简介:背景:Gankyrin是一个含锚蛋白重复序列的原癌蛋白,其高表达参与了多种恶性肿瘤的发生、发展进程。目的:探讨下调gankyrin表达对胃癌细胞增殖能力的影响及其可能机制。方法:以携带gankyrinsiRNA的慢病毒载体转染人胃癌细胞株MKN28,分别采用MTT实验、流式细胞术和蛋白质印迹法检测下调gankyrin表达对MKN28细胞增殖、细胞周期分布及其β-catenin/cyclinD1信号通路的影响。结果:慢病毒载体的转染效率在90%以上,转染gankyrinsiRNA后,MKN28细胞gankyrin蛋白表达显著受抑(P<0.01)。与未转染慢病毒和转染对照病毒的细胞相比,转染gankyrinsiRNA的MKN28细胞体外生长于第3天起显著受抑,细胞周期G1期细胞比率增高,S期细胞比率降低,细胞中的β-catenin和cyclinD1表达水平降低,差异均有统计学意义(P<0.01)。结论:下调胃癌细胞中的gankyrin表达可通过抑制β-catenin/cyclinD1信号通路引起细胞周期G1期阻滞和细胞增殖抑制,gankyrin有望成为胃癌靶向治疗的新靶点。
简介:目的观察早期胃癌组织中GTP酶激活蛋白SH3功能区结合蛋白G3BP1和G3BP2的表达变化并探讨其意义。方法选择早期胃癌组织89例和正常对照组织35例,采用免疫组织化学方法检测两组G3BP1蛋白和G3BP2蛋白的表达情况。结果早期胃癌组和正常组G3BP1的阳性率分别为87.64%和60.00%,G3BP2阳性率分别为86.51%和54.28%,两组比较差异有统计学意义(P=0.000,0.000);早期胃癌G3BP1和G3BP2的表达均与幽门螺杆菌感染相关(P=0.000);早期胃癌组织中G3BP1与G3BP2表达呈正相关(rs=0.252,P=0.017)。结论早期胃癌组织中G3BP1和G3BP2呈高表达,可能与幽门螺杆菌感染紧密相关,两指标异常表达可能在早期胃癌的发生和发展中起重要作用。
简介:目的观察MiR-384对HFFA诱导的Hepa1-6细胞sirt3/FOXO1信号通路的影响。方法取Hepa1-6细胞,加入OA和PA储存液,使两者终浓度分别为1mmol/L和0.5mmol/L,培养24h,建立非酒精性脂肪性肝炎体外细胞模型。测定肝细胞内活性氧水平评估模型建立的情况。分别以miR-384模拟物、miR-ctrl、miR-384抑制剂、miR-384抑制剂-ctrl、沉默信息调节因子3(Sirt3)或si-ctrl转染细胞48h。使用FCM测定各组细胞ROS水平,采用Westernblot法检测各组细胞sirt3、FOXO1及抗氧化蛋白锰超氧化物歧化酶(MnSOD)和抗氧化蛋白过氧化氢酶(CAT)表达,采用专用试剂盒检测SOD和CAT活性。结果与正常组(0.66±0.01)比,miR-384模拟物组和sisirt3组细胞内活性氧(ROS)水平显著升高[分别为(37.3±1.13)和(10.4±0.36),P<0.01];与HFFA组(29.4±0.98)比,HFFA/miR-384抑制剂组细胞ROS水平显著降低[(12.8±0.41),P<0.01];与正常组(0.75±0.04)Hepa1-6细胞sirt3蛋白表达水平比,HFFA组显著降低[(0.23±0.01),P<0.01];与HFFA组比,HFFA/sirt3组显著增加[(0.83±0.03),P<0.01];与HFFA/sirt3组比,HFFA/sirt3/miR-384组显著降低[(0.46±0.02),P<0.01];与对照组比,miR-384模拟物组Hepa1-6细胞sirt3蛋白、Forkhead转录因子O亚家族(FOXO)成员FOXO1蛋白表达显著减少[分别为(0.2±0.01)和(0.3±0.01),P<0.01],而CAT和MnSOD表达显著增加[分别为(2.3±0.05)和(2.4±0.06),P<0.01];与HFFA组比,HFFA/miR-384抑制剂组Hepa1-6细胞sirt3蛋白和FOXO1蛋白表达显著增加[分别为(0.5±0.02)和(0.7±0.01),P<0.01],MnSOD和CAT表达水平显著降低[分别为(1.6±0.04)和(2.0±0.03),P<0.01];与NAFLD组SOD(327±3.45)和CAT(386±4.03)活性比,正常组SOD和CAT[分别为(425±5.49)和(512±6.04),P<0.01]和miR-384抑制剂组SOD和CAT活性[分别为(406±4.79)和(447±5.38),P<0.01]显著升高。结论miR-384表达可导致HFFA诱导的Hepa1-6细胞氧化损伤加重,部分可能是通过抑制sirt3/FOXO1途径实现的。
简介:背景:聚腺苷二磷酸核糖聚合酶-1(PARP-1)主要参与对DNA损伤的反应,caspase-3主要参与细胞凋亡的调控,两者与结直肠癌发生的关系尚不十分清楚。目的:探讨PARP-1和caspase-3在结直肠腺瘤和腺癌中的表达及其与肿瘤临床病理特征的关系。方法:以免疫组化方法检测59例结直肠腺瘤、70例结直肠腺癌和10例癌旁黏膜石蜡包埋标本中PARP-1和caspase-3的表达,分析腺瘤和腺癌组织中两者表达与肿瘤临床病理参数的相关性,以及两者间的相关性。结果:结直肠腺瘤和腺癌组织中,PARP-1的阳性表达率显著高于癌旁结直肠黏膜(55.9%和82.9%对20.0%,P〈0.05),caspase-3的阳性表达率显著低于癌旁结直肠黏膜(33.9%和28.6%对100.0%,P〈0.05)。结直肠腺瘤中,PARP-1的表达与腺瘤数目相关,caspase-3的表达与上皮内瘤变程度相关;结直肠腺癌中,PARP-1的表达与癌组织分化程度相关。PARP-1与caspase-3在结直肠腺瘤和腺癌组织中的表达无相关性。结论:PARP-1和caspase-3在结直肠腺癌的发生、发展过程中起有重要作用,两者联合检测对结直肠腺癌的早期诊断具有一定价值。
