{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,1]],"date-time":"2026-05-01T19:36:13Z","timestamp":1777664173565,"version":"3.51.4"},"reference-count":36,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2025,10,20]],"date-time":"2025-10-20T00:00:00Z","timestamp":1760918400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Joint Funds for the Innovation of Science and Technology, Fujian Province","award":["2024Y9193"],"award-info":[{"award-number":["2024Y9193"]}]},{"name":"Joint Funds for the Innovation of Science and Technology, Fujian Province","award":["2020Y9131"],"award-info":[{"award-number":["2020Y9131"]}]},{"name":"Fujian Provincial Finance Project","award":["2020B010"],"award-info":[{"award-number":["2020B010"]}]},{"DOI":"10.13039\/501100003392","name":"Natural Science Foundation of Fujian Province","doi-asserted-by":"crossref","award":["2021J01703"],"award-info":[{"award-number":["2021J01703"]}],"id":[{"id":"10.13039\/501100003392","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["CIMB"],"abstract":"<jats:p>Background: Ductal carcinoma in situ (DCIS) is a precursor to breast cancer. The mechanisms by which the stroma of DCIS affects disease progression remain elusive. Thus, the aim of this study is to identify key stroma genes that affect DCIS progression and to define high-risk DCIS cases. Method: Gene expression matrix files from the Gene Expression Omnibus (GEO) database were selected to identify candidate genes associated with the stromal transition from DCIS to invasive ductal carcinoma (IDC). An integrative approach was employed to identify and functionally characterize driver genes of DCIS progression. In vitro experiments were performed to validate the role of these genes. Results: We identified 13 differentially expressed genes (DEGs), of which 5 were selected as candidate drivers. Gene set enrichment analysis (GSEA) revealed the biological functions of RAMP2 and ADM2, while in vitro functional assays demonstrated that ADM2 knockdown and RAMP2 overexpression in breast cancer cell lines significantly suppressed cellular proliferation and invasion. Conclusion: This study identified and validated the roles and functions of ADM2 and RAMP2 and revealed their function as key driver genes in the progression of ductal carcinoma in situ (DCIS). Collectively, our findings elucidate critical genetic mechanisms underlying DCIS progression and provide novel insights for the development of personalized therapeutic strategies.<\/jats:p>","DOI":"10.3390\/cimb47100864","type":"journal-article","created":{"date-parts":[[2025,10,20]],"date-time":"2025-10-20T13:54:41Z","timestamp":1760968481000},"page":"864","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Integrating Multiple Methods to Validate Key Genes Driving the Progression of Breast Ductal Carcinoma In Situ"],"prefix":"10.3390","volume":"47","author":[{"given":"Minjie","family":"Zhong","sequence":"first","affiliation":[{"name":"Department of Thyroid and Breast Surgery, The First Affiliated Hospital of Fujian Medical University, Fuzhou 350005, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shengkai","family":"Zheng","sequence":"additional","affiliation":[{"name":"Department of Thyroid and Breast Surgery, The First Affiliated Hospital of Fujian Medical University, Fuzhou 350005, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yahui","family":"Wen","sequence":"additional","affiliation":[{"name":"Department of Thyroid and Breast Surgery, The First Affiliated Hospital of Fujian Medical University, Fuzhou 350005, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Juansi","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Thyroid and Breast Surgery, The First Affiliated Hospital of Fujian Medical University, Fuzhou 350005, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jiahui","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Thyroid and Breast Surgery, The First Affiliated Hospital of Fujian Medical University, Fuzhou 350005, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hanwei","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Thyroid and Breast Surgery, The First Affiliated Hospital of Fujian Medical University, Fuzhou 350005, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Caiqin","family":"Mo","sequence":"additional","affiliation":[{"name":"Department of Thyroid and Breast Surgery, The First Affiliated Hospital of Fujian Medical University, Fuzhou 350005, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sunwang","family":"Xu","sequence":"additional","affiliation":[{"name":"Department of Thyroid and Breast Surgery, The First Affiliated Hospital of Fujian Medical University, Fuzhou 350005, China"},{"name":"National Regional Medical Center, Department of Thyroid and Breast Surgery, Binhai Campus of the First Affiliated Hospital, Fujian Medical University, Fuzhou 350212, China"},{"name":"Fujian Provincial Key Laboratory, Precision Medicine for Cancer, Fuzhou 350000, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xiangjin","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Thyroid and Breast Surgery, The First Affiliated Hospital of Fujian Medical University, Fuzhou 350005, China"},{"name":"National Regional Medical Center, Department of Thyroid and Breast Surgery, Binhai Campus of the First Affiliated Hospital, Fujian Medical University, Fuzhou 350212, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,10,20]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"859","DOI":"10.1016\/j.molonc.2013.07.005","article-title":"Progression from ductal carcinoma in situ to invasive breast cancer: Revisited","volume":"7","author":"Cowell","year":"2013","journal-title":"Mol. Oncol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"487","DOI":"10.1038\/s41575-020-0300-1","article-title":"Pancreatic cancer stroma: An update on therapeutic targeting strategies","volume":"17","author":"Hosein","year":"2020","journal-title":"Nat. Rev. Gastroenterol. Hepatol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"209","DOI":"10.1097\/JTO.0b013e3181f8a1bd","article-title":"The role of tumor stroma in cancer progression and prognosis: Emphasis on carcinoma-associated fibroblasts and non-small cell lung cancer","volume":"6","author":"Bremnes","year":"2011","journal-title":"J. Thorac. Oncol."