gsdma3基因在小鼠皮肤上皮细胞中对wnt5a ecadherin nfκb表达影响的研究-effects of gsd ma 3 gene on wnt5a e cadherin nf κ b expression in mouse skin epithelial cells.docx

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gsdma3基因在小鼠皮肤上皮细胞中对wnt5a ecadherin nfκb表达影响的研究-effects of gsd ma 3 gene on wnt5a e cadherin nf κ b expression in mouse skin epithelial cells.docx

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gsdma3基因在小鼠皮肤上皮细胞中对wnt5a ecadherin nfκb表达影响的研究-effects of gsd ma 3 gene on wnt5a e cadherin nf κ b expression in mouse skin epithelial cells

缩略语表英文缩写B6英文全称C57BL/6Jmouse中文全称C57BL/6J小鼠Cy3cyclopentadienylanion33H-吲哚菁类染料DPdermalpapilla真皮乳头EDTAethylenediaminetetraacetinacid乙二胺四乙酸EBethidiumbromide溴化乙锭Gsdma3GasderminA3GasderminA3基因/蛋白GFPgreenfluorescenceprotein绿色荧光蛋白ICCimmunocytochemistry免疫细胞化学IHCimmunohistochemistry免疫组织化学mRNAmessengerribonucleicacid信使RNAN1N1expressionplasmid空表达质粒ORSouterrootsheath外根鞘PBSphosphatebufferedsolution磷酸盐缓冲液IRSinnerrootsheath内根鞘RT-PCRreversetranscriptionPCR反转录聚合酶链反应WBwesternblot蛋白免疫印记1GasderminA3geneaffectingtheexpressionofWnt5a,E-cadherinandNF-κBinthemouseskinkeratinocytesAbstractBackground,Gsdma3geneisamemberofGsdmgenefamily,whichbelongstotheGsdmagenesub-family,locatedatthemousechromosome11,closelylinkedtotheretinoicacidreceptorαandkeratin1genes.SixmutantsofGsdma3canbederivedfromtheGsdma3genehotspots,whichcouldresultinirregularhairfolliclecycling,skinhyperplasiaandskininflammation.Ontheotherhand,canonicalWnt／β-cateninpathwayplaythecriticalroleinmasteringtheproliferationanddifferentiationofmouseskinkeratinocyte.SinceourpreviousinvestigationindicatedthatGsdma3mightinfluencethecanonicalWnt／β-cateninpathway,followingobjectiveshavebeenestablished:Objective,1)ToascertaintheeffectofGsdma3onthecanonicalWnt／β-cateninpathwayinhibitorWnt5ainthemouseskinkeratinocyte.ToascertaintheeffectofGsdma3onthecanonicalWnt／β-cateninpathwayinhibitorE-cadherininthemouseskinkeratinocyte.ToascertaintheeffectofGsdma3ontheNF-κBinthemouseskinkeratinocyte.NF-κBisaco-activatorofβ-catenininthehairfolliclemorphogenesisMethods,1)Plasmidtransfection,tissueslide,IHC,RT-PCR,WBwereexploitedtovalidatewhethertheassociationscanbeestablishedbetweenGsdma3andWnt5a/E-cadherin/NF-κBinvivo.2)Cellculture,plasmidtransfection,ICC,RT-PCR,WBwereexploitedtovalidatewhethertheassociationscanbeestablishedbetweenGsdma3andWnt5a/E-cadherin/NF-κBinvitro.Results,1)Invivo,IHCassaysshowedthat,intheskinsofGsdma3mutantmice,strongerpositivesignalsofWnt5aweredetected.GenerescueassaysrepresentedthatweakerpositivesignalsofWnt5aweredetectedintheGsdma3ov