简介:180例幽门螺杆菌(Hp)感染的消化性溃疡或非溃疡性消化不良(NUD)患者分3组接受胶体次拘椽酸铋(CBS)合并的两联治疗。A组方案为CBS240mg,交沙霉素1000mg;B组为CBS240mg,强力霉素100mg;C组(对照组)为CBS240mg,羟氨苄青霉索l000mg。上述剂量每日2次,疗程2周。162例完成治疗后停药4周后胃镜复查。结果显示:A、B和C组的Hp根除率分别为69.6%(39/56)、16.3%(9/55)和43.1%(22/51),各组间的Hp根除率有非常显著差别(P<0.01或P<0.001):A、B和C组的十二指肠溃疡(DU)愈合率分别为86.9%(20/23)、60.8%(14/23)和75.O%(12/16),A组的DU愈合率显著高于B组(P
简介:AIM:Toevaluatetheefficacyofthe14-dmoxifloxacinbasedtripletherapyforthesecond-lineeradicationofHelicobacterpylori(H.pylori)infection.METHODS:Between2011and2013,weconductedaretrospectivereviewofthemedicalrecordsof160patientswhohadexperiencedfailureoftheirfirst-lineprotonpumpinhibitor-basederadicationtherapyandsubsequentlyreceivedthemoxifloxacin-basedtripletherapyasasecond-lineeradicationtreatmentregimen.Thepatientswhoweretreatedwiththemoxifloxacinbasedtripletherapy(oral20mgrabeprazoleb.i.d.,1000mgamoxicillinb.i.d.,and400mgmoxifloxacinq.d.)for7dwereassignedtotheRAM-7group(n=79)whilethosewhotookthemfor14dayswereassignedtoRAM-14group(n=81).Theeradicationratesforbothgroupsweredeterminedbyintentionto-treat(ITT)andper-protocol(PP)analyses.ITTanalysiscomparedthetreatmentgroupsasoriginallyallocatedwhilethePPanalysisincludingonlythosepatientswhohadcompletedthetreatmentasoriginallyallocated.SuccessfuleradicationtherapyforH.pyloriinfectionwasdefinedasthedocumentationofanegative13C-ureabreathtest4wkaftertheendoftheeradicationtreatment.RESULTS:TheoverallITTeradicationratewas76.2%(122/160).ThefinalITTeradicationrateswere70.8%(56/79;95%CI:63.3%-77.1%)intheRAM-7groupand81.4%(66/81;95%CI:74.6%-88.3%)intheRAM-14group(P=0.034).TheoverallPPeradicationratewas84.1%(122/145),andthefinalPPeradicationrateswere77.7%(56/72;95%CI:70.2%-85.3%)intheRAM-7groupand90.4%(66/73;95%CI:82.8%-98.1%)intheRAM-14group(P=0.017).TheH.pylori-eradicationratesintheRAM-14groupweresignificantlyhighercomparedwiththatoftheRAM-7groupaccordingtoboththeITT(P=0.034)andthePPanalyses(P=0.017).Bothgroupsexhibitedgoodtreatmentcompliance(RAM-7/RAM-14group:100%/100%).Theadverseeventrateswere19.4%(14/72)and20.5%(15/73)intheRAM-7andRAM-14groups,respectively(P=0.441).Adverseeventsoccurredin14
简介:AIM:Toinvestigatetheefficacyofneoadjuvantchemoradiotherapy(NACRT)forresectabilityoflocallyadvancedgastriccancer(LAGC).METHODS:BetweenNovember2007andJanuary2014,29patientswithLAGC(clinicallyT3withdistalesophagusinvasion/T4orbulkyregionalnodemetastasis)thatweretreatedwithNACRTfollowedbyD2gastrectomywereincludedinthisstudy.ResectabilitywasevaluatedwithradiologicandendoscopicexamsbeforeandafterNACRT.Usingthreedimensionalconformalradiotherapy,patientsreceived45Gy,withadailydoseof1.8Gy.Theentiretumorextentandtheregionalmetastaticlymphnodeswereincludedinthegrosstumorvolume.PatientspresentingwitharesectabletumorafterNACRTreceivedatotalorsubtotalgastrectomywithD2dissection.ThepathologictumorresponsewasevaluatedusingJapaneseGastricCancerAssociationhistologicevaluationcriteria.PostoperativemorbiditywasevaluatedusingtheNationalCancerInstitute-CommonTerminologyCriteriaforAdverseEventsversion4.0.Overallsurvival(OS)andprogression-freesurvival(PFS)rateswereestimatedusingaKaplan-Meieranalysisandcomparedusingthelog-ranktest.RESULTS:Allpatientswereassessedasunresectablecases.Twenty-fourpatients(24/29;82.8%)showedLAGConpositronemissiontomography-computedtomography(CT)andcontrast-enhancedCT,whereasfourpatients(4/29;13.8%)withvagueinvasionorabutmenttoanadjacentorganunderwentdiagnosticlaparoscopy.Onepatient(1/29;3.4%),initiallyassessedasaresectablecase,underwentan'openandclosure'afterthetumorwasfoundtobeunresectable.Abutmenttoanadjacentorgan(34.5%)wasthemostcommonreasonforNACRT.TheclinicalresponserateonemonthafterNACRTwas44.8%.AfterNACRT,69%(20/29)ofpatientshadaresectabletumor.Ofthe20patientswitharesectabletumor,18patients(62.1%)underwentaD2gastrectomy.TheR0resectionratewas94.4%andtwopatients(2/18;11.1%)showedacompleteresponse.Themedianfollow-updurationwas13.5mo.Theone-yearOSandPFS
简介:AIM:ToinvestigatetheroleoftheoverexpressionofB7-H3inapoptosisincolorectalcancercelllinesandtheunderlyingmolecularmechanisms.METHODS:SW620cellsthathighlyoverexpressedB7-H3(SW620-B7-H3-EGFP)andHCT8cellsstablytransfectedwithB7-H3shRNA(HCT8-shB7-H3)werepreviouslyconstructedinourlaboratory.CellstransfectedwithpIRES2-EGFPwereusedasnegativecontrols(SW620-NCandHCT8-NC).Real-timePCRandwesternblottinganalysiswereusedtodetectthemRNAandproteinexpressionsoftheapoptosisregulatorproteinsBcl-2,Bcl-xlandBax.Acellproliferationassaywasusedtoevaluatethesurvivalrateanddrugsensitivityofthecells.Theeffectofdrugresistancewasdetectedbyacellcycleassay.Activecaspase-3westernblottingwasusedtoreflecttheanti-apoptoticabilityofcells.WesternblottingwasalsoperformedtodeterminetheexpressionofproteinsassociatedwiththeJak2-STAT3signalingpathwayandtheapoptosisregulatorproteinsafterthetreatmentwithAG490,aJak2specificinhibitor,inB7-H3overexpressingcells.ThedatawereanalyzedbyGraphPadPrism6usinganon-pairedt-test.RESULTS:WhetherbyoverexpressioninSW620cellsordownregulationinHCT8,B7-H3significantlyaffectedtheexpressionofanti-andpro-apoptoticproteins,atboththetranscriptionalandtranslationallevels,comparedwiththenegativecontrol(P<0.05).AcellproliferationassayrevealedthatB7-H3overexpressionincreasedthedrugresistanceofcellsandresultedinahighersurvivalrate(P<0.05).Inaddition,theresultsofcellcycleandactivecaspase-3westernblottingprovedthatB7-H3overexpressioninhibitedapoptosisincolorectalcancercelllines(P<0.05).B7-H3overexpressionimprovedJak2andSTAT3phosphorylationand,inturn,increasedtheexpressionofthedownstreamanti-apoptoticproteinsB-cellCLL/lymphoma2(Bcl-2)andBcl-xl,basedonwesternblotting(P<0.05).AftertreatingB7-H3overexpressingcellswiththeJak2-specificinhibitorAG490,thephosphorylationofJak2andSTAT3,andtheexp
