RNA干扰对肝癌细胞巨噬细胞移动抑制因子基因表达的抑制作用

目的 观察特异性小干扰RNA(siRNA)对肝癌细胞巨噬细胞移动抑制因子(MIF)基因表达的抑制作用.方法 脂质体方法将siRNA转染肝癌细胞PLC、Hep3B.定量RT-PCR、Western blot检测MIF mRNA和蛋白、MAPK信号分子的表达.噻唑蓝(MTY)比色法、体外细胞侵袭实验检测细胞增殖和对重组基底膜(matrigel)穿透能力.结果 100 nmol/L的MIF siRNA使MIF mR-NA表达下调81.3%和89.1%,蛋白水平降低69.50%、72.31%,与对照组比较其差异有统计学意义(P均＜0.01).细胞增殖率下降16.79%和47.14%(P均＜0.05).穿透matrigel的细胞数为51.00±11.27和18.56±4.72,与对照组比较差异有统计学意义(P均＜0.05).磷酸化MAPK下调.结论 MIF siRNA有效抑制MIF表达及肝癌细胞增殖和迁移,可能部分通过抑制MAPK磷酸化起作用.

Inhibitory effect of siRNA on the expression of macrophage migration inhibitory factor gene in hepatocyte carcinoma cells

Objective To investigate the inhibitory effects of specific small interfering RNA(siRNA)on the expression of migration inhibitory factor(MIF)gene in hepatocyte carcinoma cells.Methods Double strain siRNAs which were synthesized by chemical methods were transfected into PLC and Hep3B cells with liposome method.The expression of MIF mRNA and protein and MAPK levels were detected by quantitative RT-PCR,Western blot and immtmofluorescent staining.Tumor cell proliferation was tested by MTT metllod,and cell migration measured by counting the cell number of trans-well.Results The expression of MIF mRNA was decreased by 81.3%and 89.1%and MIF protein levels were decreased by 69.50%and 72.31%respectively in PLC and HeD3B cells when compared with control groups 24 h after MIF siRNA transfection(all P＜0.01).MIF immunostaining was obviously reduced.Cell proliferation in MIF siRNA groups was significanfly decreased by 16.79%and 47.14%respectively as compared with controls(both P＜0.05);The cell number of migration was 51.00±11.27(PLC)and 18.56±4.72(Hep3B)respectively per well after 48 h trans-well growth,which were significantly decreased as compared with controls(both P＜0.05).Phosphorylated MAPK level was decreased both in PLC and Hep3B as compared with controls.Conclusion Specific MIF siRNA call down-regulate the MIF gene expression and inhibit tumor cell growth and migration,which may be related with MAP kinase signal pathway.