timp-3对肝癌细胞增殖、凋亡及侵袭转移的影响

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1、广东医学2014年1月第35卷第1期GuangdongMedicalJournalJan．2014，Vo1．35，No．I}==影·29·响一3对肝癌细胞增殖、凋亡及侵袭转移的：l：陈沅然，沈波，李瑜元广州医科大学附属广州市第一人民医院消化科、广州市临床医学研究所(510180)【摘要】目的建立以真核表达载体介导的过表达TIMP一3基因的肝癌细胞株(SMMC一7721和HepG2)，探讨TIMP一3基因对两细胞增殖、凋亡、周期、侵袭等功能的影响。方法将TIMP一3质粒pCMV6一AC—GFP／TIMP一3(TIMP一3组)和空载对照质粒pCMV6一AC—GFP(对照组

2、)稳定转染入SMMC一7721和HepG2细胞株，用Westernblot技术检测细胞内TIMP一3蛋白的表达，通过MTS实验、流式细胞术、侵袭实验测定细胞的各种生物学行为。结果Westernblot检测结果显示，TIMP一3组的SMMC一7721和HepG2细胞高表达TIMP一3蛋白，而对照组细胞没有TIMP一3蛋白表达。MTS实验和流式细胞术检测结果显示，与对照组比较，TIMP一3组两种细胞的增殖能力显著减弱(第5天时P=0．003，P=0．009)，凋亡率显著升高(P=0．001，P=0．038)，细胞周期停滞于G2／M期(P=0．030，P=0．034)；体外

3、侵袭实验结果显示，TIMP一3组两种细胞的侵袭能力较对照组显著下降(P=0．033，P=0．015)。结论TIMP一3能显著抑制SMMC一7721及HepG2细胞的增殖和侵袭能力，并诱导细胞发生凋亡和产生G，／M期阻滞．【关键词】TIMP一3；肝癌；增殖；凋亡；细胞周期；侵袭TheeffectsofTIMP一3ontheproliferation，apoptosis，invasivenessandmigrationofhumanhepatocarcinomacellline．CHENYuan—ran，SHENBo，L1一yuan．DepartmentofGastroe

4、nterology，GuangzhouFirstPeoplesHospi—tal，GuangzhouMedicalUniversity，GuangzhouInstituteofClinicalMedicine，Guangzhou510180，ChinaCorrespondingauthor：LIYu—yuan．E—mail：liyyliyy@tom．corn【Abstract】ObjectiveToconstructastableTIMP一3over—expressionhepatocellularcarcinoma(HCC)celllines(SMMC一7721an

5、dHepG2)forinvestigatingtheefectsofTIMP一3onproliferation，apoptosis，cellcycleandinva—sivenessofthetwocelllines．MethodsThepCMV6一AC—GFP／TIMP一3andemptyplasmidpCMV6一AC—GFP(contro1)werebothtransfectedintoSMMC一7721andHepG2cells．TheTIMP一3proteinwasassessedbyWesternblot．TheeffectsofTIMP一3onthebio

6、logicalbehaviorsofSMMC一7721andHepG2weretestedbyMTSassay，floweytome—try．andinvasivenessassay．ResultsOver—expressionofTIMP一3wasobservedinSMMC一7721andHepG2cellstransfeetedwithTIMP一3，whilenoexpressionofTIMP一3wasrevealedincontrols．TheproliferationandinvasivenessofSMMC一7721andHepG2cellsweresi

7、gnificantlysuppressedbyTIMP一3over—expression．bywhichsignificantlyin—creasedapoptosisrateandG2／Mcellcyclearrestwerealsoobserved(P<0．05)．ConclusionTIMP一3significantlyin—hibitsthecapabilitiesofproliferationandinvasivenessofSMMC一7721andHepG2cells，inducesHCCcellsapoptosisandcausedce