Department of Medical Biochemistry, Faculty of Medicine, Akdeniz University, 07070, Antalya, Turkey.
Department of Medical Microbiology, Faculty of Medicine, Akdeniz University, 07070, Antalya, Turkey.
Naunyn Schmiedebergs Arch Pharmacol. 2021 Nov;394(11):2259-2272. doi: 10.1007/s00210-021-02139-6. Epub 2021 Aug 26.
Cyclooxygenase-2 (COX-2) is expressed in a variety of human colorectal cancer cells and can contribute to carcinogenesis. This study aimed to investigate the effect of diclofenac (DCF), a selective COX-2 inhibitor, on cell adhesion molecules and apoptosis in human colon adenocarcinoma cells. Levels of homing cell adhesion molecule (H-CAM, CD44), intercellular adhesion molecule-1 (ICAM-1, CD54), vascular cell adhesion molecule-1 (VCAM-1, CD106), and epithelial cell adhesion molecule (EpCAM, CD326) were evaluated in cancer cells overexpressing (HT29) or not expressing (HCT116) COX-2. Cell viability was determined by MTT assay, COX-2 protein levels and activity were assessed by immunofluorescence and fluorometric analysis, respectively. Endogenous levels of polyunsaturated fatty acids (PUFAs) were measured by liquid chromatography-tandem mass spectrometry (LC-MS/MS) while expression of cell adhesion molecules was analyzed by flow cytometry. Annexin V-FITC/propidium iodide-labelling and fluorometric caspase-3 activity measurements were carried out to determine apoptosis. Flow cytometry analysis revealed that the percentage of CD44 and ICAM-1 staining in HCT116 cells was significantly lower compared to HT29 cells. In HT29 cells, phorbol 12-myristate 13-acetate (PMA) induced COX-2 expression and increased CD44 and ICAM-1 levels were down-regulated by diclofenac. Stimulation of COX-2 activity in HT29 cells via PMA significantly decreased diclofenac associated increase in PUFA levels. Treatment with both diclofenac and PMA significantly increased the number of apoptotic cells and caspase-3 activity in colon adenocarcinoma cells compared to control groups. In conclusion, diclofenac's effect to retard colorectal tumor growth and metastasis occurs in COX-2 overexpressing colon cancer cells by increased apoptosis and decreased expression of CD44 and ICAM-1.
环氧化酶-2（COX-2）在多种人类结直肠癌细胞中表达，可促进癌变。本研究旨在探讨选择性 COX-2 抑制剂双氯芬酸（DCF）对人结肠腺癌细胞黏附分子和凋亡的影响。评估高表达（HT29）或不表达（HCT116）COX-2 的癌细胞中归巢细胞黏附分子（H-CAM，CD44）、细胞间黏附分子-1（ICAM-1，CD54）、血管细胞黏附分子-1（VCAM-1，CD106）和上皮细胞黏附分子（EpCAM，CD326）的水平。通过 MTT 测定法测定细胞活力，通过免疫荧光和荧光分析分别评估 COX-2 蛋白水平和活性。通过液相色谱-串联质谱（LC-MS/MS）测量多不饱和脂肪酸（PUFA）的内源性水平，通过流式细胞术分析细胞黏附分子的表达。通过 Annexin V-FITC/碘化丙啶标记和荧光酶活性测定法测定 caspase-3 活性来确定凋亡。流式细胞术分析显示，与 HT29 细胞相比，HCT116 细胞中 CD44 和 ICAM-1 染色的百分比明显较低。在 HT29 细胞中，佛波醇 12-肉豆蔻酸 13-乙酸酯（PMA）诱导 COX-2 表达，并下调双氯芬酸诱导的 CD44 和 ICAM-1 水平增加。PMA 刺激 HT29 细胞 COX-2 活性显著降低了双氯芬酸相关的 PUFA 水平增加。与对照组相比，双氯芬酸和 PMA 联合处理显著增加了结肠腺癌细胞的凋亡细胞数量和 caspase-3 活性。总之，双氯芬酸通过增加凋亡和降低 CD44 和 ICAM-1 的表达来减缓结直肠肿瘤生长和转移的作用发生在 COX-2 过表达的结肠癌细胞中。
