简介:摘要丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)信号转导通路是调节细胞生长和存活的关键途径,异常的MAPK信号能促进癌症的发生发展,也决定了癌症的治疗反应。胃癌是中国常见消化道恶性肿瘤之一;近年来研究发现,在胃癌中存在诸多细胞信号传导通路的调控异常。细胞外信号调节激酶(extracellular signal-regulated kinase,ERK)信号通路是MAPK家族中的一条重要信号通路,该信号通路通过整合传递细胞外信号至细胞及其核内控制细胞的生长、凋亡和分化等。MAPK/ERK信号通路的异常激活可导致细胞丧失凋亡和分化能力,引发细胞异常增殖和恶性转化,促进胃癌的恶性表型;相反,阻断MAPK/ERK信号通路的相关组分则可逆转这一过程。本文结合国内外最新研究报道,对近年来在胃癌研究中发现MAPK/ERK信号通路的激活或失活在促进或抑制胃癌细胞恶性表型的重要作用及其相关的分子机制加以综述。
简介:目的研究ERK5在成骨细胞的表达及IL-6对其表达的调控,初步探讨ERK5在成骨细胞增殖分化中的作用。方法应用免疫印迹法检测人、大鼠、小鼠成骨细胞和MG-63细胞ERK5的表达;用IL-6作用于MG-63细胞,检测不同时间和剂量干预后ERK5和P-ERK5表达情况,构建ERK5siRNA和空白对照质粒,转染MG-63细胞,IL-6干预细胞后检测各组细胞增殖率、碱性磷酸酶活性和骨钙素表达的差异。结果在人、大鼠、小鼠成骨细胞和MG-63细胞中均有ERK5的表达;IL-6可呈时间、剂量依赖性的提高ERK5的磷酸化;与对照组相比,ERK5siRNA的成骨细胞IL-6刺激后增殖降低,骨钙素表达量下降且差异有统计学意义,而碱性磷酸酶活性的变化差异无显著性。结论成骨细胞ERK5信号通路的活性受IL-6调控,ERK5信号通路参与了成骨细胞的增殖分化。
简介:Objective:Theaimofthepresentstudywastoanalyzetheprognosticfactorsinpatientswithhepatoblastoma(HB)inoursinglecenterandtoevaluateperiostin(POSTN)expressioninHBanditsassociationwithclinicopathologicalvariables.Inaddition,theunderlyingmechanismofhowPOSTNpromotesHBprogressionwasdiscussed.Methods:POSTNexpressionwasinvestigatedinHBtumorsbyimmunohistochemistry(IHC),immunofluorescence(IF)andWesternblot(WB).TheassociationamongPOSTNexpression,clinicopathologicalfeaturesandoverallsurvival(OS)wasalsoevaluated.ThemigrationandadhesionabilityofHBcellsweremeasuredusingchemotaxisandcell-matrixadhesionassays,respectively.Epithelial-mesenchymaltransition(EMT)-associatedmarkersandactivationoftheERKpathwayweredetectedbyWB.Results:HBpatientshadpoorprognosiswhichdisplayedlymphnodemetastasis,vascularinvasion,POSTNandvimentinexpression.POSTNexpressionwasalsoassociatedwithlymphnodemetastasis.Furthermore,overexpressedPOSTNpromotedmigrationandtheadhesiveabilityofHBcellsinvitro.Inaddition,wedemonstratedthatPOSTNactivatedtheMAPK/ERKpathway,upregulatedtheexpressionofSnailanddecreasedtheexpressionofOVOL2.Finally,POSTNpromotedtheexpressionofEMT-associatedmarkers.Conclusions:POSTNmightmodulateEMTviatheERKsignalingpathway,therebypromotingcellularmigrationandinvasion.OurstudyalsosuggeststhatPOSTNmayserveasatherapeuticbiomarkerinHBpatients.
