Muhammad Saud Tabish, Yiwei Li, Na Zhao, Yichen Hong, Jiao Shu, Chenbo Yang, Zexin Sun, Xiaonan Liu, Miaomiao Sun, Kuisheng Chen
{"title":"Relationship between CCM3 expression and angiogenesis in esophageal squamous cell carcinoma and its possible mechanism.","authors":"Muhammad Saud Tabish, Yiwei Li, Na Zhao, Yichen Hong, Jiao Shu, Chenbo Yang, Zexin Sun, Xiaonan Liu, Miaomiao Sun, Kuisheng Chen","doi":"10.1007/s10147-026-03132-w","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong>Angiogenesis is essential for esophageal squamous cell carcinoma (ESCC) progression, yet clinically actionable regulators remain limited. We investigated the expression, functional role, and mechanism of CCM3 in ESCC and its relationship with the HIF-1α/VEGFA angiogenic axis.</p><p><strong>Methods: </strong>CCM3, HIF-1α, VEGFA and microvessel density (MVD, CD31) were assessed by immunohistochemistry in 53 paired ESCC and adjacent non-cancerous tissues. CCM3 knockdown was optimized using four siRNAs in KYSE-70 and KYSE270 cells. CCM3 siRNA 1064-A was selected as the most effective and optimal knockdown achieved at 8 nM, 48 h. Tumor-conditioned HUVECs (referred to as tumor-conditioned endothelial cells, TECs) were generated using conditioned media from knockdown cells; TEC proliferation (CCK8), migration (scratch), invasion (Transwell) and tube formation were evaluated in vitro. A subcutaneous xenograft model using KYSE-70 cells was employed in vivo, followed by intratumoral silencing of CCM3 to assess tumour development, proliferation (Ki-67), and microvessel density (CD31).</p><p><strong>Results: </strong>High endogenous CCM3 expression was significantly upregulated in ESCC tissues compared to adjacent non-cancerous tissues and correlated with clinicopathological features like depth of invasion, lymph node metastasis and advanced stage. CCM3 expression positively correlated with HIF-1α and VEGFA. CCM3 knockdown siRNA 1064-A attenuates TEC proliferation, migration, invasion and tube formation in vitro. In vivo, CCM3 knockdown reduced tumor growth, tumor weight, Ki-67, CD31 staining and levels of HIF-1α and VEGFA.</p><p><strong>Conclusions: </strong>High endogenous CCM3 expression associates with aggressive ESCC clinicopathological features and angiogenesis markers. CCM3 knockdown inhibits tumor-conditioned endothelial function, xenograft growth, intratumoral vascularization, and HIF-1α/VEGFA expression, warranting further mechanistic investigation.</p>","PeriodicalId":13869,"journal":{"name":"International Journal of Clinical Oncology","volume":" ","pages":"2191-2212"},"PeriodicalIF":3.0000,"publicationDate":"2026-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"International Journal of Clinical Oncology","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1007/s10147-026-03132-w","RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2026/7/8 0:00:00","PubModel":"Epub","JCR":"Q3","JCRName":"ONCOLOGY","Score":null,"Total":0}
引用次数: 0
Abstract
Background: Angiogenesis is essential for esophageal squamous cell carcinoma (ESCC) progression, yet clinically actionable regulators remain limited. We investigated the expression, functional role, and mechanism of CCM3 in ESCC and its relationship with the HIF-1α/VEGFA angiogenic axis.
Methods: CCM3, HIF-1α, VEGFA and microvessel density (MVD, CD31) were assessed by immunohistochemistry in 53 paired ESCC and adjacent non-cancerous tissues. CCM3 knockdown was optimized using four siRNAs in KYSE-70 and KYSE270 cells. CCM3 siRNA 1064-A was selected as the most effective and optimal knockdown achieved at 8 nM, 48 h. Tumor-conditioned HUVECs (referred to as tumor-conditioned endothelial cells, TECs) were generated using conditioned media from knockdown cells; TEC proliferation (CCK8), migration (scratch), invasion (Transwell) and tube formation were evaluated in vitro. A subcutaneous xenograft model using KYSE-70 cells was employed in vivo, followed by intratumoral silencing of CCM3 to assess tumour development, proliferation (Ki-67), and microvessel density (CD31).
Results: High endogenous CCM3 expression was significantly upregulated in ESCC tissues compared to adjacent non-cancerous tissues and correlated with clinicopathological features like depth of invasion, lymph node metastasis and advanced stage. CCM3 expression positively correlated with HIF-1α and VEGFA. CCM3 knockdown siRNA 1064-A attenuates TEC proliferation, migration, invasion and tube formation in vitro. In vivo, CCM3 knockdown reduced tumor growth, tumor weight, Ki-67, CD31 staining and levels of HIF-1α and VEGFA.
Conclusions: High endogenous CCM3 expression associates with aggressive ESCC clinicopathological features and angiogenesis markers. CCM3 knockdown inhibits tumor-conditioned endothelial function, xenograft growth, intratumoral vascularization, and HIF-1α/VEGFA expression, warranting further mechanistic investigation.
期刊介绍：
The International Journal of Clinical Oncology (IJCO) welcomes original research papers on all aspects of clinical oncology that report the results of novel and timely investigations. Reports on clinical trials are encouraged. Experimental studies will also be accepted if they have obvious relevance to clinical oncology. Membership in the Japan Society of Clinical Oncology is not a prerequisite for submission to the journal. Papers are received on the understanding that: their contents have not been published in whole or in part elsewhere; that they are subject to peer review by at least two referees and the Editors, and to editorial revision of the language and contents; and that the Editors are responsible for their acceptance, rejection, and order of publication.
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