIP-reChIP assays. A coculture experiment was used to explore the interactions between breast cancer cells and adipocytes.
RESULTS: In this study, we found that JARID2 was highly expressed in multiple types of cancer including breast cancer.
JARID2 promoted glycolysis, lipid metabolism, proliferation, invasion, and stemness of breast cancer cells. Furthermore,
JARID2 physically interacted with the nucleosome remodeling and deacetylase (NuRD) complex, transcriptionally repressing a series of tumor suppressor genes such as BRCA2 DNA repair associated (BRCA2), RB transcriptional corepressor 1 (RB1), and inositol polyphosphate-4-phosphatase type II B (INPP4B). Additionally, JARID2 expression was regulated by the obesity-associated adipokine leptin via Janus kinase 2/signal transducer and activator of transcription 3 (JAK2/STAT3) pathway in the breast cancer microenvironment. Analysis of various online databases also indicated that JARID2/MTA1 was associated with a poor prognosis of breast cancer.
CONCLUSIONS: Our data indicated that
JARID2 promoted breast tumorigenesis and development, confirming
JARID2 as a target for cancer treatment.