中华普通外科杂志
中華普通外科雜誌
중화보통외과잡지
CHINESE JOURNAL OF GENERAL SURGERY
2014年
5期
378-381
,共4页
关蛟%张正筠%周尊强%李浩%王书云%周光文
關蛟%張正筠%週尊彊%李浩%王書雲%週光文
관교%장정균%주존강%리호%왕서운%주광문
癌,肝细胞%基因,p53%沉默信息调节因子2相关酶1
癌,肝細胞%基因,p53%沉默信息調節因子2相關酶1
암,간세포%기인,p53%침묵신식조절인자2상관매1
Carcinoma,hepatocellular%Genes,p53%Sirtuin1
目的 研究沉默信息调节因子2相关酶1(silent mating-type information regulation 2 homologue 1,Sirtuin1或SIRT1)在野生型及突变型p53肝细胞肝癌中差异性调节肿瘤形成的机制.方法 对PLC5(249位点突变型p53),HepG2(野生型p53)细胞株进行shSIRT1慢病毒转染,通过Western blot检测信号转导通路,通过细胞生长和增殖实验,集落形成实验,动物实验等检测HepG2细胞,HepG2-shSIRT1细胞,PLC5细胞和PLC5-shSIRT1细胞肿瘤形成能力.结果 抑制SIRT1表达可降低HepG2(野生型p53)肝癌细胞的增殖,提高PLC5(突变型p53)肝癌细胞的增殖(t=3.595,P＜0.01);敲除HepG2细胞中的SIRT1使得p53活化,激活p53下游蛋白p21的表达,敲除PLC5细胞中SIRT1能使乙酰化p53蛋白表达升高,但其下游蛋白p21不能被激活;在HepG2细胞中抑制SIRT1不能改变AMP依赖的蛋白激酶[adenosine 5'-monophosphate (AMP)-activated protein kinase,AMPK]的功能(磷酸化AMPK蛋白),而在PLC5细胞(突变型p53)中抑制SIRT1可降低AMPK的功能(t=4.268,P＜0.01).结论 在肝细胞肝癌中SIRT1激活后的作用及功能主要受到p53突变状态的影响.
目的 研究沉默信息調節因子2相關酶1(silent mating-type information regulation 2 homologue 1,Sirtuin1或SIRT1)在野生型及突變型p53肝細胞肝癌中差異性調節腫瘤形成的機製.方法 對PLC5(249位點突變型p53),HepG2(野生型p53)細胞株進行shSIRT1慢病毒轉染,通過Western blot檢測信號轉導通路,通過細胞生長和增殖實驗,集落形成實驗,動物實驗等檢測HepG2細胞,HepG2-shSIRT1細胞,PLC5細胞和PLC5-shSIRT1細胞腫瘤形成能力.結果 抑製SIRT1錶達可降低HepG2(野生型p53)肝癌細胞的增殖,提高PLC5(突變型p53)肝癌細胞的增殖(t=3.595,P＜0.01);敲除HepG2細胞中的SIRT1使得p53活化,激活p53下遊蛋白p21的錶達,敲除PLC5細胞中SIRT1能使乙酰化p53蛋白錶達升高,但其下遊蛋白p21不能被激活;在HepG2細胞中抑製SIRT1不能改變AMP依賴的蛋白激酶[adenosine 5'-monophosphate (AMP)-activated protein kinase,AMPK]的功能(燐痠化AMPK蛋白),而在PLC5細胞(突變型p53)中抑製SIRT1可降低AMPK的功能(t=4.268,P＜0.01).結論 在肝細胞肝癌中SIRT1激活後的作用及功能主要受到p53突變狀態的影響.
목적 연구침묵신식조절인자2상관매1(silent mating-type information regulation 2 homologue 1,Sirtuin1혹SIRT1)재야생형급돌변형p53간세포간암중차이성조절종류형성적궤제.방법 대PLC5(249위점돌변형p53),HepG2(야생형p53)세포주진행shSIRT1만병독전염,통과Western blot검측신호전도통로,통과세포생장화증식실험,집락형성실험,동물실험등검측HepG2세포,HepG2-shSIRT1세포,PLC5세포화PLC5-shSIRT1세포종류형성능력.결과 억제SIRT1표체가강저HepG2(야생형p53)간암세포적증식,제고PLC5(돌변형p53)간암세포적증식(t=3.595,P＜0.01);고제HepG2세포중적SIRT1사득p53활화,격활p53하유단백p21적표체,고제PLC5세포중SIRT1능사을선화p53단백표체승고,단기하유단백p21불능피격활;재HepG2세포중억제SIRT1불능개변AMP의뢰적단백격매[adenosine 5'-monophosphate (AMP)-activated protein kinase,AMPK]적공능(린산화AMPK단백),이재PLC5세포(돌변형p53)중억제SIRT1가강저AMPK적공능(t=4.268,P＜0.01).결론 재간세포간암중SIRT1격활후적작용급공능주요수도p53돌변상태적영향.
Objective To understand how SIRT1 differently regulates oncogenesis in hepatocellular carcinoma (HCC) with wild type and mutant type p53.Methods HCC cell line PLC5 cells (249 site mutated p53),and HepG2 cells (wild type p53) were infected with lentivirus containing shSIRT1.Western blotting was used for signaling pathway detection.Cell growth and proliferation assay,colony formation assay and tumor xenograft assay were performed to test the tumor growth ability of HepG2 cells,HepG2-shSIRT1 cells,PLC5 cells and PLC5-shSIRT1 cells respectively.Results SIRT1 silencing resulted in significant inhibition of cell proliferation in HepG2 cells but stimulating cell proliferation in PLC5 cells (t =3.595,P ＜0.01).Acetylation of p53 was found in HepG2 (HepG2-shSIRT1) and p21 was up-regulated,however,in PLC5 (PLC5-shSIRT1) cells,acetylation of p53 was found but p21 was not induced despite of p53 activation.Silence of SIRT1 resulted in no change of AMPK function in HepG2 cells but a lower activity of AMPK in PLC5 cells (t =4.268,P ＜ 0.01).Conclusions In HCC cell lines the function following SIRT1 activation is largely determined by p53 mutant status.