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Low, J.-Y., and Laiho, M. (2022). Caveolae-Associated Molecules, Tumor Stroma, and Cancer Drug Resistance: Current Findings and Future Perspectives. Cancers, 14.","DOI":"10.3390\/cancers14030589"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"118","DOI":"10.1186\/s13058-021-01494-9","article-title":"The role of stromal immune microenvironment in the progression of ductal carcinoma in situ (DCIS) to invasive breast cancer","volume":"23","author":"Olszewski","year":"2021","journal-title":"Breast Cancer Res."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"983","DOI":"10.1093\/jnci\/djy234","article-title":"Impact of Epithelial-Stromal Interactions on Peritumoral Fibroblasts in Ductal Carcinoma in Situ","volume":"111","author":"Strell","year":"2019","journal-title":"J. Natl. Cancer Inst."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"580","DOI":"10.4161\/cc.27379","article-title":"JNK1 stress signaling is hyper-activated in high breast density and the tumor stroma: Connecting fibrosis, inflammation, and stemness for cancer prevention","volume":"13","author":"Lisanti","year":"2014","journal-title":"Cell Cycle"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"462","DOI":"10.5858\/arpa.2012-0078-RA","article-title":"Ductal Carcinoma In Situ of the Breast: Current Concepts and Future Directions","volume":"137","author":"Siziopikou","year":"2013","journal-title":"Arch. Pathol. Lab. Med."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"453","DOI":"10.1038\/nmeth.3337","article-title":"Robust enumeration of cell subsets from tissue expression profiles","volume":"12","author":"Newman","year":"2015","journal-title":"Nat. Methods"},{"key":"ref_10","first-page":"100625","article-title":"Integrated analysis of public datasets for the discovery and validation of survival-associated genes in solid tumors","volume":"5","year":"2024","journal-title":"Innovation"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"W98","DOI":"10.1093\/nar\/gkx247","article-title":"GEPIA: A web server for cancer and normal gene expression profiling and interactive analyses","volume":"45","author":"Tang","year":"2017","journal-title":"Nucleic Acids Res."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Zhang, H., Zhao, Y., Liu, X., Fu, L., Gu, F., and Ma, Y. (2021). High Expression of Complement Component C7 Indicates Poor Prognosis of Breast Cancer and Is Insensitive to Taxane-Anthracycline Chemotherapy. Front. Oncol., 11.","DOI":"10.3389\/fonc.2021.724250"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1456","DOI":"10.1038\/s41388-021-02172-y","article-title":"STAT3 induces breast cancer growth via ANGPTL4, MMP13 and STC1 secretion by cancer associated fibroblasts","volume":"41","author":"Avalle","year":"2022","journal-title":"Oncogene"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"690","DOI":"10.1038\/s41419-021-03960-w","article-title":"Overexpression of PSMC2 promotes the tumorigenesis and development of human breast cancer via regulating plasminogen activator urokinase (PLAU)","volume":"12","author":"Wang","year":"2021","journal-title":"Cell Death Dis."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1038\/s41422-019-0257-1","article-title":"ILF3 is a substrate of SPOP for regulating serine biosynthesis in colorectal cancer","volume":"30","author":"Li","year":"2020","journal-title":"Cell Res."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"372","DOI":"10.1158\/0008-5472.CAN-17-1912","article-title":"SHMT2 Desuccinylation by SIRT5 Drives Cancer Cell Proliferation","volume":"78","author":"Yang","year":"2018","journal-title":"Cancer Res."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"194","DOI":"10.1186\/s12957-023-03078-7","article-title":"Identification of DDIT4 as a potential prognostic marker associated with chemotherapeutic and immunotherapeutic response in triple-negative breast cancer","volume":"21","author":"Chen","year":"2023","journal-title":"World J. Surg. Oncol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1318","DOI":"10.1038\/s41590-023-01529-7","article-title":"The GPCR-G\u03b1(s)-PKA signaling axis promotes T cell dysfunction and cancer immunotherapy failure","volume":"24","author":"Wu","year":"2023","journal-title":"Nat. Immunol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1038\/nrc2069","article-title":"G-protein-coupled receptors and cancer","volume":"7","author":"Dorsam","year":"2007","journal-title":"Nat. Rev. Cancer"},{"key":"ref_20","first-page":"5440","article-title":"Development of a metabolism-related signature for predicting prognosis, immune infiltration and immunotherapy response in breast cancer","volume":"12","author":"Li","year":"2022","journal-title":"Am. J. Cancer Res."