VEGF介导微囊藻毒素-LR促进结直肠癌细胞DLD-1侵袭迁移的研究

VEGF expression in microcystin-LR promotion of invasion and migration in DLD-1 cells

目的:研究微囊藻毒素-LR是否调节结直肠癌细胞VEGF的表达,进而影响肿瘤细胞的侵袭与迁移。方法:结直肠癌细胞DLD-1经过不用浓度微囊藻毒素-LR处理后,Transwell实验分析其侵袭、迁移能力的改变。实时荧光定量PCR(qPCR)、Western blot以及ELISA实验检测DLD-1细胞中VEGF mRNA和蛋白表达水平的变化。结果:Transwell实验结果表明,与对照组比较,微囊藻毒素-LR处理组细胞侵袭、迁移能力明显提高,当微囊藻毒素-LR的浓度达到12.5 nmol/L时,侵袭、迁移能力分别提高了2.1和2.0倍;qPCR、Western blot结果显示微囊藻毒素-LR上调了结直肠癌细胞DLD-1中VEGF mRNA以及蛋白的表达水平,当微囊藻毒素-LR的浓度达到12.5和25 nmol/L时,DLD-1细胞中VEGF mRNA和蛋白水平分别达到最大值,上升了1.7和2.8倍;同时ELISA实验也检测到在细胞培养液中DLD-1细胞分泌的VEGF提高了1.5倍。结论:微囊藻毒素-LR可以显著上调DLD-1细胞中VEGF的表达,进而介导肿瘤细胞发生侵袭与迁移。

OBJECTIVE：This study investigated whether microcystin-LR regulated the expression of VEGF and promoted invasion and migration in colorectal DLD-1 cells.METHODS：Transwell experiments were used to analyze the invasion and migration in colorectal cancer DLD-1 cells treated with different concentrations of microcystin-LR.Expression of VEGF in treated cells was evaluated by real-time quantitative PCR（qPCR）,Western blot and ELISA.RESULTS：That the abilities of invasion and migration improved significantly in microcystin-LR-treated groups,compared with those in the control group.When the concentration of microcystin-LR reached 12.5nmol/L,invasion and migration were increased to 2.1 and 2.0 folds,respectively.The qPCR and Western blot results suggested that microcystin-LR upregulated the expression level of VEGF mRNA and protein in DLD-1 cells.When microcystin-LR concentration reached 12.5 and 25 nmol/L,the expression levels of the VEGF protein and mRNA in DLD-1 cells reached a maximum of 1.7 and 2.8 folds increases,respectively.Mean while,the significant 1.5-foldelevated VEGF production in the culture medium was also detected.CONCLUSION：Microcystin-LR upregulation of VEGF was involved in promoting invasion and migration in colorectal cancer cells DLD-1.