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1、萬方數(shù)據(jù)·20·．口腔頜面腫瘤防治．蛋白多糖與涎腺腺樣囊性癌增殖的關(guān)系石宏王潔董福生王旭李荷香【摘要】目的探討蛋白多糖(proteoglycans)合成的阻抑對人涎腺腺樣囊性癌ACC．M細胞增殖的影響。方法構(gòu)建靶向人木糖基轉(zhuǎn)移酶-I(xylosyltransferase—I，XT—I)基因的短發(fā)卡狀RNA(shorthairpinRNA，shRNA)真核表達載體shRNA—WJ3，轉(zhuǎn)染人涎腺腺樣囊性癌高轉(zhuǎn)移株ACC．M細胞，通過G418篩選穩(wěn)定轉(zhuǎn)染的細胞，采用實時定量聚合酶鏈反應和蛋白質(zhì)印跡法測定shRNA．WJ3的基因沉默效果；檢測各組細胞蛋白多糖合成分泌水平的變化
2、。采用甲基嚷唑基四唑法(M1Tr)檢測細胞增殖情況；流式細胞術(shù)測定細胞周期改變。結(jié)果shRNA-WJ3轉(zhuǎn)染ACC．，M細胞后，細胞中XT．I基因的mRNA和蛋白表達抑制率分別為83．70％和79．60％；穩(wěn)定轉(zhuǎn)染shRNA-WJ3的ACC．M．WJ3細胞蛋白多糖分泌顯著降低(P<0．01)，抑制率為49．7l％一54．59％。MTr法檢測結(jié)果表明：蛋白多糖合成阻抑后，ACC-M細胞增殖活性明顯降低；ACC—M-WJ3細胞與對照組細胞相比：s期細胞數(shù)日減少；G。一Go期細胞數(shù)目增加，差異有統(tǒng)計學意義(P<0．01)。結(jié)論蛋白多糖合成分泌的阻抑能有效抑制ACC—M細胞的增
3、殖?！娟P(guān)鍵詞】癌，腺樣囊性；涎腺；蛋白多糖RelationshipbetweenproteoglycansandproliferationofhumansalivaryadencidcysticcarcinomasH
4、Hong。WAl、Gj沈’，DONGFu—sheng，WANGXu．uHe—xiang．‘DepartmentofOralPathology。CollegeofStomatology，脅beiMedicalUniversity．Sh“iazhuang050017，ChinaCorrespondingauthor：WANGJie，Email：wangiie
5、phd@163．COIn，Td：0086—3l1—86265780【Abstract】ObjectiveToinvestigatetheeffectofdown·regulatedproteoglycansontheproliferationofhumansalivaryadenoidcysticcarcinoma(SACC)．MethodsTheshorthairpinRNA(shRNA)plasmidsilencinghumanxylosyhransferase—I(XT-I)geneWflftconstructedandnamedshRNA．WJ3．Adenoi
6、dcysticcarcinomacellswithhighmetastatictendency(ACC．M)weretransfectedbyshRNA．WJ3．TheplasmidshRNA—HKnottargetinganyhumangenewastransfectedintoACC．Mcellsused鼬negativecontroLAfter48hoftransfection．t}IepositivecellswerescreenedbyC-418toisolatethestabletransfectedceils．Real-timePCRandWestern
7、blottingwereusedtotestthegenesilence。andtheproteoglycanscontentsofthecellsweredetected．ThestablecelllinesilencedXT-IWasnamedACC-M—WJ3．M33'assaywa$performedtodetectthecellproliferation．ThecellcycleWaSanalyzedbyflowcytometry．ResultsShRNA．WJ3showedpowerfulRNAinterferenceandgenesilenceofXT-
8、I．Theinhibitionratewas83．70％ofmRNAexpressionand79．60％ofproteinexpressionrespectively．111econtentofproteoglycansinACC-M—WJ3Wasdown-regulatedby49．71％-54．59％．TheresultsofMTrassayshowedthattheeellgrowthWasinhibitedsignificantly．SphrasedecreasedandGI—GophraseincreasedingroupACC—M-WJ
