白血病细胞tgf-β受体表达分析-analysis of tgf - β receptor expression in leukemia cells.docxVIP

notbeenreportedwhetheritwasdifferentonTGF-β1bindingwiththetwoisformsofTGF-βRⅡ.NeithertheexpressionofTGF-βRⅠandthetwoisformsofTGF-βRⅡinleukemiacellshasbeenreported.Inthisstudy,TGF-βRⅡanditsisoformsweredetectedbyabsolutequantificationRT-PCR,theexpressionandthecodingsequenceofTGF-βRIinacuteleukemiacellswerealsodetectedbyRT-PCRandDNAsequencinginleukemiacells.AndtheaffinityofTGF-β1withTGF-βRⅡAandTGF-βRⅡBinK562cellswasdetectedbyradiationbindingassay.Further,5casesofacuteleukemiaprimarycells,whichhavedifferentexpressionlevelofTGF-βRIandTGF-βRⅡ,wereculturedtoobserveinhibitationbyaddingdifferentdosesofTGF-β1.Results:TheexpressionofTGF-βRⅡAandTGF-βRⅡBmRNAwerelowerinacuteleukemiacellsthanthatinnormalbonemarrowmononuclearcells(P=0.0057,0.0006);theexpressionofTGF-βRⅡBmRNAwerelowerinchronicleukemiacellsthanthatinnormalbonemarrowmononuclearcells(P=0.0002),buttheexpressionofTGF-βRⅡAmRNAwasnotsignificantlydifferentbetweenchronicleukemiacellsandnormalbonemarrowmononuclearcells(P=0.1215).RT-PCRshowedthattheexpressionlevelofTGF-βRImRNAinacuteleukemiacellsandnormalbonemarrowmononuclearcellsdependonindividuals,therewasnosignificantdifferencebetweenthetwogroups.NodeletionandmutationwereobservedinTGF-βRImRNAcodingsequence.TherewasnosignificantdifferenceontheaffinityofTGF-β1（P=0.8876）betweenTGF-βRⅡAK562cellsandTGF-βRⅡB-K562cells.1ng/mlTGF-β1couldnotinhibittheproliferationof5casesacuteleukemiaprimarycells（P0.05）;5ng/mlTGF-β1couldinhibittheproliferationof3casesacuteleukemiaprimarycellswhichhashigherexpressionofTGF-βRIandTGF-βRⅡ（P0.05）,butthesamedoseTGF-β1couldnotinhibitthe2casesacuteleukemiaprimarycellswhichhaslowerexpressionofTGF-βRIandTGF-βRⅡ（P0.05）;25ng/mland50ng/mlTGF-β1couldinhibittheproliferationofallthe5casesacuteleukemiaprimarycells（P0.05）.Conclusion:TheexpressionofTGF-βRⅡAandTGF-βRⅡBmRNAinacuteleukemiacellswerelowerthanthatinnormalbonemarrowmononuclearcells;however,inchronicleukemiacells,onlyTGF-βRⅡBmRNAwerelowerthanthatinnormalbonemarrowmononuclearcells.Therewasnosignificantdifferen