转染反义MIF对人胃癌细胞的影响

摘要

Objective To explore the effect of antisense macrophage migration inhibitory factor (MIF) on hu-man 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 pcD-NA3.1-sh-MIF (control group). The qRT-PCR and Western blot analysis were employed for detecting transfection effi-ciency. 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 re-sults showed that the expression of MIF mRNA in pcDNA3.1-Anti MIF group was significantly lower than that in con-trol 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 as-say 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 dif-ferences were all statistically significant (P<0.01). Conclusion Antisense MIF can inhibit the proliferation and inva-sion of human gastric cancer MGC-803 cells, as well as induce apoptosis.%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-AntiMIF)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、AnnexinV-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 mRNA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,pcDNA3.1vAntiMIF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-AntiMIF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-AntiMIF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;AnnexinV-FITC和PI染色法结果显示,pcDNA3.1-AntiMIF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.