大肠癌细胞及相关成纤维细胞18F-脱氧葡萄糖摄取能力研究

2-deoxy-2-18 F-fluoro-D-glucose uptake capability of human colorectal cancer cells and colorectal cancer associated fibroblasts

目的 观察人大肠癌细胞及其相关成纤维细胞（CCAFs）共培养前后的18F-脱氧葡萄糖（18 F-FDG）摄取能力的变化,分析CCAFs的糖代谢特征及其对肿瘤代谢的影响.方法 分别改变反应细胞数（1 ×104、5×104、1 ×105、3×105、5×105）、18F-FDG反应时间（40、60、80、100、120 min）、反应放射性活度（1.85、3.70、7.40、14.80、29.60 kBq）以及初始葡萄糖浓度（0、2.78、5.55、11.10 mmol/L）,筛选出CCAFs与18F-FDG结合的最佳条件,并测定CCAFs-大肠癌细胞共培养4d前后Caco-2及HCT116两种大肠癌细胞和CCAFs的18F-FDG摄取率.结果 CCAFs结合18F-FDG的最佳条件为：细胞数为5×105个/瓶,18F-FDG放射性活度为3.70 kBq,反应时间为100 min,葡萄糖浓度为0mmol/L,细胞结合率为（42.80 ±4.47）％.线性回归方程为：Y=（2.661＋0.463E4X1-0.195X2-2.412X3）×100％,F=51.140,P＜0.05;共培养前后CCAFs的18F-FDG结合率差异无统计学意义（P＞0.05）,Caco-2细胞共培养前后结合率分别为（28.650 0±4.760 0）％、（18.821 5±1.5100）％ （P＜0.05）;HCT116细胞共培养前后18F-FDG结合率分别为（24.974 3±2.8300）％、（17.7202 ±2.6200）％,共培养的CCAFs与肿瘤细胞比较,差异均有统计学意义（P＜0.05）.结论 CCAFs的18F-FDG摄取能力明显大于大肠癌细胞,而且CCAFs与大肠癌细胞共培养,可明显抑制后者对18F-FDG的摄取而对CCAFs的摄取能力无明显影响,提示CCAFs可能是大肠癌组织中葡萄糖摄取的重要细胞成分.

Objective To investigate the capability of 2-deoxy-2-18F-fluoro-D-glucose （18F-FDG） uptake by human colorectal cancer cells and colorectal cancer associated fibroblasts （CCAFs） before and after co-culture of these two kinds of cells and to explore the characteristics of glucose metabolism of CCAFs.Methods The binding conditions of CCAFs and 18F-FDG were opitimized regarding to cells count （1 ×104,5 ×104,1 ×105,3 ×105,5 ×105）,reaction time （40,60,80,100,120 min）,radioactivity of 18F-FDG （1.85,3.70,7.40,14.80,29.60 kBq） and initial glucose concentration （0,2.78,5.55,11.10 mmol/L）.The uptake of 18F-FDG by Caco-2,HCT116 and CCAFs was determined before and after co-culture of CCAFs-cancer cells.Results The optimal conditions of 18F-FDG binding were as follows：cell counts 5 × 105/wefl,reaction time 100 min,radioactivity of 18F-FDG 3.70 kBq and initial glucose concentration 0 mmol/L,and the 18F-FDG cells binding ratio of CCAFs was （42.80 ±4.47）%.Before and after co-culture,the 18F-FDG binding ratio of Caco-2 was （28.6500± 4.7600）%,and （18.821 5± 1.5100）% （P〈 0.05）,and that of HCT116 was （24.9743± 2.8300）%,and （17.720 2 ± 2.620 0） % （P 〈 0.05）.No significant change in 1s F-FDG binding by CCAFs was observed.Conclusion The uptake capability of 18F-FDG by CCAFs was sinifiantly higher than colorectal cancer cells.Co-culture with CCAFs siginificantly suppressed the uptake of colorectal cancer cells.CCAFs may be a very important component in the enhanced uptake of glucose in colorectal cancer.