简介:摘要目的研究泛醌氧化还原酶复合物组装因子4(NDUFAF4)的表达与肝癌患者临床预后的相关性,探究NDUFAF4对人肝癌细胞系的辐射敏感性的影响及其机制。方法通过数据库及肝癌组织样本探究NDUFAF4在肝癌中的表达及其表达水平与临床预后的相关性。通过脂质体将敲减NDUFAF4基因的质粒si-NDUFAF4转入HepG2和Huh7细胞,克隆形成实验检测辐射敏感性。同时通过裸鼠开展荷瘤实验,免疫荧光法检测β联蛋白(β-catenin),蛋白质印迹法检测上皮钙黏素(E-cadherin)和神经钙黏素(N-cadherin)的蛋白表达。结果生物信息学分析结果证实,NDUFAF4在肝癌组织中显著高表达,其表达水平越高,患者预后越差(P<0.05)。NDUFAF4在肝癌组织中的表达水平显著高于癌旁组织;克隆形成实验证实,敲减NDUFAF4显著降低肝癌细胞的生存率(P<0.01);体内实验证实,敲减NDUFAF4可以减慢癌细胞增殖,减少β-catenin及Ki-67的表达。敲减NDUFAF4显著减少肝癌细胞系核内β-catenin的蛋白水平,提示NDUFAF4可以激活Wnt/β-catenin通路。敲减NDUFAF4显著上调E-cadherin和下调N-cadherin的蛋白表达水平。结论敲减NDUFAF4可以通过抑制Wnt/β-catenin通路增强人肝癌细胞系的辐射敏感性,并且与临床预后紧密相关,或可为克服肝癌的辐射抗性提供新的靶点。
简介:AbstractBackground:Histone deacetylase 4 (HDAC4) regulates chondrocyte hypertrophy and bone formation. The aim of the present study was to explore the effects of HDAC4 on Interleukin 1 beta (IL-1β)-induced chondrocyte extracellular matrix degradation and whether it is regulated through the WNT family member 3A (WNT3A)/β-catenin signaling pathway.Methods:Primary chondrocytes (CC) and human chondrosarcoma cells (SW1353 cells) were treated with IL-1β and the level of HDAC4 was assayed using Western blotting. Then, HDAC4 expression in the SW1353 cells was silenced using small interfering RNA to detect the effect of HDAC4 knockdown on the levels of matrix metalloproteinase 3 (MMP3) and MMP13 induced by IL-1β. After transfection with HDAC4 plasmids, the overexpression efficiency was examined using Real-time quantitative polymerase chain reaction (qRT-PCR) and the levels of MMP3 and MMP13 were assayed using Western blotting. After incubation with IL-1β, the translocation of β-catenin into the nucleus was observed using immunofluorescence staining in SW1353 cells to investigate the activation of the WNT3A/β-catenin signaling pathway. Finally, treatment with WNT3A and transfection with glycogen synthase kinase 3 beta (GSK3β) plasmids were assessed for their effects on HDAC4 levels using Western blotting.Results:IL-1β downregulated HDAC4 levels in chondrocytes and SW1353 cells. Furthermore, HDAC4 knockdown increased the levels of MMP3 and MMP13, which contributed to the degradation of the extracellular matrix. Overexpression of HDAC4 inhibited IL-1β-induced increases in MMP3 and MMP13. IL-1β upregulated the levels of WNT3A, and WNT3A reduced HDAC4 levels in SW1353 cells. GSK-3β rescued IL-1β-induced downregulation of HDAC4 in SW1353 cells.Conclusion:HDAC4 exerted an inhibitory effect on IL-1β-induced extracellular matrix degradation and was regulated partially by the WNT3A/β-catenin signaling pathway.
