Wnt经典通路β-catenin过表达对人牙周膜干细胞增殖能力及成骨能力的.pdf

Wnt经典通路β-catenin过表达对人牙周膜干细胞增殖能力

及成骨能力的影响

李影;刘娜;张维;顾斌

【摘要】Objective:ToconstructWnt/β-cateningeneoverexpressedvector

andtoinvestigatetheeffectsofitsoverexpressiononproliferationand

osteogenicabilityofhumanperiodontalligamentstemcells.Methods:8

healthypremolarsandthirdmolarsextractedfortreatmentwereisolated

andculturedforhumanhealthyindividuals(PDLSCs).AmplifytheWnt/β-

cateningenefromPDLSCs,andcloneditintopLV-EGFP-Nvector,to

constructWnt/β-cateninoverexpressedvector,thentotransfectitinto

PDLSCsbylipofectionmethod.Cellmorphologywasobservedunder

fluorescentmicroscopeaftertransfectionofPDLSCsinconventionaland

osteogeniccultureconditionontheseventhday.WetestedMTTresultsof

PDLSCs.lymphoidenhancerfactor-1(LEF1),Tcellfactor-4(LEF4),Runt-

relatedtransferfactor-2(Runx2),alkalinephosphatase(ALP),humantypeI

collagen(COL-1)andP38mitogenactivatedproteinkinase(P38MAPK)

weredetectedbyreal-timequantitativePCR.Results:GFPwasexpressed

efficientlyinPDLSCsaftertransfection.MTTresultsshowedproliferationof

PDLSCsaftertransfectionwasboostedspecificallyandefficientlythan

before.Real-timePCRresultsshowedtheexpressionofLEF1mRNAwas

significantlyincreasedthanbefore,whilethereisnodifferencein

TCF4.Simultaneously,Runx2,ALP,COL-1andp38weresignificantly

decreasedofPDLSCstransfectedwithWnt/β-cateninoverexpressedvector

culturedinosteogenicdifferentiationcondition.Conclusion:ClassicWnt/β-

cateninpathwayplaysanimportantroleinPDLSCsproliferationand

osteogenicdifferentiation.OverexpressionofthecoreproteinWnt/β-

catenincanupregulatetheexpressionofLEF1,promotetheproliferationof

PDLSCs,andinhibittheosteogenicabilityofPDLSCsbydownregulating

theexpressionofRUNX2,ALP,COL-1andP38.%目的:探讨Wnt经典通路β-连

环蛋白(β-catenin)过表达对人牙周膜干细胞(periodontalligamentstem

cel