简介:摘要Ras/Raf/MEK/ERK信号传导通路系统与白血病治疗中对药物的敏感性和抗药性密切相关,Ras/Raf/MEK/ERK通路上游生长因子受体的变异和失调信号的传导均可导致与通路相关的重要遗传物质表达的变化。探讨Ras/Raf/MEK/ERK通路怎样的异常表达会导致白血病化疗耐药性和靶向治疗耐药性至关重要,如果能更好的控制Ras/Raf/MEK/ERK通路的表达可能会提高白血病治疗效果和改善患者的预后。
简介:摘要目的探讨Survivin和ERK在结直肠癌演变过程中的作用及其相关关系,为结直肠癌的早期诊断和治疗提供实验室依据和参考指标。方法应用免疫组织化学SP方法检测各组组织中Survivin和ERK二种蛋白的表达水平及相关分析。结果Survivin和ERK在结直肠切缘无瘤粘膜、腺瘤和腺癌组织中表达具有统计学显著性差异。在结直肠癌的患者中,Survivin蛋白在低分化组的表达显著高于高中分化组;在浆膜浸润组中的表达显著高于浆膜未浸润组;在淋巴结转移患者中显著高于无淋巴结转移的患者,且差异具有统计学意义(P<005);比较性别、年龄、肿瘤部位、瘤体大小之间的Survivin蛋白表达水平,未发现这些因素对Survivin蛋白表达的任何影响(P>005)。ERK蛋白在浆膜浸润组中的表达显著高于浆膜未浸润组;在淋巴结转移患者中显著高于无淋巴结转移的患者,且差异具有统计学意义(P<005);比较性别、年龄、肿瘤部位、瘤体大小和分化程度之间的Livin蛋白表达水平,未发现这些因素对Livin蛋白表达的任何影响(P>005)。Survivin和ERK在结直肠癌的表达之间有直线相关性(P<005),二者呈正相关。结论Survivin和ERK蛋白表达和结直肠癌发生发展相关。Survivin和ERK的表达与结直肠腺癌进展相关。Survivin和ERK在结直肠癌中的表达呈正相关。
简介:目的探讨在人骨髓间充质干细胞(hMSC)、小鼠前成骨细胞(MC3T3)和成牙本质细胞(MDPC-23)中,牙本质基质蛋白1(DMP1)是否通过激活丝裂原活化蛋白激酶(MAPK)-细胞外调节蛋白激酶(ERK)信号通路发挥作用.方法Westernblot检测不同时间点rDMP1C和rDMP1F蛋白处理hMSC、MC3T3-E1和MDPC-23后,对MAPK-ERK的激活情况;并检测使用MAPK抑制剂后,ERK的激活是否被抑制;细胞免疫荧光技术观察MAPK抑制剂抑制激活的ERK从胞浆向胞核的转位情况.Westernblot检测siRNA沉默Ras基因后,对rDMP1引起MAPK-ERK信号通路激活的影响.结果三种细胞中,rDMP1F和rDMP1C在5min~3h均可以激活MAPK-ERK通路,而总ERK在各时间点均无显著变化;rDMP1F激活信号通路的持续时间均明显长于rDMP1C;MAPK抑制剂处理组的p-ERK条带与rDMP1F/C处理组的p-ERK条带差别明显.hMSC中,rDMP1F激活MAPK信号通路持续时间要长于其在MC3T3和MDPC-23中的作用.细胞免疫荧光检测发现,MAPK抑制剂组可以阻断ERK向细胞核内的转位.MC3T3和MDPC-23在siRNA沉默Ras基因后,ERK的磷酸化水平与rDMP1F单独处理组相比显著降低.结论rDMP1C和rDMP1F都通过Ras激活MAPK-ERK信号通路,而rDMP1F比rDMP1C具有更强的信号功能.
简介:摘要目的观察督脉电针对脊髓损伤(SCI)小鼠MAPK/ERK1/2信号通路相关蛋白表达的影响。方法采用随机数字表法将64只C57BL/6小鼠分为假手术组、脊髓损伤组、督脉针刺组和督脉电针组。将脊髓损伤组、督脉针刺组及督脉电针组小鼠制成SCI动物模型。假手术组及脊髓损伤组小鼠制模后均未给予特殊处理,督脉针刺组小鼠于制模后次日给予单纯针刺治疗(取穴大椎、命门),督脉电针组小鼠于制模后次日给予电针治疗,取穴方法同督脉针刺组,电刺激设置疏密波(2/100 Hz),电刺激强度0.2 mA。于制模后3 d、7 d、14 d及28 d时分别采用免疫荧光、蛋白印迹法测定各组小鼠受损脊髓p-ERK1/2、p-Akt及髓鞘碱性蛋白(MBP)表达情况。结果与脊髓损伤组同期比较,督脉电针组在制模后3 d、14 d及28 d时,督脉针刺组在制模后28 d时其受损脊髓p-ERK1/2蛋白表达均显著增强(P<0.05);与督脉针刺组比较,督脉电针组在制模后3 d、14 d时其受损脊髓p-ERK1/2蛋白表达均明显增强(P<0.05)。督脉电针组在制模后14 d、28 d时,督脉针刺组在制模后28 d时其受损脊髓p-Akt蛋白表达均较脊髓损伤组显著增强(P<0.05),督脉针刺组在制模后3 d时其受损脊髓p-Akt蛋白表达较脊髓损伤组明显减弱(P<0.05)。督脉电针组在制模后3 d、7 d、14 d时,督脉针刺组在制模后3 d、7 d时其受损脊髓MBP蛋白表达均较脊髓损伤组明显增强(P<0.05)。结论督脉电针可促进SCI小鼠p-ERK1/2、p-Akt及MBP蛋白表达。
简介:AbstractBackground:Gastric cancer (GC) is one of the most globally prevalent cancers in the world. The pathogenesis of GC has not been fully elucidated, and there still lacks effective targeted therapeutics. The influence of altered kinesin superfamily protein 22 (KIF22) expression in GC progression is still unclearly. The aim of this study was to investigate the KIF22 effects on GC and related mechanisms.Methods:Gastric carcinoma tissues and matching non-cancerous tissues were collected from patients with GC who have accepted a radical gastrectomy in Lanzhou University Second Hospital from May 2013 to December 2014. The expression of KIF22 was examined