转染反义MIF对人胃癌细胞的影响  

Effects of antisense macrophage migration inhibitory factor in human gastric cancer cells

作　　者：张雪梅 孙延亮[2] 杨扬 兰辉 王喜梅 

机构地区：[1]湖南医药学院,湖南怀化418000 [2]怀化市第一人民医院,湖南怀化418000 [3]湖南医药学院第一附属医院,湖南怀化418000

出　　处：《海南医学》2017年第9期1377-1380,共4页Hainan Medical Journal

基　　金：湖南省教育厅科学研究项目(编号:13C741)

摘　　要：目的探讨反义巨噬细胞移动抑制因子(MIF)对人胃癌MGC-803细胞的影响。方法将胃癌MGC-803细胞分为pcDNA3.1-Anti MIF组与空白对照组。pcDNA3.1-Anti MIF组采用转染技术将MIF反义RNA真核表达质粒(pcDNA3.1-Anti MIF)转入MGC-803细胞;空白对照组转染pcDNA3.1-sh-MIF质粒。qRT-PCR与蛋白质印迹法检测转染效率;采用MTT法、侵袭实验、Annexin V-FITC和PI染色法分别检测反义MIF对MGC-803细胞增殖、侵袭、凋亡的影响。结果 qRT-PCR结果显示,pcDNA3.1-Anti MIF组MIF m RNA表达量(2.086±0.248)较空白对照组(6.992±0.342)明显下调;Western blot显示,pcDNA3.1v Anti MIF组MIF蛋白表达水平量(0.361±0.043)较空白对照组(1.171±0.091)明显下调;MTT实验结果显示,pcDNA3.1-Anti MIF组MGC-803细胞的OD值(0.436±0.017)较空白对照组(0.563±0.019)明显下降;侵袭实验结果显示,pcDNA3.1-Anti MIF组穿过基质胶的MGC-803细胞数(73.67±8.54)较空白对照组(137.30±11.91)明显减少;Annexin V-FITC和PI染色法结果显示,pcDNA3.1-Anti MIF组MGC-803细胞的凋亡率(21.61±4.62)%较空白对照组(7.67±0.63)%明显增加。以上各项指标比较差异均具有显著统计学意义(P<0.01)。结论反义MIF能抑制人胃癌MGC-803细胞的增殖与侵袭,并能诱导凋亡。Objective To explore the effect of antisense macrophage migration inhibitory factor（MIF） on human gastric cancer MGC-803 cells. Methods Human gastric cancer MGC-803 cells were assigned into two groups to receive transfection of antisense MIF plasmid pcDNA3.1-Anti MIF（pcDNA3.1-Anti MIF group） and pcDNA3.1-shMIF（control group）. The qRT-PCR and Western blot analysis were employed for detecting transfection efficiency. The ability of proliferation and invasion, as well as apoptosis rate of MGC-803 cells regulated by antisense MIF were evaluated by MTT, Transwell invasion assays, and Annexin V/propidium iodide staining. Results qRT-PCR results showed that the expression of MIF m RNA in pcDNA3.1-Anti MIF group was significantly lower than that in control group,（2.086±0.248） vs（6.992±0.342）. Western blot analysis results showed that the expression of MIF protein in pcDNA3.1-Anti MIF group was significantly lower than that in control group,（0.361±0.043） vs（1.171±0.091）. MTT assay showed that the OD value of MGC-803 cells in the pcDNA3.1-Anti MIF group was significantly lower than that in control group,（0.436 ± 0.017） vs（0.563 ± 0.019）. Transwell invasion assays results showed that the number of cells through the matris glue in pcDNA3.1-Anti MIF group was significantly lower than that in the control group,（73.67 ± 8.54） vs（137.30 ± 11.91）. Annexin V/propidium iodide staining showed that the apoptosis rate of MGC-803 cells in the pcDNA3.1-Anti MIF group was significantly higher than that in control group,（21.61±4.62）% vs（7.67±0.63）%. The differences were all statistically significant（P〈0.01）. Conclusion Antisense MIF can inhibit the proliferation and invasion of human gastric cancer MGC-803 cells, as well as induce apoptosis.

关 键 词：巨噬细胞移动抑制因子 胃癌 增殖 侵袭 凋亡 

分 类 号：R735.2[医药卫生—肿瘤]