简介:瞄准:与ulcerative(UC)在病人的结肠的粘膜检验metalloproteinase-1(MMP-1)和metalloproteinase-1(TIMP-1)的织物禁止者的表示。方法:颠倒抄写聚合酶链反应(RT-PCR),免疫组织化学被用来与UC和控制在病人在mRNA和蛋白质层次学习MMP-1和TIMP-1的表达式。在MMP-1mRNA,TIMP-1mRNA,MMP-1mRNA/TIMP-1mRNA比率和有UC的病人的临床的症状的严厉之间的关系也被分析。结果:MMP-1mRNA和TIMP-1mRNA在的表示溃烂并且煽动了结肠的粘膜在非煽动的结肠的粘膜比那显著地高(P<0.001),但是有不,统计上,在UC病人和正常的非煽动的结肠的粘膜的有效差量控制(P>0.05)。在UC病人的溃烂的结肠的粘膜的MMP-1和TIMP-1的mRNA表达式被80褶层和2.2褶层增加,分别地什么时候与正常控制相比。在煽动的结肠的粘膜,增加分别地是30褶层和1.6褶层。Immunohistochemical分析显示出那在之中溃烂,发炎,并且UC病人的非煽动的结肠的mucosae和正常控制,MMP-1表示的积极的率分别地是87%,87%,40%和35%,并且TIMP-1表示的积极的率分别地是89%,89%,80%和75%。而且,MMP-1mRNA,TIMP-1mRNA和MMP-1mRNA/TIMP-1mRNA比率的表示与临床的症状(P<0.05)的严厉被相关。结论:在结肠的粘膜在UC病人引起细胞外的矩阵(ECM)和溃疡的过多的水解作用的diseased的MMP-1的过多的表示。MMP-1mRNA,TIMP-1mRNA和MMP-1mRNA/TIMP-1mRNA比率能被用作简历标记与UC在病人判定临床的症状的严厉。外长的TIMP-1或MMP-1禁止者治疗是为有UC的病人的新奇治疗。
简介:AIMToinvestigatetherolesandinteractionsofmutThomolog(MTH)-1andhypoxia-induciblefactor(HIF)-1αinhumancolorectalcancer(CRC).METHODS:TheexpressionanddistributionofHIF-1αandMTH-1proteinsweredetectedinhumanCRCtissuesbyimmunohistochemistryandquantitativerealtimepolymerasechainreaction(qRT-PCR).SW480andHT-29cellswereexposedtonormoxiaorhypoxia.ProteinandmRNAlevelsofHIF-1αandMTH-1wereanalyzedbywesternblottingandqRT-PCR,respectively.InordertodeterminetheeffectofHIF-1αontheexpressionofMTH-1andtheamountof8-oxodeoxyguanosinetriphosphate(dGTP)inSW480andHT-29cells,HIF-1αwassilencedwithsmallinterferingRNA(siRNA).GrowthstudieswereconductedoncellswithHIF-1αinhibitionusingaxenografttumormodel.Finally,MTH-1proteinwasdetectedbywesternblottinginvivo.RESULTS:HighMTH-1mRNAexpressionwasdetectedin64.2%ofcases(54/84),andthiswassignificantlycorrelatedwithtumorstage(P=0.023)andsize(P=0.043).HIF-1αproteinexpressionwascorrelatedsignificantlywithMTH-1expression(R=0.640;P〈0.01)inhumanCRCtissues.HypoxicstressinducedmRNAandproteinexpressionofMTH-1inSW480andHT-29cells.InhibitionofHIF-1αbysiRNAdecreasedtheexpressionofMTH-1andledtotheaccumulationof8-oxo-dGTPinSW480andHT-29cells.Intheinvivoxenografttumormodel,expressionofMTH-1wasdecreasedintheHIF-1αsiRNAgroup,andthetumorvolumewasmuchsmallerthanthatinthemocksiRNAgroup.CONCLUSION:MTH-1expressioninCRCcellswasupregulatedviaHIF-1αinresponsetohypoxicstress,emphasizingthecrucialroleofHIF-1α-inducedMTH-1intumorgrowth.