中华普通外科杂志
中華普通外科雜誌
중화보통외과잡지
CHINESE JOURNAL OF GENERAL SURGERY
2009年
5期
398-401
,共4页
夏金堂%伍兆锋%李雯%赖越元%赵杰%徐晨%王花%滕元%李瑜元
夏金堂%伍兆鋒%李雯%賴越元%趙傑%徐晨%王花%滕元%李瑜元
하금당%오조봉%리문%뢰월원%조걸%서신%왕화%등원%리유원
癌,肝细胞%巨噬细胞游走抑制因子%RNA干扰%细胞周期蛋白D1
癌,肝細胞%巨噬細胞遊走抑製因子%RNA榦擾%細胞週期蛋白D1
암,간세포%거서세포유주억제인자%RNA간우%세포주기단백D1
Carcinoma,hepatocellular%Macrophage migration inhibitory factors%RNA interference%Cyclin D1
目的 探讨巨噬细胞游走抑制因子(macrophage migration inhibitory factor,MIF)和细胞周期蛋白D1(cyclin D1)在原发性肝细胞癌(hepatocellular carcinoma,HCC)组织中的表达及两者在肝癌发病机制及细胞周期调控中的关系.方法 应用定量PCR及Westem blot技术检测MIF和Cyclin D1在肝癌组织和癌旁组织的表达.化学合成MIF siRNA,将PLC细胞和HepG2细胞分成对照组、MIF siRNA 50 nmol/L干预组、MIF siRNA 1130 nmol/L干预组,脂质体法转染肝癌细胞PLC和HepG2,定量PCR技术和Western blot检测MIF siRNA干扰后MIF和Cyclin D1 mRNA和蛋白的表达变化.结果 MIF和Cyclin D1蛋白在肝癌组织中的相对表达量为0.825±0.13和0.843±0.104;MIF和Cyclin D1 mRNA在肝癌组织中的相对表达量为癌旁组织的(7.31±1.85)倍和(4.27±1.05)倍,与癌旁组织相比,差异均具有统计学意义(FMIF=15.5,P<0.01;fCyclin D1=87.5,P<0.01).应用小RNA干扰技术转染PLC和HepG2肝癌细胞后MIF mRNA分别下调71.2%±7.2%、87.4%±2.9%、74.3%±8.9%、88.4%±4.6%(FPLC=315.5,P<0.01;FHHepG2=201.2,P<0.01);MIF蛋白分别下调为0.33±0.03、0.11±0.02、0.81±0.08、0.36±0.02,并呈剂量依赖关系(FPLC=43.9,P<0.01;FHepG2:133.4,P<0.01).伴随MIF mRNA和蛋白的表达F调Cyclin D1 mRNA分别下调68.2%±3%、78.1%±1.4%、65.8%±4.7%、77.3%±2.6%(FPLC=1569,P<0.01;FHepG2=480.4,P<0.01);Cyclin D1蛋白下调0.28±0.06、0.15±0.03、0.44±0.04、0.13±0.02,亦呈剂量依赖关系,与对照组相比差异有统计学意义(P<0.01),两干预组相比差异有统计学意义(FPLC=35.5,P<0.01;FHepG2=114.7,P<0.01).结论 MIF和Cyclin D1在肝细胞癌中过表达,MIF可能在转录水平调控Cyclin D1的表达,并促使肿瘤细胞通过细胞周期检测点,继续增值和分化,导致肿瘤的形成.
目的 探討巨噬細胞遊走抑製因子(macrophage migration inhibitory factor,MIF)和細胞週期蛋白D1(cyclin D1)在原髮性肝細胞癌(hepatocellular carcinoma,HCC)組織中的錶達及兩者在肝癌髮病機製及細胞週期調控中的關繫.方法 應用定量PCR及Westem blot技術檢測MIF和Cyclin D1在肝癌組織和癌徬組織的錶達.化學閤成MIF siRNA,將PLC細胞和HepG2細胞分成對照組、MIF siRNA 50 nmol/L榦預組、MIF siRNA 1130 nmol/L榦預組,脂質體法轉染肝癌細胞PLC和HepG2,定量PCR技術和Western blot檢測MIF siRNA榦擾後MIF和Cyclin D1 mRNA和蛋白的錶達變化.結果 MIF和Cyclin D1蛋白在肝癌組織中的相對錶達量為0.825±0.13和0.843±0.104;MIF和Cyclin D1 mRNA在肝癌組織中的相對錶達量為癌徬組織的(7.31±1.85)倍和(4.27±1.05)倍,與癌徬組織相比,差異均具有統計學意義(FMIF=15.5,P<0.01;fCyclin D1=87.5,P<0.01).應用小RNA榦擾技術轉染PLC和HepG2肝癌細胞後MIF mRNA分彆下調71.2%±7.2%、87.4%±2.9%、74.3%±8.9%、88.4%±4.6%(FPLC=315.5,P<0.01;FHHepG2=201.2,P<0.01);MIF蛋白分彆下調為0.33±0.03、0.11±0.02、0.81±0.08、0.36±0.02,併呈劑量依賴關繫(FPLC=43.9,P<0.01;FHepG2:133.4,P<0.01).伴隨MIF mRNA和蛋白的錶達F調Cyclin D1 mRNA分彆下調68.2%±3%、78.1%±1.4%、65.8%±4.7%、77.3%±2.6%(FPLC=1569,P<0.01;FHepG2=480.4,P<0.01);Cyclin D1蛋白下調0.28±0.06、0.15±0.03、0.44±0.04、0.13±0.02,亦呈劑量依賴關繫,與對照組相比差異有統計學意義(P<0.01),兩榦預組相比差異有統計學意義(FPLC=35.5,P<0.01;FHepG2=114.7,P<0.01).結論 MIF和Cyclin D1在肝細胞癌中過錶達,MIF可能在轉錄水平調控Cyclin D1的錶達,併促使腫瘤細胞通過細胞週期檢測點,繼續增值和分化,導緻腫瘤的形成.
