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1、大鼠增殖抑制基因抑制大鼠膠質(zhì)瘤細(xì)胞的增殖及其機(jī)制高鵬1,2，蔣樹財(cái)1，郭暉3，楊強(qiáng)1，方志豪1，趙巍1、4，沈冰2△(1.寧夏醫(yī)科大學(xué)，銀川750004；2.寧夏醫(yī)科大學(xué)總醫(yī)院神經(jīng)外科，銀川750004；3.濰坊市人民醫(yī)院神經(jīng)外科，濰坊261000；4.寧夏醫(yī)科大學(xué)醫(yī)學(xué)科學(xué)技術(shù)研究中心，銀川750004)【摘 要】目的：觀察腺病毒介導(dǎo)的大鼠增殖抑制基因(rathyperplasiasuppressorgene,rHSG)在大鼠膠質(zhì)瘤細(xì)胞(C6)中的表達(dá)及其對大鼠膠質(zhì)瘤細(xì)胞增殖的影響，并探討其作用
2、的機(jī)制。方法：用含有rHSG基因的重組腺病毒載體(Adv-rHSG-GFP)轉(zhuǎn)染C6細(xì)胞，采用Westernblot法和免疫細(xì)胞化學(xué)法檢測rHSG蛋白的表達(dá)；使用流式細(xì)胞儀分析細(xì)胞周期，并結(jié)合MTT、細(xì)胞計(jì)數(shù)法和平板克隆形成實(shí)驗(yàn)觀察rHSG對細(xì)胞增殖的影響；應(yīng)用Westernblot法檢測增殖細(xì)胞核抗原(proliferatingcellnuclearantigen,PCNA)和細(xì)胞周期調(diào)控蛋白p27Kip1、p21Cip1的蛋白表達(dá)變化。結(jié)果:Adv-rHSG-GFP轉(zhuǎn)染組C6細(xì)胞中：rHSG
3、蛋白的相對表達(dá)量均明顯高于未轉(zhuǎn)染組和Adv-GFP轉(zhuǎn)染組(P<0.01)；流式細(xì)胞儀分析結(jié)果表明細(xì)胞周期被阻滯在G0/G1期，且MTT、細(xì)胞計(jì)數(shù)和平板克隆形成實(shí)驗(yàn)顯示大鼠膠質(zhì)瘤細(xì)胞增殖受到明顯抑制(P<0.01)；周期調(diào)控蛋白p27Kip1、p21Cip1表達(dá)升高，PCNA表達(dá)降低。結(jié)論：rHSG基因過表達(dá)能夠抑制大鼠膠質(zhì)瘤細(xì)胞增殖，其機(jī)制可能是通過參與細(xì)胞周期調(diào)節(jié)發(fā)揮作用?！娟P(guān)鍵詞】神經(jīng)膠質(zhì)瘤；腺病毒；大鼠增殖抑制基因；細(xì)胞，C6【中圖分類號】R739.41 【文獻(xiàn)標(biāo)識碼】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
