Effects of Artesunate on DNMT1 expression in highrisk MDS cell strain of SKM-1
WANG Ying, NIU Zhiyun, XING Lina, QIAO Shukai, PAN Ling*
2016, 37(12):
1374-1378.
doi:10.3969/j.issn.10073205.2016.12.003
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[
Abstract ] Objective Toobservetheeffectsofartesunate ( ART ) ongeneexpressionlevels
ofDNMT1byusingSKM-1cellsastheresearchobject , andprovidelaboratoryevidenceinvitro
fortheusageofARTinMDStreatment.Methods AfterbeingtreatedwithART ( 1 , 5and10
μ mol
/ L ), DNMT1mRNAchangesinSKM-1cellswereobservedwithRT-PCR , andtheprotein
expressionchangeswereobservedwith WestBlot.Then , theexpressionchangesofp15INK4b ,
theDNMT1targetinggenewereobservedbyusingreal-timequantitativeRT-PCR , andthe
changesofp15INK4bpromoter methylationstate wereobservedbyusing methylationPCR.
Results AfterbeingtreatedwithART ( 1 , 5and10 μ mol
/ L ), thedifferencewasstatistically
significantforDNMT1geneexpressioninSKM-1cellscomparedwithcontrolgroup ( P <0.05 ),
buttherewasnosignificantdifferenceinthethreedosegroups ( 1 , 5and10 μ mol
/ L )(
P >0.05 ) AstoDNMT1proteinexpression , comparedwith5 μ mol
/ Lgroup , therewassignificantlylower
intheothertwogroups ( 1and10 μ mol
/ L )(
P <0.05 ), andalsosignificantlylowerthanthatin
controlgroup ( P <0.05 ) .Howevertherewasnosignificantdifferenceamongtheothergroups
(
P >0.05 ) .AfterSKM-1cellsbeingtreatedwithARTfor3h , 6hand12h , comparedwith
controlgroup , thedifferenceofDNMT1geneandproteinexpressionsinthethreegroups (
3h , 6
hand12h ) wassignificant ( P <0.05 ) .Besides , thedifferenceofgeneexpressionbetweenthe
twogroups ( 3h , 6h ) andthe12hgroupwasalsosignificant ( P <0.05 ) .Buttherewasno
significantdifferentofproteinexpressioninothertreatedgroups ( P >0.05 ) .AfterSKM-1cells
beingtreatedwithART , p15INK4BmRNAlevelofSKM-1cellsinthethreegroups ( 5and10
μ mol
/ Lgroups , anddecitabinegroup ) wasstatisticallyhigherthanthatincontrolgroup (
P <
0.05 ), withthelevelin10 μ mol
/ Lgroupthehighestinthefivegroups , andthesignificant
differencewasonlyfoundbetween10 μ mol
/ Lgroupandthefourgroups (
controlgroup , 1 μ mol
group , 5 μ molgroupand decitabinegroup )( P <0.05 ) .Afterbeingtreated with ART ,
p15INK4bpromotermethylationlevelwasdecreasedinSKM-1cells , butthedifferencewasnot
significant ( P >0.05 ) .Onthecontrary , thelevelofpromoterunmethylationstateinARTgroup
wasincreasedcomparedwithtwoothergroups ( controlgroupandthedecitabinegroup )( P <
0.05 ) .Conclusion ART can restorethe expression of p15INK4b byinhibit