CAJ | 학술논문

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.
목적 관찰특이성소간우RNA(siRNA)대간암세포거서세포이동억제인자(MIF)기인표체적억제작용.방법 지질체방법장siRNA전염간암세포PLC、Hep3B.정량RT-PCR、Western blot검측MIF mRNA화단백、MAPK신호분자적표체.새서람(MTY)비색법、체외세포침습실험검측세포증식화대중조기저막(matrigel)천투능력.결과 100 nmol/L적MIF siRNA사MIF mR-NA표체하조81.3%화89.1%,단백수평강저69.50%、72.31%,여대조조비교기차이유통계학의의(P균＜0.01).세포증식솔하강16.79%화47.14%(P균＜0.05).천투matrigel적세포수위51.00±11.27화18.56±4.72,여대조조비교차이유통계학의의(P균＜0.05).린산화MAPK하조.결론 MIF siRNA유효억제MIF표체급간암세포증식화천이,가능부분통과억제MAPK린산화기작용.
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.