中华肝脏病杂志
中華肝髒病雜誌
중화간장병잡지
CHINESE JOURNAL OF HEPATOLOGY
2009年
2期
102-106
,共5页
夏金堂%李雯%伍兆锋%赵杰%王花%李瑜元
夏金堂%李雯%伍兆鋒%趙傑%王花%李瑜元
하금당%리문%오조봉%조걸%왕화%리유원
癌,肝细胞%RNA干扰%周期素依赖激酶抑制剂p27%巨噬细胞移动抑制因子
癌,肝細胞%RNA榦擾%週期素依賴激酶抑製劑p27%巨噬細胞移動抑製因子
암,간세포%RNA간우%주기소의뢰격매억제제p27%거서세포이동억제인자
Carcinoma,hepatocellular%RNA interference%Cyclin-dependent kinase inhibitor p27%Macrophage migration inhibitor factor
目的 观察巨噬细胞移动抑制因子(MIF)和细胞周期调控因子p27在肝细胞癌中表达的相互关系,探讨小干扰RNA(siRNA)沉默MIF基因对肝癌细胞p27表达的影响.方法 免疫组织化学法和荧光定量PCR法检测MIF、p27的蛋白和mRNA在肝癌及其癌旁组织中的表达情况.化学合成MIF siRNA和对照siRNA,脂质体法转染肝癌细胞PLC和Hep3B,荧光定量PCR法检测MIF和p27 mRNA在实验组及对照组中的表达情况.根据不同资料分别采用X2检验、Logistic回归分析或单因素方差分析.结果 MIF蛋白及其mRNA在肝癌组织中过表达,在癌旁组织中低表达;p27蛋白及其mRNA在癌组织中低表达,在癌旁组织中高表达.Logistic回归分析提示MIF为肝癌发生的危险因素,p27为保护因素.MIF mRNA在肝癌细胞株中过表达(F=61.036,P＜0.01),p27 mRNA在正常肝细胞L02中高表达(F=529.853,P＜0.01).经MIFsiRNA转染后,MIF mRNA在PLC及Hep3B中的表达水平降低,并且呈剂量依赖关系(F值分别为320.1和201.2,P值均＜0.01);p27 mRNA伴随MIF mRNA的降低而增加(F值分别为419.4和459.9,P值均＜0.01).结论 MIF在肝细胞癌中过表达,MIF siRNA能特异性抑制其在肝癌细胞中的表达;MIF可能参与了p27基因表达的调控.
目的 觀察巨噬細胞移動抑製因子(MIF)和細胞週期調控因子p27在肝細胞癌中錶達的相互關繫,探討小榦擾RNA(siRNA)沉默MIF基因對肝癌細胞p27錶達的影響.方法 免疫組織化學法和熒光定量PCR法檢測MIF、p27的蛋白和mRNA在肝癌及其癌徬組織中的錶達情況.化學閤成MIF siRNA和對照siRNA,脂質體法轉染肝癌細胞PLC和Hep3B,熒光定量PCR法檢測MIF和p27 mRNA在實驗組及對照組中的錶達情況.根據不同資料分彆採用X2檢驗、Logistic迴歸分析或單因素方差分析.結果 MIF蛋白及其mRNA在肝癌組織中過錶達,在癌徬組織中低錶達;p27蛋白及其mRNA在癌組織中低錶達,在癌徬組織中高錶達.Logistic迴歸分析提示MIF為肝癌髮生的危險因素,p27為保護因素.MIF mRNA在肝癌細胞株中過錶達(F=61.036,P＜0.01),p27 mRNA在正常肝細胞L02中高錶達(F=529.853,P＜0.01).經MIFsiRNA轉染後,MIF mRNA在PLC及Hep3B中的錶達水平降低,併且呈劑量依賴關繫(F值分彆為320.1和201.2,P值均＜0.01);p27 mRNA伴隨MIF mRNA的降低而增加(F值分彆為419.4和459.9,P值均＜0.01).結論 MIF在肝細胞癌中過錶達,MIF siRNA能特異性抑製其在肝癌細胞中的錶達;MIF可能參與瞭p27基因錶達的調控.
목적 관찰거서세포이동억제인자(MIF)화세포주기조공인자p27재간세포암중표체적상호관계,탐토소간우RNA(siRNA)침묵MIF기인대간암세포p27표체적영향.방법 면역조직화학법화형광정량PCR법검측MIF、p27적단백화mRNA재간암급기암방조직중적표체정황.화학합성MIF siRNA화대조siRNA,지질체법전염간암세포PLC화Hep3B,형광정량PCR법검측MIF화p27 mRNA재실험조급대조조중적표체정황.근거불동자료분별채용X2검험、Logistic회귀분석혹단인소방차분석.결과 MIF단백급기mRNA재간암조직중과표체,재암방조직중저표체;p27단백급기mRNA재암조직중저표체,재암방조직중고표체.Logistic회귀분석제시MIF위간암발생적위험인소,p27위보호인소.MIF mRNA재간암세포주중과표체(F=61.036,P＜0.01),p27 mRNA재정상간세포L02중고표체(F=529.853,P＜0.01).경MIFsiRNA전염후,MIF mRNA재PLC급Hep3B중적표체수평강저,병차정제량의뢰관계(F치분별위320.1화201.2,P치균＜0.01);p27 mRNA반수MIF mRNA적강저이증가(F치분별위419.4화459.9,P치균＜0.01).결론 MIF재간세포암중과표체,MIF siRNA능특이성억제기재간암세포중적표체;MIF가능삼여료p27기인표체적조공.
Objective To observe the expression of macrophage migration inhibition factor(MIF)and p27 in hepatocellular carcinoma tissue,and to investigate the effect of MIF on the expression of p27 in hepatocellular carcinoma(HCC)cells.Methods Immunohistochemistry and quantitative RT-PCR were performed to detect the expression of MIF and p27 in HCC tissues and peri-tumor tissues.Specific small interfering RNA(siRNA)targeting MIF gene was chemically synthesized and then transfected at the concentration of 50 nmol/L and 100 nmol/L into PLC cells and Hep3B cells.The mRNA levels of MIF and p27 after MIF siRNA treatment were quantified by real-time RT-PCR.Results MIF protein and mRNA were overexpressed in the HCC tumor tissues compared to these in the peri-tumor tissues (P＜0.01).The expression of p27 protein and mRNA was significantly lower in the HCC tumor tissues compared to these in the peri-tumor tissues(P＜0.01).Compared to nodal liver cell line L-02,HCC cell lines expressed higher level of MIF (F=61.036,P＜0.01)and lower level of p27(F=529.853,P＜0.01).In MIF siRNA treated PLC and Hep3B cells,the MIF mRNA was decreased in a dose-dependent manner(F=320.1,P＜0.01;F=201.2,P＜0.01).The p27 mRNA was significantly up-regulated in MIF siRNA treated PLC and Hep3B cells compared to control siRNA transfected cells(F=419.4,P＜0.01;F=459.9,P＜0.01).Conelusions MIF iS overexpressed in HCC tumor tissues,and the expression of p27 is repressed by MIF.
