简介:Objective:Toinvestigatethepost-transcriptionalregulationofp21WAF1/CIP1byp53.Methods:TheMDA-MB-468cellshaveendogenousmutantp53andtheMCF7cellslineshavewtp53.Recombinantp53expressionandp21WAF1/CIP1inductionweredetectedbyWesternblotanalysis.Northernblotanalysiswascarriedouttoexaminewhetherchangesinp21WAF1/CIP1proteinlevelsinMCF7cellstreatedwithAdCMVp53arereflectedatthemRNAlevel.FlowcytometricanalysisofMCF7cellsfollowingoverexpressionofrecombination.Results:Theratioofp53:p21WAF1/CIP1wasbelow1attheearlystagesofAdCMVp53infection,butincreasedto1.6byday3andto9.7byday5post-infection.Asexpected,p21WAF1/CIP1expressionwasnotdetectableinMDA-MB-468cellsdespitethepresenceofhighlevelsofmutantp53protein.TheG1/SratiosinuntreatedcontrolsandAdCMVβgalinfectedMCF7cellswere1.10and1.35,respectively.ByNorthernblotanalyzingthep21WAF1/CIP1:GAPDHratiosatdifferenttimepointsagainsttheratioattimepoint0,amaximum3-foldinductionofp21WAF1/CIP1mRNAexpressionrelativetountreatedcontrolwasobservedonday1post-infection.TheflowcytometricanalysisindicatedthatMCF7cellsinfectedwithAdCMVp53undergoG1arrestatbothtimepointsstudied,withG1/Sratiosrangingfrom5.54atday1to5.65atday7.TheG1/SratiosinuntreatedcontrolsandAdCMVβgalinfectedMCF7cellswere1.10and1.35,respectively.Conclusion:Thisstudydemonstratedthatp53couldregulatep21WAF1/CIP1geneexpressionatboththetranscriptionalandpost-transcriptionallevelsinMCF7cells.Thelattermechanismmaybeinvolvedinorberesponsiblefor,theinductionofcellcyclearrestbytranscription-defectivemutantsofp53.
简介:目的腹腔镜根治性远端胃切除(radicallaparoscopicdistalgastrectomy,LG)+D2淋巴结清扫术用于治疗进展期胃癌其安全性和有效性仍存在争议。本研究设计了一个前瞻性随机对照研究来对比腹腔镜手术和传统开腹手术行根治性远端胃切除术+D2淋巴结清扫术在治疗进展期胃癌的安全性和疗效。研究对象和方法自2012年9月至2014年12月,1056例临床分期为T(2-4a)N0-3M0的胃癌患者被纳入本研究。患者被随机分为腹腔镜根治性远端胃切除术+D2淋巴清扫组(LG组)(n=528)和开腹远端胃切除术(opengastrectomy,OG)+D2淋巴清扫组(OG组)(n=528)。共有15位来自中国14个研究中心的经验丰富的外科医生参与此次研究。根据意向性治疗分析原则比较LG组(n=519)和OG组(n=520)手术后30天内的并发症发生率和死亡率。术后并发症严重程度分级根据Clavien-Dindo分级方法。结果LG组和OG组患者的D2淋巴结清扫术的合格率相似(99.4%vs.99.6%;P=0.845)。术后并发症发生率在LG组为15.2%,OG组为12.9%,统计学无明显差异(相差2.3%,95%CI:-1.9-6.6,P=0.285)。死亡率在LG组为0.4%,而OG组无一例死亡(相差0.4%,95%CI:-0.4-1.4,P=0.249)。两组并发症的严重程度分布相似(P=0.314)。结论有经验的外科医生可以在进展期胃癌患者上安全开展腹腔镜下根治性远端胃切除术+D2淋巴结清扫。
简介:目的探讨粪便DNA中联合分泌型卷曲相关蛋白1(SFRP1)及胞裂蛋白9(SEPT9)基因甲基化检测对老年性结直肠癌(CRC)早期诊断的临床价值。方法选取我院从2016年1月至2016年9月确诊的35例年龄〉60岁CRC患者为研究组,选取同期35例〉60岁正常体检者为对照组。提取CRC患者和正常体检者粪便DNA,采用甲基化特异性PCR(MSP)技术分别检测SFRP1及SEPT9基因的甲基化状态;比较CRC患者单基因和联合双基因甲基化检出率的差异,以及CRC患者联合双基因甲基化检出率与对照组的差异;分析CRC患者SFRP1或SEPT9基因甲基化与临床病理特征(如年龄、性别、肿瘤位置)的关系。结果CRC患者粪便DNA中SFRP1及SEPT9基因甲基化检出率分别为45.71%(16/35)和51.43%(18/35),均显著高于对照组的8.57%(3/35)和14.29%(5/35),差异均有统计学意义(P〈0.01);联合双基因检测结果显示,82.86%(29/35)的CRC患者粪便DNA中至少存在一个基因的甲基化,显著高于单基因检出率(P〈0.01),亦高于对照组的17.14%(6/35),差异有统计学意义(P〈0.01)。CRC患者SFRP1或SEPT9基因甲基化与年龄、性别、肿瘤位置均无关(P〉0.05)。结论粪便DNA中联合SFRP1及SEPT9基因甲基化检测比单基因检测更加敏感,对老年性CRC早期诊断具有重要的应用价值。
简介:目的评价动脉内温热化疗并容式射频热疗治疗晚期胰腺癌的近期疗效。方法我科收治晚期胰腺癌11例,男8例,女3例。胰头癌8例,胰体尾癌3例,其中腺癌10例,囊腺癌1例。所有病人经Seldinger技术将导管选择性插至腹腔干和/或肠系膜上动脉,对胰体尾癌将导管插入脾动脉,留置并固定导管。将化疗药加热至43℃,进行动脉内热化疗,同时外施加容式射频热疗(预置温度43℃),共维持60min。间隔1—2mo重复治疗。介入治疗1—2mo,复查肿瘤标志物上腹部CT或MRI等影像学检查。同时评价病人临床受益反应(Clinicalbenefitresponse,CBR)。结果本组胰腺癌患者共接受21个疗程的介入性热化疗技术并容式射频热疗,其中1个疗程3例,2疗程6例,3个疗程2例,平均1.9个疗程。近期有效率54.5%,无1例肿瘤进展。3mo后,血肿瘤标志物CA199,CEA,以及胰腺癌相关抗原CA242检测均有明显下降(P<0.05)。临床受益率81.8%。结论介入性热化疗同时并以深部透热治疗不能手术根治的中晚期胰腺癌具有安全,有效,微创的优点。
