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ID:34064447
大小:4.81 MB
页数:44页
时间:2019-03-03
《沉默高尔基体α-甘露糖苷酶ⅱ对胃部细胞黏附的影响》由会员上传分享,免费在线阅读,更多相关内容在学术论文-天天文库。
1、重庆医科大学硕士学位论文沉默高尔基体α--甘露糖苷酶Ⅱ对胃部细胞黏附的影响姓名:杨紫汐申请学位级别:硕士专业:病理学与病理生理学指导教师:易永芬201205重庆医科大学硕士研究生学位论文沉默高尔基体Q一甘露糖苷酶Ⅱ对胃癌细胞黏附的影响摘要目的:探讨沉默高尔基体Q一甘露糖苷酶II(GMII)对人胃癌细胞株BGC一823黏附活性的影响及其可能相关机制。方法:通过脂质体Lipofectamine2000将GMII—shRNA真核表达质粒载体转染至人胃癌细胞株BGC一823,利用免疫荧光观察其转染效率。用RT—PCR、Westernblot方法检测转染细胞GMlI、CD44及E—selectin
2、mRNA和蛋白表达水平的变化。用MTT比色试验测定胃癌细胞在细胞外c基质上黏附能力的变化。通过胃癌细胞对RoseBengal的摄取测定其与血管内皮细胞黏附能力的变化。结果:成功将GMII—shRNA真核表达质粒载体转染至人胃癌细胞株BGC一823。转染后胃癌细胞GMII及CD44的mRNA和蛋白表达水平明显低于对照组(P3、细胞与血管内皮细胞黏附试验结果显示,转染GMII—shRNA真核表达质粒组细胞的吸光度A值明显低于转染空载体组和空白对照组(P4、,黏附EFFECToFRNAINTERFERENCETARGETINGGMⅡoNADHESIoNoFGASTRICCELLSABSTRACTObjective:TranSfectedthes删AplasmidofhumanG0培i弘lna彻osldaseII(GMII)t0gastriccancercenIineBGC一823,identi匆itsexpressioniIlBGC-823,andinvestigateitse腩ctonadhesionofthecells·Memods:Trans胁edtheGMII.shRNAplasmidtoBGC-823with1ipofe咖1me'5、andthetransfectionefficiencyⅥ娜provedbyFluoresc饥ceMicroscopy.RT.PCRwe‘eusedt0detefmmethemRNAexpressioIlsofGMⅡ、CD44andE。seledmaRertransfection.wresternblot、ⅣereuSedt0detecttheprotemeXpressionsofGMII、CD44andE.selectiIlaRer仃ansfection.AMTTassayandanadhesionteStwereusedt0一⋯一dete嗍inetheB(祀.823Cell’sadh6、esiVeabilityaRer仃aIlSfectiorLResults:TheGMII.shI矾AplasmidwassuCcessfullytmllsfected.Rr-PCRandwest锄blota11alysisindicatedthatmRNAandproteineXpressionofGM11wasdown-regulatedsignifiCantlyi11gastriccancercelllineBGC-823aRertms触ion(P<0.05).MrrandadhesionteStshowedthatthegroupwithGMII。s删Aledtoa10weradh7、esiveabil“ytIlanothers(P<0.05).Comparet0theC0ntrol鲈oup,‘hegrouptransf&tedwithGMII.shI矾Ashowed0bViously10w可CD44n‘武NAanadproteinexpreSsion(P<0.05).AmdE—selectill’smI冰Aandprotein能presslonwaSn0‘changeobViously.Conclusion:T
3、细胞与血管内皮细胞黏附试验结果显示,转染GMII—shRNA真核表达质粒组细胞的吸光度A值明显低于转染空载体组和空白对照组(P4、,黏附EFFECToFRNAINTERFERENCETARGETINGGMⅡoNADHESIoNoFGASTRICCELLSABSTRACTObjective:TranSfectedthes删AplasmidofhumanG0培i弘lna彻osldaseII(GMII)t0gastriccancercenIineBGC一823,identi匆itsexpressioniIlBGC-823,andinvestigateitse腩ctonadhesionofthecells·Memods:Trans胁edtheGMII.shRNAplasmidtoBGC-823with1ipofe咖1me'5、andthetransfectionefficiencyⅥ娜provedbyFluoresc饥ceMicroscopy.RT.PCRwe‘eusedt0detefmmethemRNAexpressioIlsofGMⅡ、CD44andE。seledmaRertransfection.wresternblot、ⅣereuSedt0detecttheprotemeXpressionsofGMII、CD44andE.selectiIlaRer仃ansfection.AMTTassayandanadhesionteStwereusedt0一⋯一dete嗍inetheB(祀.823Cell’sadh6、esiVeabilityaRer仃aIlSfectiorLResults:TheGMII.shI矾AplasmidwassuCcessfullytmllsfected.Rr-PCRandwest锄blota11alysisindicatedthatmRNAandproteineXpressionofGM11wasdown-regulatedsignifiCantlyi11gastriccancercelllineBGC-823aRertms触ion(P<0.05).MrrandadhesionteStshowedthatthegroupwithGMII。s删Aledtoa10weradh7、esiveabil“ytIlanothers(P<0.05).Comparet0theC0ntrol鲈oup,‘hegrouptransf&tedwithGMII.shI矾Ashowed0bViously10w可CD44n‘武NAanadproteinexpreSsion(P<0.05).AmdE—selectill’smI冰Aandprotein能presslonwaSn0‘changeobViously.Conclusion:T
4、,黏附EFFECToFRNAINTERFERENCETARGETINGGMⅡoNADHESIoNoFGASTRICCELLSABSTRACTObjective:TranSfectedthes删AplasmidofhumanG0培i弘lna彻osldaseII(GMII)t0gastriccancercenIineBGC一823,identi匆itsexpressioniIlBGC-823,andinvestigateitse腩ctonadhesionofthecells·Memods:Trans胁edtheGMII.shRNAplasmidtoBGC-823with1ipofe咖1me'
5、andthetransfectionefficiencyⅥ娜provedbyFluoresc饥ceMicroscopy.RT.PCRwe‘eusedt0detefmmethemRNAexpressioIlsofGMⅡ、CD44andE。seledmaRertransfection.wresternblot、ⅣereuSedt0detecttheprotemeXpressionsofGMII、CD44andE.selectiIlaRer仃ansfection.AMTTassayandanadhesionteStwereusedt0一⋯一dete嗍inetheB(祀.823Cell’sadh
6、esiVeabilityaRer仃aIlSfectiorLResults:TheGMII.shI矾AplasmidwassuCcessfullytmllsfected.Rr-PCRandwest锄blota11alysisindicatedthatmRNAandproteineXpressionofGM11wasdown-regulatedsignifiCantlyi11gastriccancercelllineBGC-823aRertms触ion(P<0.05).MrrandadhesionteStshowedthatthegroupwithGMII。s删Aledtoa10weradh
7、esiveabil“ytIlanothers(P<0.05).Comparet0theC0ntrol鲈oup,‘hegrouptransf&tedwithGMII.shI矾Ashowed0bViously10w可CD44n‘武NAanadproteinexpreSsion(P<0.05).AmdE—selectill’smI冰Aandprotein能presslonwaSn0‘changeobViously.Conclusion:T
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