Department of Oncology, Affiliated Hospital of Chengde Medical University, Chengde, China.
J Gene Med. 2020 Aug;22(8):e3195. doi: 10.1002/jgm.3195. Epub 2020 May 7.
The present study aimed to investigate the expression, function and clinical implication of zinc finger protein 750 (ZNF750) in colonic cancer and explore the mechanism of its dysregulation.
The expression of ZNF750 in 76 pairs of colonic cancer tissues was determined using immunohistochemistry. The expression of ZNF750 in colonic cancer cells was detected using western blotting. The correlation between the expression level of ZNF750 and clinicopathological parameters in patients with colonic cancer was analyzed using a chi-squared test. CCK-8 and colony formation assays were used to monitor cell proliferation. Additionally, flow cytometry was used to detect apoptosis of cells; scratch healing and Transwell assays were conducted to evaluate the migration and invasion of cells. Ultimately, the binding relationship between miR-17-5p and ZNF750 was validated using western blotting, a real-time polymerase chaub reaction and a dual-luciferase reporter gene assay.
The expression level of ZNF750 in colonic cancer tissues, as well as colonic cancer cell lines, was significantly down-regulated. Low expression of ZNF750 was associated with larger tumor size and poor tumor differentiation. The over-expression of ZNF750 inhibited the proliferation, motility and invasion but promoted the apoptosis of colonic cancer cells. After the cells were transfected with miR-17-5p mimics, the expression of ZNF750 at both mRNA and protein levels was markedly decreased, whereas the expression of ZNF750 was markedly increased after transfection of miR-17-5p inhibitors. MiR-17-5p could suppresses the malignant biological behaviors via negatively regulating ZNF750.
ZNF750 is negatively regulated by miR-17-5p and inhibits the progression of colonic cancer.
本研究旨在探讨锌指蛋白 750（ZNF750）在结肠癌中的表达、功能及其临床意义，并探讨其失调的机制。
采用免疫组织化学法检测 76 对结肠癌组织中 ZNF750 的表达，采用 Western blot 法检测结肠癌细胞中 ZNF750 的表达，采用卡方检验分析 ZNF750 表达水平与结肠癌患者临床病理参数的相关性。采用 CCK-8 及集落形成实验检测细胞增殖情况，流式细胞术检测细胞凋亡情况，划痕愈合实验及 Transwell 实验检测细胞迁移及侵袭能力。采用 Western blot、实时荧光定量聚合酶链式反应及双荧光素酶报告基因实验验证 miR-17-5p 与 ZNF750 的结合关系。
ZNF750 在结肠癌组织及结肠癌细胞系中的表达水平均显著下调，ZNF750 低表达与肿瘤体积较大、肿瘤分化较差有关。过表达 ZNF750 抑制了结肠癌细胞的增殖、迁移及侵袭能力，促进了细胞凋亡。转染 miR-17-5p 模拟物后，ZNF750 的 mRNA 和蛋白表达水平均显著降低，而转染 miR-17-5p 抑制剂后，ZNF750 的表达水平显著升高。miR-17-5p 可通过负调控 ZNF750 抑制结肠癌的恶性生物学行为。
ZNF750 受 miR-17-5p 负调控，抑制结肠癌的进展。