简介:Objective:PyruvatekinasesM(PKM),includingthePKM1andPKM2isoforms,arecriticalfactorsinglucosemetabolism.PKM2promotesaerobicglycolysis,aphenomenonknownas"theWarburgeffect".ThepurposeofthisstudywastoidentifytherolesofPKM2inregulatingcellularmetabolism.Methods:TheCRISPR/Cas9systemwasusedtogeneratethePKM-knockoutcellmodeltoevaluatetheroleofPKMincellularmetabolism.LactatelevelsweremeasuredbytheVitrosLACslidemethodonanautoanalyzerandglucoselevelsweremeasuredbytheautoanalyzerAU5800.Themetabolismof13C6-glucoseor13C5-glutaminewasevaluatedbyliquidchromatography/massspectrometryanalyses.TheeffectsofPKMontumorgrowthweredetectedinvivoinatumor-bearingmousemodel.Results:WefoundthatbothPKM1andPKM2enabledaerobicglycolysis,butPKM2convertedglucosetolactatemuchmoreefficientlythanPKM1.Asaresult,PKM2reducedglucoselevelsreservedforintracellularutilization,particularlyfortheproductionofcitrate,andthusincreasedtheα-ketoglutarate/citrateratiotopromotethegenerationofglutamine-derivedacetylcoenzymeAthroughthereductivepathway.Furthermore,reductiveglutaminemetabolismfacilitatedcellproliferationunderhypoxiaconditions,whichsupportsinvivotumorgrowth.Inaddition,PKM-deletioninducedareverseWarburgeffectintumorassociatedstromalcells.Conclusions:PKM2playsacriticalroleinpromotingreductiveglutaminemetabolismandmaintainingprotonhomeostasis.ThisstudyishelpfultoincreasetheunderstandingofthephysiologicalroleofPKM2incancercells.
简介:Objective:TheresultsofapreviousstudyshowedthatacleardysregulationwasevidentintheglobalgeneexpressionoftheBCL11A-suppressedB-lymphomacells.Inthisstudy,thebonemorphogeneticproteinreceptor,typeII(BMPR2),E1Abindingproteinp300(EP300),transforminggrowthfactor-β2(TGFβ2),andtumornecrosisfactor,andalpha-inducedprotein3(TNFAIP3)geneexpressionpatternsinB-cellmalignancieswerestudied.Methods:TherelativeexpressionlevelsofBMPR2,EP300,TGFβ2,andTNFAIP3mRNAinB-lymphomacelllines,myeloidcelllines,aswellasincellsfromhealthyvolunteers,weredeterminedbyreal-timequantitativereversetranscriptpolymerasechainreaction(qRT-PCR)withSYBRGreenDye.Glyceraldehyde-3-phosphatedehydrogenase(GAPDH)wasusedasreference.Results:TheexpressionlevelofTGFβ2mRNAinB-lymphomacelllineswassignificantlyhigherthanthoseinthecellsfromthehealthycontrol(P<0.05).However,theexpressionlevelofTNFAIP3mRNAinB-malignantcellswassignificantlylowerthanthatofthehealthycontrol(P<0.05).TheexpressionlevelsofBMPR2andEP300mRNAshowednosignificantdifferencebetweenB-malignantcelllinesandthehealthygroup(P>0.05).InB-lymphomacelllines,correlationanalysesrevealedthattheexpressionofBMPR2andTNFAIP3(r=0.882,P=0.04)hadsignificantpositiverelation.TheexpressionlevelsofBMPR2,EP300,andTNFAIP3mRNAincelllinesfrommyeloidleukemiaweresignificantlylowerthanthoseinthecellsfromthehealthycontrol(P<0.05).TheexpressionlevelsofTGFβ2mRNAshowednosignificantdifferencebetweenmyeloidleukemiacelllinesandthehealthycontrolorB-malignantcelllines(P>0.05).TheexpressionlevelsofBMPR2,EP300,andTNFAIP3mRNAinB-lymphomacellsweresignificantlyhigherthanthoseofthemyeloidleukemiacells(P<0.05).Conclusion:DifferentexpressionpatternsofBMPR2,EP300,TGFβ2,andTNFAIP3genesinB-lymphomacellsexist.更多还原