肽核酸靶向抑制线粒体融合逆转人肺腺癌干细胞顺铂耐药的分析

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1、AbstractandWesternblottechniquerespectivelydetectthemRNAandproteinexpressionofMitofusin1,Mitofusin2,Opa1andAtad3ainA549cellspheresandA549cells.FlowcytometryinstrumentdetectmitochondriamembranepotentialofA549cellsandA549cellspheres;observingthedistributionofmitochondriainA549cellspheresa

2、ndA549cellsbyLaserConfocalMicroscop(LSCM);Solidphasepeptidesynthesispeptidenucleicacid-threebenzenephosphoruscompounds.LSCMobservatethetransfectionandidentificationofpeptidenucleicacid-threebenzenephosphoruscompoundsinlungcancerstemcells.RT-PCRandWesternblotdetectethemtDNAgeneexpression

3、lungcancerstemcellsandwesternblotdetectetheexpressionofmitochondrialfusionrelatedproteinMfn1,Mfn2,Opa1andAtad3aafterthetransfectionandinterferenceofthreebenzenephosphoruscompound;ATPBioluminescenceAssayKitwasusedtoanalyzeATPcontentinA549cellspheresandA549cellafterthetransfectionandinter

4、ferenceofthreebenzenephosphoruscompound;Flowcytometrytestapoptosicrateandmitochondrialmembranepotentialinallgroups.Result:ThemRNAandproteinexpressionofmitochondriafusionrelatedproteinMfn1,Mfn2,Opa1andAtad3ainA549cellspheresincreasedsignificantly;MitochondrialdistributioninA549cellsphere

5、saroundthenuclearandmitochondrialmembranepotentialintheA549cellspheresarehigher;Afterpeptidenucleicacid-threebenzenephosphoruscompoundsactedonmitochondriafusioninA549cellspheresandA549cell,theexpressionofmitochondrialfusionrelatedproteinMfn1,Mfn2,Opa1decreased,whiletheexpressionofAtad3a

6、isnochange,atthesametimetheATPcontentwereallreduced,thecellapoptosisrateincreasedsignificantlyandthemitochondrialmembranepotentialdecreasedobviously.Conclusion:1.ThemRNAandproteinexpressionofmitochondriafusionrelatedproteinMfn1,Mfn2,Opa1andAtad3ainA549cellspheresincreasedsignificantlyth

7、aninA549cells,peptidenucleicacid-threebenzenephosphoruscompoundstargetinginhibitmitochondrialfusionenhancesthesensitivityofcisplatinchemotherapy.2.Peptidenucleicacid-threebenzenephosphoruscompoundsaccumulationinA549lungcancercellspheresastheirhigh∆ψm,closedgeneexpressionofmtDNA