结直肠癌（CRC）作为一种致命的癌症，是全世界最常见的癌症之一。尽管结直肠癌的标准治疗已得到充分研究和确立，但患者的长期生存率仍然很低，死亡率仍然很高。因此，需要越来越多有效的治疗方案。为了评价贝伐单抗、组蛋白去甲基酶抑制剂IOX1或它们的组合治疗结直肠癌的疗效，我们检测了IOX1、贝伐单抗以及IOX1联合贝伐单抗对结直肠癌细胞系的细胞活性、增殖和迁移的影响。 HCT116、RKO 和 CT26（通过 CCK8）、集落形成测定、伤口愈合测定和 Transwell 测定。通过流式细胞术检查了药物单独以及组合对结直肠癌细胞系凋亡的影响，并通过凋亡相关蛋白的蛋白质印迹进一步验证。在动物模型中检查了单独或联合治疗对结直肠癌细胞的抗肿瘤作用。小鼠皮下注射CT26细胞，药物治疗后通过IHC检测肿瘤组织的生长和免疫浸润情况。我们发现IOX1能够有效抑制CRC细胞的活性，对CRC细胞的增殖和迁移有显着的抑制作用。 IOX1处理结直肠癌细胞后，细胞凋亡率呈剂量依赖性增加，凋亡相关蛋白的表达也随之变化。与贝伐单抗的进一步联合显示，与单独使用IOX1或贝伐单抗相比，该组合对CRC细胞的增殖、迁移和凋亡具有更显着的影响。体内实验发现，单独用药和联合用药均能抑制小鼠肿瘤的生长，但联合用药抑制效果最为明显。联合治疗显着抑制肿瘤异种移植模型中增殖标志物（Ki67）的表达，并增加抗原特异性CD4、CD8 T细胞生长的含量，并在联合治疗中检测到与T细胞细胞毒性相关的粒酶B（GZMB） 。免疫测定抑制相关 PD-1 的表达并降低。抗癌药物贝伐珠单抗和组蛋白去甲基化酶抑制剂IOX1可能通过调节细胞凋亡来抑制结肠癌细胞的生长。联合治疗对肿瘤生长的抑制作用可能部分是通过T淋巴细胞上调浸润介导的肿瘤免疫来实现的。 IOX1和贝伐珠单抗的组合产生了显着的协同效应。本研究旨在为CRC联合治疗提供新方向。版权所有©2024。Elsevier B.V.出版。

Colorectal cancer (CRC), as a fatal cancer, is one of the most common cancers worldwide. Although the standard treatment for colorectal cancer is well researched and established, long-term patient survival remains poor, and mortality remains high. Therefore, more and more effective treatment options are needed. To evaluate the efficacy of bevacizumab, the histone demethylase inhibitor IOX1, or their combination for the treatment of colorectal cancer, we examined the effects of IOX1, bevacizumab, and IOX1 combined with bevacizumab on cell activity, proliferation, and migration of colorectal cancer cell lines HCT116, RKO, and CT26 by CCK8, colony formation assay, wound healing assay, and transwell assay. The effects of the drugs alone as well as in combination on apoptosis in colorectal cancer cell lines were examined by flow cytometry and further validated by Western blotting for apoptosis-related proteins. The antitumor effects of treatment alone or in combination on colorectal cancer cells were examined in animal models. Mice were injected subcutaneously with CT26 cells and the growth and immune infiltration in tumor tissues were detected by IHC after drug treatment. We found that IOX1 could effectively inhibit the activity of CRC cells and had a significant inhibitory effect on the proliferation and migration of CRC cells. The apoptosis rate increased in a dose-dependent manner after IOX1 treatment on colorectal cancer cells, and the expression of apoptosis-related proteins changed accordingly. Further combination with bevacizumab revealed that the combination had a more significant effect on the proliferation, migration, and apoptosis of CRC cells than either IOX1 or bevacizumab alone. In vivo experiments have found that both alone and combination drugs can inhibit the growth of mouse tumors, but the effect of combination inhibition is the most obvious. Combination therapy significantly inhibited the expression of proliferative marker (Ki67) in tumor xenograft models, and increased content of antigen-specific CD4+, CD8+T cell growth, and granzymeB (GZMB), which is associated with T cell cytotoxicity, was detected in combination therapy. Immunoassays suppressed the expression of relevant PD-1 and decreased. The anticancer drug bevacizumab and the histone demethylase inhibitor IOX1 may inhibit colon cancer cell growth by regulating apoptosis. The inhibitory effect of combination therapy on tumor growth may be achieved, in part, through upregulation of infiltration-mediated tumor immunity by T lymphocytes. The combination of IOX1 and bevacizumab produced significant synergistic effects. This study aims to provide a new direction for CRC combination therapy.Copyright © 2024. Published by Elsevier B.V.