简介:摘要目的临床观察β-catenin、Oct-4以及Wnt1对胃癌患者检查的意义。方法按入院时间选取90例我院2014.3~2015.4期间经诊断为胃癌的患者,取其癌组织作为观察组,同时取患者癌旁正常组织作为对照组,均使用β-catenin、Oct-4以及Wnt1进行临床检查,对各组阳性率进行比较分析。结果观察组较对照组各指标阳性率高,P<0.05;抗原阳性反应与肿瘤大小、分化程度有关,P<0.05,与患者性别无关,P>0.05。结论临床采用β-catenin、Oct-4以及Wnt1对患者进行检查可有效诊断患者病情以及癌细胞分化程度。
简介:摘要目的探讨Wnt/β-连环链蛋白(catenin)/TCF-4通路在肾癌细胞中的作用,并初步分析其可能的机制。方法分别构建β-catenin sh和TCF-4△Nsh的真核表达载体,并分别转染肾癌786-O细胞,采用CCK8检测转染后细胞的增殖能力,吖啶橙-溴乙锭(AO-EB)染色法检测转染后细胞死亡情况,Western印迹检测转染组、空转组以及空白组细胞TCF-4、Bcl-2、bax、Caspase-3的表达情况。结果转染β-catenin siRNA病毒后,细胞的增殖能力较空转组明显降低(0.443±0.145与0.910±0.721),细胞死亡率较空转组明显增加(16.38±5.32与6.61±1.04),TCF-4的表达受抑制、凋亡蛋白Caspase 3、bax表达增加、抗凋亡蛋白Bcl-2表达减低(均P<0.05)。转染TCF-4 siRNA病毒后,细胞的增殖能力较对照组明显降低,细胞死亡率较对照组明显增加,TCF-4的表达受抑制导致凋亡蛋白Caspase 3、bax表达增加,抗凋亡蛋白Bcl-2表达减低(均P<0.05)。结论Wnt/β-catenin/TCF-4通路在786-O肾癌细胞中具有可调节肾癌细胞的增殖和凋亡的作用,其机制可能为通过调节其下游的凋亡蛋白Caspase 3、bax和抗凋亡蛋白Bcl-2的表达水平来实现。
简介:Irradiationfromdiversesourcesisubiquitousandcloselyassociatedwithhumanactivities.Radiationtherapy(RT),animportantcomponentofmultipleradiationorigins,isacommontherapeuticmodalityforcancer.Moreimportantly,RTprovidessignificantcontributiontooncotherapybykillingtumorcells.However,duringthecourseoftherapy,irradiationofnormaltissuescanresultinawiderangeofsideeffects,includingself-limitedacutetoxicities,mildchronicsymptoms,orsevereorgandysfunction.Althoughnumerouspromisingradioprotectiveagentshaveemerged,onlyafewhavesuccessfullyenteredthemarketbecauseofvariouslimitations.Atpresent,thewidelyacceptedhypothesisforprotectionagainstradiation-causedinjuryinvolvestheWntcanonicalpathway.ActivatingtheWnt/β-cateninsignalingpathwaymayprotectthesalivarygland,oralmucosa,andgastrointestinalepitheliumfromradiationdamage.Theunderlyingmechanismsincludeinhibitingapoptosisandpreservingnormaltissuefunctions.However,aberrantWntsignalingunderliesawiderangeofpathologiesinhumans,anditsvariouscomponentscontributetocancer.Moreover,studieshavesuggestedthatWnt/β-cateninsignalingmayleadtoradioresistanceofcancerstemcell.ThesefactsmarkedlycomplicateanydefinitionoftheexactfunctionoftheWntpathway.