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"222","DOI":"10.1038\/nature10138","article-title":"CCL2 recruits inflammatory monocytes to facilitate breast-tumour metastasis","volume":"475","author":"Qian","year":"2011","journal-title":"Nature"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"850","DOI":"10.1038\/s41588-022-01082-3","article-title":"Genomic analysis defines clonal relationships of ductal carcinoma in situ and recurrent invasive breast cancer","volume":"54","author":"Lips","year":"2022","journal-title":"Nat. Genet."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"4063","DOI":"10.1016\/j.csbj.2020.11.040","article-title":"Derivation of a nuclear heterogeneity image index to grade DCIS","volume":"18","author":"Hayward","year":"2020","journal-title":"Comput. Struct. Biotechnol. J."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1038\/s41392-024-01779-3","article-title":"Progression from ductal carcinoma in situ to invasive breast cancer: Molecular features and clinical significance","volume":"9","author":"Wang","year":"2024","journal-title":"Signal Transduct. Target. Ther."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"114","DOI":"10.1186\/s12943-021-01412-9","article-title":"CircMYH9 drives colorectal cancer growth by regulating serine metabolism and redox homeostasis in a p53-dependent manner","volume":"20","author":"Liu","year":"2021","journal-title":"Mol. Cancer"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"645","DOI":"10.1016\/j.tcb.2017.05.001","article-title":"Serine and Functional Metabolites in Cancer","volume":"27","author":"Newman","year":"2017","journal-title":"Trends Cell Biol."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Wu, Q., Li, J., Sun, S., Chen, X., Zhang, H., Li, B., and Sun, S. (2017). YAP\/TAZ-mediated activation of serine metabolism and methylation regulation is critical for LKB1-deficient breast cancer progression. Biosci. Rep., 37.","DOI":"10.1042\/BSR20171072"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"197","DOI":"10.1126\/science.abf1730","article-title":"The G protein signaling regulator RGS3 enhances the GTPase activity of KRAS","volume":"374","author":"Li","year":"2021","journal-title":"Science"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"e2117004119","DOI":"10.1073\/pnas.2117004119","article-title":"Progesterone activates GPR126 to promote breast cancer development via the Gi pathway","volume":"119","author":"An","year":"2022","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"e2205395","DOI":"10.1002\/advs.202205395","article-title":"Combined Single-Cell and Spatial Transcriptomics Reveal the Metabolic Evolvement of Breast Cancer during Early Dissemination","volume":"10","author":"Liu","year":"2023","journal-title":"Adv. Sci."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"235","DOI":"10.1002\/cncr.34519","article-title":"Immune scoring model based on immune cell infiltration to predict prognosis in diffuse large B-cell lymphoma","volume":"129","author":"Yang","year":"2023","journal-title":"Cancer"},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Kotwal, A., Gustafson, M.P., Bornschlegl, S., Dietz, A.B., Delivanis, D., and Ryder, M. (2023). Circulating immunophenotypes are potentially prognostic in follicular cell-derived thyroid cancer. Front. Immunol., 14.","DOI":"10.3389\/fimmu.2023.1325343"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"588","DOI":"10.1016\/j.ccell.2019.02.009","article-title":"Human Tumor-Associated Macrophage and Monocyte Transcriptional Landscapes Reveal Cancer-Specific Reprogramming, Biomarkers, and Therapeutic Targets","volume":"35","author":"Cassetta","year":"2019","journal-title":"Cancer Cell"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"2950","DOI":"10.7150\/thno.29617","article-title":"CD137 promotes bone metastasis of breast cancer by enhancing the migration and osteoclast differentiation of monocytes\/macrophages","volume":"9","author":"Jiang","year":"2019","journal-title":"Theranostics"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"e1346","DOI":"10.1002\/ctm2.1346","article-title":"Single-cell transcriptome sequencing of B-cell heterogeneity and tertiary lymphoid structure predicts breast cancer prognosis and neoadjuvant therapy efficacy","volume":"13","author":"Wang","year":"2023","journal-title":"Clin. Transl. Med."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"15840","DOI":"10.1074\/jbc.RA118.005062","article-title":"Structure-function analyses reveal a triple \u03b2-turn receptor-bound conformation of adrenomedullin 2\/intermedin and enable peptide antagonist design","volume":"293","author":"Roehrkasse","year":"2018","journal-title":"J. Biol. Chem."}],"container-title":["Current Issues in Molecular Biology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1467-3045\/47\/10\/864\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,22]],"date-time":"2025-10-22T04:32:51Z","timestamp":1761107571000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1467-3045\/47\/10\/864"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,10,20]]},"references-count":36,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2025,10]]}},"alternative-id":["cimb47100864"],"URL":"https:\/\/doi.org\/10.3390\/cimb47100864","relation":{},"ISSN":["1467-3045"],"issn-type":[{"value":"1467-3045","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,10,20]]}}}