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时间:2020-05-04
《PTEN/Akt通路抑制人肝癌HepG2细胞增殖-论文.pdf》由会员上传分享,免费在线阅读,更多相关内容在应用文档-天天文库。
1、细胞与分子免疫学杂志(ChinJCellMolImmuno1)2014.30(2147·论著·文章编号:1007—8738(2014)02—0147—04罗格列酮通过PI3PTEN/Akt通路抑制人肝癌HepG2细胞增殖张萌,彭利,乔治斌,何宏涛,周烨,徐卓(河北医科大学第四医院,河北石家庄050011)[摘要]目的研究罗格列酮对人肝癌HepG2细胞增殖与细胞周期的影响,通过检测相关蛋白的表达变化,探讨其可能机制。方法不同浓度罗格列酮作用于HepG2细胞,M'VF法测定细胞增殖抑制率,流式细胞术检测细胞周期的变化,Westernblot法检测PTEN、pAkt
2、、S期激酶相关蛋白2(skp2)及P'27蛋白表达变化,RT.PCR检测PTEN、Skp2及P27p1mRNA表达变化。结果罗格列酮可明显抑制HepG2细胞的增殖,呈现时间和浓度依赖性(P<0.05);GO/G1期HepG2细胞比例明显升高,s期细胞比例明显降低(P<0.05)。Westernblot结果显示罗格列酮可下调HepG2细胞中pAkt和skp2蛋白的表达,上调PTEN和P27蛋白表达(P<0.05)。RT—PCR结果显示罗格列酮可明显上调PTENmRNA的表达,下调skp2mRNA的表达,而P27“mRNA表达无明显变化。结论罗格列酮可抑制HepG
3、2细胞的增殖,造成G0/G1期阻滞,其机制可能与罗格列酮经PI3K/trI'EN/Akt信号通路参与skp2及P27蛋白的调控有关。[关键词]罗格列酮;PTEN;PI3K/Akt信号通路;肝细胞癌;凋亡;S期激酶相关蛋白2;P27“[中图分类号]R735.7,Q255,R965[文献标志码]ARosiglitazoneinhibitshumanHepG2cellproliferationv/aPDK/PTEN/AktsignalingpathwayZHANGMeng,PENGLi,QIAOZhibin,HEHongtao,ZHOUYe,XUZhuoDepart
4、mentofHepatobiliarySurgery,FourthAfiliatedHospital,HebeiMedicalUniversity,Shijiazhuang050011,China[Abstract]0bjeetiveToinvestigatetheefectsofrosiglitazone(ROlz)onproliferationandcellcycleofhumanhepatocallularcarcinomacalllineHepG2andexploretheunderlyingmechanismsbydetectingtherelate
5、dproteins.MethodsAftertreatedwithROZofdiferentconcentrations,HepG2cellsweretestedforthechangesinthecellproliferationbyM3-1-assayandinthecelcyclebyflowcytometry.WesternblottingandRT—PCRwereperformedtomeasuretheexpressionsofPTEN,pARt,Sphasekinaseassociatedprotein2(Skp2)andP27atprotein
6、andmRNAlevels,respectively.ResultsROZsignificantlyinhibitedHepG2celIproliferationinaconcentration·dependentandtime-dependentmanner(P<0.05)。TheproportionofHepG2cellsinG0/G1phaseincreased,andthatinSphasedecreasedsignificantly(P<0.05).ROZreducedtheexpressionsofpAktandSkp2,andraisedthee
7、xpressionsofPTENandP27inHepG2cells(P<0.05).RT—PCRrevealedthatROZjncreasedtheexpressionsofPTENmRNA,decreasedtheexpressionsofSkp2mRNA,andhadnoefe~onP27mRNA.Condusion0urstudydemonstratedthatReZcouldinhib~HepG2celIproliferationandblockG0/G1phase,themechanismsmayberelatedtotheregulationo
8、ntheexpressionsofSk
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