蛋白多糖与涎腺腺样囊性癌增殖的关系

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1、万方数据·20·．口腔颌面肿瘤防治．蛋白多糖与涎腺腺样囊性癌增殖的关系石宏王洁董福生王旭李荷香【摘要】目的探讨蛋白多糖(proteoglycans)合成的阻抑对人涎腺腺样囊性癌ACC．M细胞增殖的影响。方法构建靶向人木糖基转移酶-I(xylosyltransferase—I，XT—I)基因的短发卡状RNA(shorthairpinRNA，shRNA)真核表达载体shRNA—WJ3，转染人涎腺腺样囊性癌高转移株ACC．M细胞，通过G418筛选稳定转染的细胞，采用实时定量聚合酶链反应和蛋白质印迹法测定shRNA．WJ3的基因沉默效果；检测各组细胞蛋白多糖合成分泌水平的变化

2、。采用甲基嚷唑基四唑法(M1Tr)检测细胞增殖情况；流式细胞术测定细胞周期改变。结果shRNA-WJ3转染ACC．，M细胞后，细胞中XT．I基因的mRNA和蛋白表达抑制率分别为83．70％和79．60％；稳定转染shRNA-WJ3的ACC．M．WJ3细胞蛋白多糖分泌显著降低(P<0．01)，抑制率为49．7l％一54．59％。MTr法检测结果表明：蛋白多糖合成阻抑后，ACC-M细胞增殖活性明显降低；ACC—M-WJ3细胞与对照组细胞相比：s期细胞数日减少；G。一Go期细胞数目增加，差异有统计学意义(P<0．01)。结论蛋白多糖合成分泌的阻抑能有效抑制ACC—M细胞的增

3、殖。【关键词】癌，腺样囊性；涎腺；蛋白多糖RelationshipbetweenproteoglycansandproliferationofhumansalivaryadencidcysticcarcinomasH

4、Hong。WAl、Gj沈’，DONGFu—sheng，WANGXu．uHe—xiang．‘DepartmentofOralPathology。CollegeofStomatology，胁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