Department of General Surgery, Beijing Tiantan Hospital, Capital Medical University, Beijing 100070, P.R. China.
J Microbiol Biotechnol. 2024 Nov 28;34(11):2211-2222. doi: 10.4014/jmb.2407.07008. Epub 2024 Sep 20.
The current pathological and physiological evaluation system for colorectal cancer (CRC) is limited; thus, effective biological targets to diagnose and treat this disease are urgently needed. In this study, we used qRT-PCR for detecting mRNA levels of genes. The levels of protein were identified by western blot, immunohistochemistry, and immunofluorescence assays. In addition, functional experiments were used to evaluate the role of cytoskeleton associated protein (CKAP) 2 in CRC cells and human umbilical vein endothelial cells (HUVECs). Bioinformatics analysis was employed to predict the binding relationship of CKAP2 and TFDP1, which was confirmed through dual luciferase reporter assay and immunoprecipitation assay. Furthermore, we injected human colorectal carcinoma HCT116 cells into mice flanks, and we injected Luciferase-labeled HCT116 cells into mice tail vein. HE staining was used to detect tumor nodules. As a result, high CKAP2 expression was found in CRC cells and tissues. CKAP2 silencing reduced CRC cell migration, invasion, proliferation, and epithelial-mesenchymal transition. Moreover, CKAP2 expression was positively associated with M2 macrophage levels. CKAP2 promoted protein expression of CD86, CD206, IL-1β, and CCL17. Moreover, CKAP2 promoted the proliferation of HUVECs and angiogenesis via affecting the tumor microenvironment (TME). We also found that CKAP2 could interact with TFDP1. The inhibitory impacts of TFDP1 downregulation on CRC cell' proliferation, migration, and invasion were reversed via CKAP2 overexpression. In vivo silencing of CKAP2 repressed tumor growth and metastasis. Overall, CKAP2 was positively regulated by TFDP1, which promoted tumorigenesis and metastasis in CRC.
目前用于结直肠癌（CRC）的病理和生理评估系统有限；因此，迫切需要有效的生物靶标来诊断和治疗这种疾病。在这项研究中，我们使用 qRT-PCR 检测基因的 mRNA 水平。通过 Western blot、免疫组织化学和免疫荧光测定来鉴定蛋白质水平。此外，还使用功能实验来评估细胞骨架相关蛋白（CKAP）2 在 CRC 细胞和人脐静脉内皮细胞（HUVEC）中的作用。生物信息学分析用于预测 CKAP2 和 TFDP1 的结合关系，通过双荧光素酶报告基因检测和免疫沉淀检测证实了这一点。此外，我们将人结直肠癌细胞 HCT116 注入小鼠侧腹，并将 Luciferase 标记的 HCT116 细胞注入小鼠尾静脉。通过 HE 染色检测肿瘤结节。结果发现，CRC 细胞和组织中 CKAP2 表达较高。CKAP2 沉默降低了 CRC 细胞的迁移、侵袭、增殖和上皮间质转化。此外，CKAP2 的表达与 M2 巨噬细胞水平呈正相关。CKAP2 促进 CD86、CD206、IL-1β 和 CCL17 的蛋白表达。此外，CKAP2 通过影响肿瘤微环境（TME）促进 HUVEC 增殖和血管生成。我们还发现 CKAP2 可以与 TFDP1 相互作用。通过 CKAP2 过表达，TFDP1 下调对 CRC 细胞增殖、迁移和侵袭的抑制作用被逆转。体内沉默 CKAP2 抑制肿瘤生长和转移。总之，TFDP1 正向调节 CKAP2，促进 CRC 中的肿瘤发生和转移。
