BMP-2 Up-Regulates PTEN Expression and Induces Apoptosis of Pulmonary Artery Smooth Muscle Cells under Hypoxia 英文参考文献.docVIP

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BMP-2 Up-Regulates PTEN Expression and Induces Apoptosis of Pulmonary Artery Smooth Muscle Cells under Hypoxia 英文参考文献.doc

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BMP-2 Up-Regulates PTEN Expression and Induces Apoptosis of Pulmonary Artery Smooth Muscle Cells under Hypoxia 英文参考文献

BMP-2Up-RegulatesPTENExpressionandInduces ApoptosisofPulmonaryArterySmoothMuscleCells underHypoxia WeifengPi1.,XuejunGuo1*.,LipingSu2,WeiguoXu1* 1DepartmentofRespiratoryMedicine,XinhuaHospital,SchoolofMedicine,JiaotongUniversity,Shanghai,China,2DepartmentofBioengineering,NationalUniversityof Singapore,Singapore,Singapore Abstract Aim:Toinvestigatetheroleofbonemorphogeneticprotein2(BMP-2)inregulationofphosphataseandtensinhomologue deletedonchromosometen(PTEN)andapoptosisofpulmonaryarterysmoothmusclecells(PASMCs)underhypoxia. Methods:NormalhumanPASMCswereculturedingrowthmedium(GM)andtreatedwithBMP-2from5–80ng/mlunder hypoxia(5%CO2+94%N2+1%O2)for72hours.GeneexpressionofPTEN,AKT-1andAKT-2weredeterminedbyquantitative RT-PCR(QRT-PCR).ProteinexpressionlevelsofPTEN,AKTandphosph-AKT(pAKT)weredetermined.ApoptosisofPASMCs weredeterminedbymeasuringactivitiesofcaspases-3,-8and-9.siRNA-smad-4,bpV(HOpic)(PTENinhibitor)andGW9662 (PPARcantagonist)wereusedtodeterminethesignallingpathways. Results:ProliferationofPASMCsshoweddosedependenceofBMP-2,thelowestproliferationratewasachievedat60ng/ml concentration under hypoxia (82.262.8%). BMP-2 increased PTEN gene expression level, while AKT-1 and AKT-2 did not change. Consistently, the PTEN protein expression also showed dose dependence of BMP-2. AKT activity significantly reducedinBMP-2treatedPASMCs.Increasedactivitiesofcaspase-3,-8and-9ofPASMCswerefoundafterculturedwith BMP-2.PTENexpressionremainedunchangedwhenSmad-4expressionwasinhibitedbysiRNA-Smad-4.bpV(HOpic)and GW9662(PPARcinhibitor)inhibitedPTENproteinexpressionandrecoveredPASMCsproliferationrate. Conclusion:BMP-2increasedPTENexpressionunderhypoxiainadosedependentpattern.BMP-2reducedAKTactivityand increased caspase activity of PASMCs under hypoxia. The increased PTEN expression may be mediated through PPARc signallingpathway,insteadofBMP/Smadsignallingpathway. Citation: Pi W, Guo X, Su L, Xu W (2012) BMP-2 Up-Regulates PTEN Expression and Induces Apoptosis of Pulmonary Artery Smoo