简介:Masterdevelopmentalpathways,suchasNotch,Wnt,andHedgehog,aresignalingsystemsthatcontrolproliferation,celldeath,motility,migration,andstemness.Thesesystemsarenotonlycommonlyactivatedinmanysolidtumors,wheretheydriveorcontributetocancerinitiation,butalsoinprimaryandmetastatictumordevelopment.Thereactivationofdevelopmentalpathwaysincancerstromafavorsthedevelopmentofcancerstemcellsandallowstheirmaintenance,indicatingthesesignalingpathwaysasparticularlyattractivetargetsforefficientanticancertherapies,especiallyinadvancedprimarytumorsandmetastaticcancers.Metastasisistheworstfeatureofcancerdevelopment.Thisfeatureresultsfromacascadeofeventsemergingfromthehijackingofepithelial-mesenchymaltransition,angiogenesis,migration,andinvasionbytransformingcellsandisassociatedwithpoorsurvival,drugresistance,andtumorrelapse.Inthepresentreview,wesummarizeanddiscussexperimentaldatasuggestingpivotalrolesfordevelopmentalpathwaysincancerdevelopmentandmetastasis,consideringthetherapeuticpotential.EmergingtargetedantimetastatictherapiesbasedonNotch,Wnt,andHedgehogpathwaysarealsodiscussed.
简介:摘要Wnt信号通路在生物发育、细胞转运及凋亡等过程中发挥非常重要的作用,其异常活化与胃癌密切相关。本文对经典Wnt信号传导通路中各组成部分、相互作用,与胃癌发生、发展的关系作一总结,并分析Wint信号通路及其受体的研究进展。
简介:摘要目的探讨受体相互作用蛋白激酶4(RIPK4)在乳腺癌发生、发展中的作用及其机制。方法收集郑州大学第一附属医院乳腺外科2017年10月至2018年7月手术切除的乳腺癌和癌旁正常组织96例,采用免疫组织化学方法检测RIPK4蛋白在96例乳腺癌和癌旁正常组织中的表达及其与乳腺癌临床病理特征间的关系;靶向RIPK4的小干扰RNA(siRNA)瞬时转染乳腺癌MCF-7、MDA-MB-231细胞,同时以转染阴性对照siRNA和空白细胞为对照,共分为3组:RIPK4 siRNA转染组、空白细胞组和阴性对照siRNA转染组。细胞计数试剂盒(CCK-8)试验和侵袭小室试验检测抑制RIPK4蛋白表达对MCF-7、MDA-MB-231细胞增殖和侵袭转移能力的影响,蛋白质印迹法(Western blot)检测抑制RIPK4蛋白表达对Wnt/β-连环蛋白(β-catenin)信号通路的影响。采用方差分析进行组间比较,采用t检验进行两两比较。结果免疫组织化学结果显示RIPK4蛋白在乳腺癌组织中的表达明显增高(71.9%,69/96),与癌旁正常组织比较(21.9%,21/96),差异有统计学意义(χ2=48.188,P<0.01);且RIPK4蛋白异常高表达与乳腺癌患者的TNM分期、组织学类型及淋巴结转移明显相关(χ2=17.524、40.697、9.458,P<0.05);靶向RIPK4的siRNA瞬时转染MCF-7、MDA-MB-231细胞后,CCK-8试验和侵袭小室试验结果显示,MCF-7、MDA-MB-231细胞的增殖和侵袭转移能力降低;进一步试验表明RIPK4 siRNA抑制MCF-7、MDA-MB-231细胞中RIPK4蛋白的表达后,Wnt/β-catenin信号通路中β-catenin表达随之下降,上皮-间充质转化(EMT)的相关蛋白上皮型钙黏蛋白(E-cadherin)表达增高而波形蛋白(Vimentin)表达下降。结论RIPK4蛋白在乳腺癌组织中异常高表达,抑制RIPK4蛋白的表达有可能通过抑制Wnt/β-catenin信号通路相关的EMT的发生降低乳腺癌细胞的增殖、侵袭转移能力。