목적 탐토거서세포유주억제인자(macrophage migration inhibitory factor,MIF)화세포주기단백D1(cyclin D1)재원발성간세포암(hepatocellular carcinoma,HCC)조직중적표체급량자재간암발병궤제급세포주기조공중적관계.방법 응용정량PCR급Westem blot기술검측MIF화Cyclin D1재간암조직화암방조직적표체.화학합성MIF siRNA,장PLC세포화HepG2세포분성대조조、MIF siRNA 50 nmol/L간예조、MIF siRNA 1130 nmol/L간예조,지질체법전염간암세포PLC화HepG2,정량PCR기술화Western blot검측MIF siRNA간우후MIF화Cyclin D1 mRNA화단백적표체변화.결과 MIF화Cyclin D1단백재간암조직중적상대표체량위0.825±0.13화0.843±0.104;MIF화Cyclin D1 mRNA재간암조직중적상대표체량위암방조직적(7.31±1.85)배화(4.27±1.05)배,여암방조직상비,차이균구유통계학의의(FMIF=15.5,P<0.01;fCyclin D1=87.5,P<0.01).응용소RNA간우기술전염PLC화HepG2간암세포후MIF mRNA분별하조71.2%±7.2%、87.4%±2.9%、74.3%±8.9%、88.4%±4.6%(FPLC=315.5,P<0.01;FHHepG2=201.2,P<0.01);MIF단백분별하조위0.33±0.03、0.11±0.02、0.81±0.08、0.36±0.02,병정제량의뢰관계(FPLC=43.9,P<0.01;FHepG2:133.4,P<0.01).반수MIF mRNA화단백적표체F조Cyclin D1 mRNA분별하조68.2%±3%、78.1%±1.4%、65.8%±4.7%、77.3%±2.6%(FPLC=1569,P<0.01;FHepG2=480.4,P<0.01);Cyclin D1단백하조0.28±0.06、0.15±0.03、0.44±0.04、0.13±0.02,역정제량의뢰관계,여대조조상비차이유통계학의의(P<0.01),량간예조상비차이유통계학의의(FPLC=35.5,P<0.01;FHepG2=114.7,P<0.01).결론 MIF화Cyclin D1재간세포암중과표체,MIF가능재전록수평조공Cyclin D1적표체,병촉사종류세포통과세포주기검측점,계속증치화분화,도치종류적형성.
Objective To investigate the expression of macrophage migration inhibition factor (MIF) and cell cycle regulating factor Cyclin D1 in hepatocellular carcinoma tissue and the interaction between MIF and Cyclin D1 in hepatocellular carcinoma cell cycle controlling. Methods Using quantitative real-time PCR and Western blotting to detect mRNA and protein expression of MIF and Cyelin DI in HCC tissues and tumor adjacent tissues. Specific small interfering RNA(siRNA) targeting MIF gene was transfccted at doses of 50 nmol/L and 100 nmoL/L into HCC cell lines of PLC and HepG2 with lipofeetamine 2000 methods to knockdown the expression of M1F gene and to investigare the the interaction between M1F and Cyclin D1. Results MIF and Cyclin D1 protein and mRNA were overexpressed in HCC tumor tissues. The relative expression of MIF,Cyclin D1 protein and mRNA were 0.825±0.13,0.843± 0.104 and 7.31±1.85 folds、4.27±1.05 folds, compared with the tumor adjacent tissues (FMIF= 15.5, P<0.01;FCyclin D1=87.5,P <0.01). In MIF siRNA treated PLC and HepG2 cells, MIF mRNA down regulation 71.2%±7.2%, 87.4%±2.9% ,74.3%±8.9% and 88.4%±4.6% respectively (FPLC = 315.5 ,P < 0.01 ; FHepG2= 201.2 P < 0.01). While MIF protein expression were significandy reduced to 0.33±0.03,0.11±0.02, 0.81±0.08 and 0.36±0.02 in a dose-dependent manner (FPLC= 43.9, P <0.01 ;FHepG2 = 133.4 P <0.01). Cyclin D1 mRNA was significantly down-regnlated in MIF siRNA treated PLC and HepG2 cell lines when compared with control group(P <0.01). In 50 nmol/L and 100 nmol/L groups, Cyclin DI mRNA levels were respectively decreased by 68.2%±3% and 78.1%±1.4% in PLC cell, 65.8%±4.7% and 77.3%±2.6% in HepG2 cell (FPLC= 1569, P < 0.01 ; FHepG2= 480.4, P <0.01). Compared with control groups, Cyclin D1 protein levels significantly reduced to 0.28±0.06、0.15±0.03 and 0.44 ±0.04、0.13±0.02 in the PLC and HepG2 after M IF siRNA treatment(FPLC= 35.5, P < 0.01 ; FHepG2 = 114.7, P < 0.01). Conclusions MIF and Cyclin D1 mRNA and protein were overexpressed in HCC tumor tissues and participated in tumor cell cycle regulation. MIF may up-regnlate the expression of Cyclin DI via ERK signalling and precipitate in carcinogenesis of hepatocellular carcinoma.
