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1、大鼠增殖抑制基因抑制大鼠胶质瘤细胞的增殖及其机制高鹏1,2,蒋树财1,郭晖3,杨强1,方志豪1,赵巍1、4,沈冰2△(1.宁夏医科大学,银川750004;2.宁夏医科大学总医院神经外科,银川750004;3.潍坊市人民医院神经外科,潍坊261000;4.宁夏医科大学医学科学技术研究中心,银川750004)【摘 要】目的:观察腺病毒介导的大鼠增殖抑制基因(rathyperplasiasuppressorgene,rHSG)在大鼠胶质瘤细胞(C6)中的表达及其对大鼠胶质瘤细胞增殖的影响,并探讨其作用
2、的机制。方法:用含有rHSG基因的重组腺病毒载体(Adv-rHSG-GFP)转染C6细胞,采用Westernblot法和免疫细胞化学法检测rHSG蛋白的表达;使用流式细胞仪分析细胞周期,并结合MTT、细胞计数法和平板克隆形成实验观察rHSG对细胞增殖的影响;应用Westernblot法检测增殖细胞核抗原(proliferatingcellnuclearantigen,PCNA)和细胞周期调控蛋白p27Kip1、p21Cip1的蛋白表达变化。结果:Adv-rHSG-GFP转染组C6细胞中:rHSG
3、蛋白的相对表达量均明显高于未转染组和Adv-GFP转染组(P<0.01);流式细胞仪分析结果表明细胞周期被阻滞在G0/G1期,且MTT、细胞计数和平板克隆形成实验显示大鼠胶质瘤细胞增殖受到明显抑制(P<0.01);周期调控蛋白p27Kip1、p21Cip1表达升高,PCNA表达降低。结论:rHSG基因过表达能够抑制大鼠胶质瘤细胞增殖,其机制可能是通过参与细胞周期调节发挥作用。【关键词】神经胶质瘤;腺病毒;大鼠增殖抑制基因;细胞,C6【中图分类号】R739.41 【文献标识码】AExpressio
4、nandmechanismofrathyperplasiasuppressorgenesuppressinggrowthofratc6gliomacellsGAOPeng1,2,JIANGShu-cai1,GUOHui3,YANGQiang1,FANGZhi-hao1,ZHAOWei1,4,SHENBing2△(1.NingxiaMedicalUniversity,Yinchuan750004;2.DepartmentofNeurosurgery,GeneralHospitalofNingxia
5、MedicalUniversity,Yinchuan750004;3.DepartmentofNeurosurgery,WeifangPeople’sHospital,Weifang261000;4.MedicalSci-TechResearchCenterofNingxiaMedicalUniversity,Yinchuan750004;△Correspondingauthor)[Abstract]Objective:Thepurposeofthisstudyistoinvestigateth
6、eexpressionofrathyperplasiasuppressorgene(rHSG)inratgliomacellaswellaseffectofrHSGonratgliomacellproliferation,andexplorethemechanism.Methods:Theratgliomacells(C6cells)wereinfectedwithrecombinantadenovirusvector containingrHSG(Adv-rHSG-GFP).Westernbl
7、otandimmunohistochemicalmethod wereappliedfordeterminingtheexpressionofrHSGprotein;theeffectofrHSGoncellcyclewasanalyzedby flowcytometer,togetherwithMTT, cytometryandplatecloningformationmethodwereusedforidentifyingtheeffectofrHSGoncellproliferation;
8、theexpressionofproliferatingcellnuclearantigen(PCNA), cellcycleregulatoryproteinsp27Kip1andp21Cip1weredeterminedbywesternblot.Results:ItisobservedthatinC6cellsofgroupinfected withAdv-rHSG-GFP,theproteinexpressionlevelofrHSGwasmorethangroupofuninfecte