in GC of 67 patients and 20 para-carcinoma tissues by immunochemical staining. The relationship between the expression of KIF22 and clinicopathologic characteristics was next investigated in the remaining 52 patients except for 15 patients who did not complete follow-up for 5 years. Cell viability was performed via 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) test and colony formation assay in the MGC-803 and BGC-823 GC cells. Cell scratch and trans-well invasion assay was performed to assess migration ability in the MGC-803 and BGC-823 GC cells. Gene set enrichment analysis (GSEA) pathway enrichment analysis was performed to explore the potential functions. Cell cycle was detected by flow cytometry. In addition, the two GC cell lines were used to elucidate the underlying mechanism of KIF22 in GC in vitro via assessing the effects on mitogenactivated protein kinase and extracellular regulated protein kinases (MAPK/ERK) signal transduction pathway-related expressions by Western blotting assays. The differences were compared by t tests, one-way analysis of variance, and Chi-squared tests.Results:The study showed that KIF22 was up-regulated in GC, and KIF22 high expression was significantly related to differentiation degree (χ2 = 12.842, P = 0.002) and poorly overall survivals. GSEA pathway enrichment analysis showed that KIF22 was correlated with the cell cycle. Silence of KIF22 decreased the ability of the proliferation and migration in gastric cells, induced G1/S phase cell cycle arrest via regulating the MAPK-ERK pathways.Conclusions:KIF22 protein level was negatively correlated with prognosis. KIF22 knockdown might inhibit proliferation and metastasis of GC cells via the MAPK-ERK signaling pathway.
简介:由二个原子形成的双原子分子极性的判断学生容易掌握,而多原子分子极性的判断学生往往束手无策。本人根据自己的教学体会认为,用以下三种方法可以较好地解决这一问题。一、根据中心元素的化合价进行判断在共价化合物中,若中心元素的化合价处于最高价,说明在分子中中心原子的最外层电子全部参与成键。按价电子对互斥理论,分子具有理想的几何构型,按照理想几何构型分子内部具有对称性,其偶极矩为零。即:分子为非极性分子。当中心元素的化合价未处于最高时,在分子中中心原子除了参与成键的电子外,尚存在孤对电子,由于孤对电子与成键电子间的相互作用,使分子构型发生扭曲而失去对称性成为极性分子。据此方法学生不难判断出:
简介:目的:探讨ERK1/2信号通路阻断剂PD98059对Ras、B型丝/苏氨酸蛋白激酶(BRaf)、丝裂原活化蛋白激酶(MEK)、细胞信号调节激酶1/2(ERK1/2)表达的影响,以期为慢性高原病(chronicmountainsickness,CMS)的基础研究和临床治疗探讨新的途径。方法:选取CMS患者16例,取骨髓液分离单个核细胞,以CD71与CD235a抗体磁珠分选阳性细胞,将细胞分为5组:空白对照组、DMSO溶剂组及PD980595、10和20μmol/L组。在低氧条件下培养72h,应用ELISA法测定培养骨髓有核红细胞上清液中Ras-GTP水平,RT-PCR法测定骨髓有核红细胞中BRaf、MEK、ERK1/2mRNA的表达,Westernblot方法检测骨髓有核红细胞中p-BRaf、p-MEK、pERK1/2蛋白表达。结果:PD98059对各组Ras-GTP的水平无明显影响(P=0.798)。溶剂组与空白对照组相比,BRaf、MEKmRNA的表达水平差异无统计学意义(P=0.826、P=0.298)。与PD9805920mol/L比较,其余4组ERK1/2mRNA的表达水平差异有统计学意义(P=0.001、P=0.002)。溶剂组与空白对照组相比,p-BRaf、p-MEK蛋白的表达差异无统计学意义(P=0.370、P=0.351)。与PD9805920mol/L比较,其余4组p-ERK1/2蛋白水平差异有统计学意义(P〈0.001、P〈0.007)。结论:PD98059能下调骨髓有核红细胞ERK1/2mRNA及p-ERK1/2蛋白的表达。Ras/Raf/MEK/ERK1/2通路是调控CMS骨髓有核红细胞的主要信号传导途径,可能参与了慢性高原病的发病过程。