简介:摘要目的观察细胞外因子wnt-1和wnt-3a及胞质蛋白β-catenin在蛋白水平和基因的表达水平,讨论其在宫颈癌的发生和发展中的作用,为宫颈癌发生和发展的病理机理提供理论依据。方法利用免疫组化法检验正常宫颈组织12例,宫颈上皮内瘤变组织18例,宫颈鳞癌组织20例中wnt-1、wnt-3a和β-catenin蛋白的表达,并对检测的数据进行数据统计学分析。结果wnt-3a在正常的宫颈组织、宫颈上皮内瘤变和宫颈鳞癌组织的阳性表达率分别为10%、37.5%和39.3%,三组的数据进行比较可以看到具有显著性差异(p<0.05)。结论wnt与β-catenin的表达异常与宫颈癌发生和发展具有密切的关系,有可能是进行宫颈癌早期诊断的附加标准,而wnt-3a蛋白与β-catenin蛋白进行联合检测可能会更精确的对宫颈癌早期的早期诊断和对其病理恶性程度进行判断。
简介:摘要目的探讨CXXC型锌指蛋白4基因(CXXC4)对胃癌细胞增殖和凋亡的影响及其机制。方法采用甲基化特异性聚合酶链反应(PCR)检测江苏省苏北人民医院2015年1月至2019年12月诊治的100例胃癌患者(胃癌组)和50例胃良性疾病患者(良性组)CXXC4基因甲基化并分析其与临床病理因素关系。设计合成靶向CXXC4基因的甲基化寡核苷酸转染至SGC7901胃癌细胞(转染甲基化寡核苷酸组,MON组),选择未转染甲基化寡核苷酸细胞为对照(未转染寡核苷酸,CON组),单因素方差分析比较两组细胞CXXC4基因甲基化及信使核糖核酸(mRNA)表达、吸光度(A)值、细胞周期及凋亡、果蝇无翅基因(Wnt)、β-连环蛋白(β-catenin)、细胞周期蛋白D1(Cyclin D1)和c-Myc基因mRNA表达。两组间比较采用t或χ2检验,多组间比较采用单因素方差分析,采用SNK-q法分别比较组间差异。结果胃癌组患者CXXC4基因甲基化率显著高于CON组(67.0%比14.0%,χ2=35.371,P<0.01),差异有统计学意义。SGC7901胃癌细胞中CXXC4为未甲基化状态。胃癌组患者CXXC4基因甲基化率在分化程度、肿瘤T分期、淋巴结转移及肿瘤分期方面的差异有统计学意义(χ2=4.626、4.075、5.294、5.619,P值均<0.05)。MON组SGC7901胃癌细胞第1~6天A值、S期、G2/M期、增殖指数、Wnt、β-catenin、Cyclin D1和c-Myc基因mRNA相对表达量显著高于CON组(t=7.324、9.238、8.789、11.233、8.018、10.383、27.899、34.461、21.432、20.137、31.654、27.439、36.872,P<0.01),差异有统计学意义,CXXC4基因mRNA表达、G0/G1期和凋亡率显著低于CON组(t=55.637、45.256、32.009,P<0.01),差异有统计学意义。结论CXXC4基因甲基化与胃癌发病机制及临床病理因素有相关。CXXC4基因甲基化上调Wnt/β-catenin信号通路促进胃癌细胞增殖并抑制凋亡。
简介:AbstractGlioblastoma is the most common form of primary brain tumor. Glioblastoma stem cells play an important role in tumor formation by activation of several signaling pathways. Wnt signaling pathway is one such important pathway which helps cellular differentiation to promote tumor formation in the brain. Glioblastoma remains to be a highly destructive type of tumor despite availability of treatment strategies like surgery, chemotherapy, and radiation. Advances in the field of cancer biology have revolutionized therapy by allowing targeting of tumor-specific molecular deregulation. In this review, we discuss about the significance of glioblastoma stem cells in cancer progression through Wnt signaling pathway and highlight the clinical targets being potentially considered for therapy in glioblastoma.
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简介:摘要Wnt/β-catcnin信号通路是Wnt信号传导通路中最为经典的信号通路。它是调节软骨代谢的重要途径。Wnt蛋白被证实在软骨基质合成和转归,骨质形成和转归中发挥着重要作用。氧化应激所致的细胞老化过程中,p53/p21在细胞老化调控通路中发挥着核心作用。p53/p21通路在Wnt信号激活关节软骨促间充质祖细胞老化中起介导作用。本文就Wnt信号通路与软骨细胞老化的研究进展展开综述。