中华消化病与影像杂志（电子版）
中華消化病與影像雜誌（電子版）
중화소화병여영상잡지（전자판）
2011年
1期
7-9
,共3页
曲建慧%楼敏%张敏娜%许桂林%常秀娟%陈艳%周霖%高旭东%杨永平
麯建慧%樓敏%張敏娜%許桂林%常秀娟%陳豔%週霖%高旭東%楊永平
곡건혜%루민%장민나%허계림%상수연%진염%주림%고욱동%양영평
肝癌细胞系,干扰素-α%HepG2%抑癌基因%DKK-1
肝癌細胞繫,榦擾素-α%HepG2%抑癌基因%DKK-1
간암세포계,간우소-α%HepG2%억암기인%DKK-1
Hepatocellular carcinoma cell line,Interferon-α%HepG2%Tumor suppressor gene%Dickkopf-1
目的 探讨重组干扰素-α (IFN-α)对肿瘤抑制作用的靶分子及可能作用机制.方法 选用HepG2肝癌细胞系,分别应用1000 U/ml、2000 U/ml浓度的IFN -α作用于HepG2细胞,在药物作用6h、16 h后应用RT-PCR；Dickkopf-1(DKK-1)方法分别检测各实验组DKK-1 mRNA表达水平,Western blot杂交法检测DKK-1蛋白表达水平.结果 定量RT-PCR结果显示,IFN-α处理能增强DKK-1mRNA表达.2000 U/mL IFN-α作用HepG2细胞16h组其DKK-1 mRNA表达(0.00039±0.000057)高于处理6h组(0.000195±0.000021,t=6.994,P=0.0022)以及1000 U/mL作用16 h组(0.000307±0.000012,t=2.876,P=0.0452); Western blot显示重组IFN-α作用于HepG2细胞后,均能够导致细胞内DKK-1蛋白表达增强(P值均＜0.05).结论 IFN -α能够上调DKK-1表达,DKK-1可能是IFN -α作用的下游调节靶基因,这可能是IFN-α发挥抗肿瘤作用机制之一.
目的 探討重組榦擾素-α (IFN-α)對腫瘤抑製作用的靶分子及可能作用機製.方法 選用HepG2肝癌細胞繫,分彆應用1000 U/ml、2000 U/ml濃度的IFN -α作用于HepG2細胞,在藥物作用6h、16 h後應用RT-PCR；Dickkopf-1(DKK-1)方法分彆檢測各實驗組DKK-1 mRNA錶達水平,Western blot雜交法檢測DKK-1蛋白錶達水平.結果 定量RT-PCR結果顯示,IFN-α處理能增彊DKK-1mRNA錶達.2000 U/mL IFN-α作用HepG2細胞16h組其DKK-1 mRNA錶達(0.00039±0.000057)高于處理6h組(0.000195±0.000021,t=6.994,P=0.0022)以及1000 U/mL作用16 h組(0.000307±0.000012,t=2.876,P=0.0452); Western blot顯示重組IFN-α作用于HepG2細胞後,均能夠導緻細胞內DKK-1蛋白錶達增彊(P值均＜0.05).結論 IFN -α能夠上調DKK-1錶達,DKK-1可能是IFN -α作用的下遊調節靶基因,這可能是IFN-α髮揮抗腫瘤作用機製之一.
목적 탐토중조간우소-α (IFN-α)대종류억제작용적파분자급가능작용궤제.방법 선용HepG2간암세포계,분별응용1000 U/ml、2000 U/ml농도적IFN -α작용우HepG2세포,재약물작용6h、16 h후응용RT-PCR；Dickkopf-1(DKK-1)방법분별검측각실험조DKK-1 mRNA표체수평,Western blot잡교법검측DKK-1단백표체수평.결과 정량RT-PCR결과현시,IFN-α처리능증강DKK-1mRNA표체.2000 U/mL IFN-α작용HepG2세포16h조기DKK-1 mRNA표체(0.00039±0.000057)고우처리6h조(0.000195±0.000021,t=6.994,P=0.0022)이급1000 U/mL작용16 h조(0.000307±0.000012,t=2.876,P=0.0452); Western blot현시중조IFN-α작용우HepG2세포후,균능구도치세포내DKK-1단백표체증강(P치균＜0.05).결론 IFN -α능구상조DKK-1표체,DKK-1가능시IFN -α작용적하유조절파기인,저가능시IFN-α발휘항종류작용궤제지일.
Objective To further confirm the upregulation expression of tumor suppressor, Dickkopf 1 (DKK-1 ), in IFN-α treated HepG2 cells. Methods HepG2 cells were treated by various concentration of IFN- α of 1000U/ml, 2000U/ml, respectively. At the time points of 6h、16h after IFN-α treatment, reversetranscription polymerase chain reaction (RT-PCR) and quantitative real-time PCR were performed to detect the expression of DKK-1 Mran, western blot was used to observe the DKK -1 protein levels. Results Realtime PCR showed that HepG2 cells treated with 2000 U/ml IFN-α for 16 h had higher DKK-1 Mran expression 0.00039 ± 0.000057 than cells in control group treated with 2000 U/ml for 6 h 0.000195 ± 0.000021 or 1000 U/ml IFN-α for 16 h 0.000307 ± 0.000012 P＜0.05;Westem blot further confirmed that IFN-treatment increased the expression of DKK-1 protein P＜0.05.Conclusion IFN-α could induce the expression of DKK-1 in HepG2 cells. DKK-1 may be a potential molecular target for IFN- α exerting its anti-proliferative function.
