Yu Qing, Qu Chunhua, Liang Jinliang, Chen Peiqi, Zhang Kaiying, Zhang Yanji, Zhang Yuening, Li Zherui, Liu Shaojun, Yang Zhaoshou, Sun Hongyan, Yang Anli
Department of Clinical Laboratory, Shenzhen Kangning Hospital, Shenzhen, 518000, P. R. China.
Department of Clinical Laboratory, Maternal & Child Health Hospital of Foshan, Foshan, 528000, P.R. China.
Clin Proteomics. 2024 Jul 15;21(1):50. doi: 10.1186/s12014-024-09502-8.
Estrogen receptor-positive (ER+) breast cancer accounts for two-thirds of all breast cancers, and its early and late recurrences still threaten patients' long-term survival and quality of life. Finding candidate tumor antigens and potential therapeutic targets is critical to addressing these unmet needs.
The isobaric tags for relative and absolute quantitation (iTRAQ) proteomic analysis was employed to identify the differentially expressed proteins (DEPs) between ER + breast cancer and corresponding adjacent normal tissue. Candidate DEPs were screened by bioinformatic analyses, and their expression was confirmed by immunohistochemical (IHC) staining and western blot. A series of in vitro experiments, including wound healing assay, colony formation, and cell cycle assay, were performed to reveal the functions of selected DEPs. Additionally, their clinical significances were further analyzed.
A total of 369 DEPs (fold change ≥ 2.0 or ≤ 0.66, P < 0.05) were discovered. Compared with normal tissue, 358 proteins were up-regulated and 11 proteins were down-regulated in ER + breast cancer. GO and KEGG enrichment analysis showed that DEPs were closely associated with RNA regulation and metabolic pathways. STRING analysis found ESF1 and MIPEP were the hub genes in breast cancer, whose increased expressions were verified by the IHC staining and western blot. Knocking down ESF1 and MIPEP inhibited colony formation and increased cell apoptosis. Besides, knocking down ESF1 inhibited wound healing but not MIPEP. In addition, ESF1 and MIPEP expression were negatively associated with patient prognosis.
The upregulation of ESF1 and MIPEP promoted ER + breast cancer proliferation, which might provide novel targets for the development of new therapies.
雌激素受体阳性（ER+）乳腺癌占所有乳腺癌的三分之二，其早期和晚期复发仍然威胁着患者的长期生存和生活质量。寻找候选肿瘤抗原和潜在治疗靶点对于满足这些未满足的需求至关重要。
采用相对和绝对定量等压标签（iTRAQ）蛋白质组学分析来鉴定ER+乳腺癌与相应癌旁正常组织之间的差异表达蛋白（DEP）。通过生物信息学分析筛选候选DEP，并通过免疫组织化学（IHC）染色和蛋白质免疫印迹法确认其表达。进行了一系列体外实验，包括伤口愈合试验、集落形成和细胞周期试验，以揭示所选DEP的功能。此外，进一步分析了它们的临床意义。
共发现369个DEP（倍数变化≥2.0或≤0.66，P<0.05）。与正常组织相比，ER+乳腺癌中有358种蛋白质上调，11种蛋白质下调。基因本体（GO）和京都基因与基因组百科全书（KEGG）富集分析表明，DEP与RNA调控和代谢途径密切相关。蛋白质相互作用网络分析（STRING）发现ESF1和MIPEP是乳腺癌中的枢纽基因，其表达增加通过IHC染色和蛋白质免疫印迹法得到验证。敲低ESF1和MIPEP可抑制集落形成并增加细胞凋亡。此外，敲低ESF1可抑制伤口愈合，但敲低MIPEP则无此作用。另外，ESF1和MIPEP的表达与患者预后呈负相关。
ESF1和MIPEP的上调促进了ER+乳腺癌的增殖，这可能为开发新疗法提供新的靶点。
