Chang In-Youb, Boo Hye-Jin, Hyun Jin Won, Yoon Sang-Pil
Department of Anatomy, College of Medicine, Chosun University, Gwangju 61452, Republic of Korea.
Jeju Research Center for Natural Medicine, Jeju National University, Jeju 63243, Republic of Korea.
Oncol Lett. 2025 Mar 24;29(5):245. doi: 10.3892/ol.2025.14991. eCollection 2025 May.
Although E-cadherin is known as a tumor suppressor via its effects on cell to cell adhesion, the effects of E-cadherin on malignant transformation have not yet been thoroughly investigated. In the present study, after malignant transformation was induced by spheroid formation in a fetal bovine serum-supplemented environment, the effects of soluble E-cadherin on the spheroidogenesis of colorectal cancer cells were investigated. E-cadherin knock-out (KO) was performed in HCT116 cells, targeting exon 3 of the CDH1 gene. A cell viability assay was performed to determine the proliferation and viability of wild type and CDH1 KO HCT116 cells after treatment with anticancer drugs. Spheroidogenesis was compared with or without exogenous E-cadherin, antibody against the ectodomain of E-cadherin (DECMA-1) and PD98059 treatment. In addition, morphometry, immunocytochemistry and western blotting were performed. Soluble E-cadherin in culture media was measured using an enzyme-linked immunosorbent assay. Firstly, CDH1 KO was confirmed by western blotting. Notably, the proliferation and viability of cells following treatment with 5-fluorouracil, epidermal growth factor receptor inhibitor and src kinase inhibitor were similar between the cell lines. Exogenous E-cadherin or DECMA-1 treatment did not affect spheroidogenesis, although long-term maintenance was slightly disturbed in CDH1 KO spheroids compared with that in wild type spheroids. In addition, E-cadherin was increased in spheroid culture as compared with that in conventional culture. Soluble E-cadherin was increased in a time-dependent manner, particularly in wild type HCT116 cells. PD98059 inhibited ERK activation and enhanced E-cadherin expression in conventional culture without affecting spheroidogenesis. These results suggested that soluble E-cadherin may be considered as a biomarker for colorectal cancer, although exogenous E-cadherin might not have a further role in malignant transformation.
尽管E-钙黏蛋白通过其对细胞间黏附的作用而被认为是一种肿瘤抑制因子，但E-钙黏蛋白对恶性转化的影响尚未得到充分研究。在本研究中，在添加胎牛血清的环境中通过球体形成诱导恶性转化后，研究了可溶性E-钙黏蛋白对结肠癌细胞球体形成的影响。在HCT116细胞中进行E-钙黏蛋白基因敲除（KO），靶向CDH1基因的外显子3。在用抗癌药物处理后，进行细胞活力测定以确定野生型和CDH1基因敲除的HCT116细胞的增殖和活力。比较有无外源性E-钙黏蛋白、抗E-钙黏蛋白胞外域抗体（DECMA-1）和PD98059处理时的球体形成情况。此外，还进行了形态测量、免疫细胞化学和蛋白质免疫印迹分析。使用酶联免疫吸附测定法测量培养基中的可溶性E-钙黏蛋白。首先，通过蛋白质免疫印迹分析确认了CDH1基因敲除。值得注意的是，在用5-氟尿嘧啶、表皮生长因子受体抑制剂和src激酶抑制剂处理后，细胞系之间细胞的增殖和活力相似。外源性E-钙黏蛋白或DECMA-1处理不影响球体形成，尽管与野生型球体相比，CDH1基因敲除球体的长期维持略有干扰。此外，与传统培养相比，球体培养中E-钙黏蛋白增加。可溶性E-钙黏蛋白以时间依赖性方式增加，特别是在野生型HCT116细胞中。PD98059在传统培养中抑制ERK激活并增强E-钙黏蛋白表达，而不影响球体形成。这些结果表明，可溶性E-钙黏蛋白可被视为结直肠癌的生物标志物，尽管外源性E-钙黏蛋白在恶性转化中可能没有进一步作用。
