The Role Of MicroRNA-106a In Lymphoma Pathogenesis

Objective: Malignant lymphoma (ML)is the primary in the lymph nodesand (or) junction of lymphoid tissue malignant tumor,it is one of the mostcommon malignant tumor in children.At present its pathogenesis is not fullyclear,this have cause confusion to clinical diagnosis and treatment,so it havebecom a serious threat to children’s health.Therefore dedication to thecause,early detection,dynamic view,all need a simple easy high specificitymarker as clinical indicator,these are very important to improve the cure rateand survival rate of children’s lymphoma.Many studys showed thatmiRNA-106a closely related to normal growth,development and tumorige-nesis.In a variety of human tumor,miRNA-106a has been showed up-regulatedexpression,so considered it to be a carcinogenic miRNAs.The reports aboutthe expression of miRNA-106a in malignant lymphoma is less,and is how toplay a role in lymphoma cells control has not yet been reported.ThepRb/E2F1cell regulation pathway is closely related to tumrigenesis,There areRb1(retinoblastoma gene), E2F1(transcription factor)and so on in the pathway,recent research found that microRNA have appeared in the pathway,In thispathway,when the synthesis of Rb1is reduced,the activity of E2F1is notmonitored,will cause the disorder of cell cycle and unlimited proliferation,make the activation of Caspase-3reduced,then the tumor cell proliferation.Rb-1is one of the anti-oncogene,is one of downstream target genes ofmiRNA106-a, miR-106a by combining with Rb1mRNA,inhibit the translationof Rb1mRNA,lead to decrease the production of Rbl protein,make the cellproliferation not be controlled;E2F1is a member of E2F transcription factorfamily,its expression depends on Rb1,it is a very important regulatory factorfor cell cycle to across G1phase to S phase,it plays a key role in the regulationof cell cycle process and cell proliferation;Caspase-3it is the core factor of theregulation of apoptosis in cell,it is the most trigger apoptosis,finally all is mediated by Caspase-3give play to the role of apoptosis signal transductionpathway.Method: Culture human T cells lymphoma Jurkat in vitro asexperimental group1,various concentration of adriamycin treat Jurkat asexperimental group2,set up20cases of healthy human peripheral bloodmononuclear cells as control group.miR-106a was measured using Real TimePCR,Rb1.E2F1.Caspase-3were measured using Reverse Transcription-Polymerase Chain Reaction,inhibition of the proliferation intervened byAdriamycin was measured using MTT assay,Flow cytometry (FCM) analysisof the cell cycle changes of Jurkat after intervened by Adriamycin,as well asthe detection of apoptosis.to known the effects of Adriamycin on the abovefactors.Using SPSS13.0to analyze the data, P<0.05has significant difference.Results：1MTT results display:Compared with the experimental group1,theabsorbance value (OD) in experimental group2after intervened by variousconcentration of Adriamycin for24h,48h,72h,all have varying degrees ofdecline.Except for intervened by various concentration of Adriamycin for24hcompared with the experimental group1no statistical difference (P>0.05),intervened by various concentration of Adriamycina for48,72h compared withexperiment1group were statistically significant (P<0.05).And the absorbancevalue among various concentration groups and various time has statisticaldifference (P<0.05).2Real Time PCR results display:In the experimental group1,the expression ofmiR-106a is significantly higher than the control group,with statisticalsignificance (P<0.05).In experiment group2,after intervente by variousconcentration of Adriamycina for48,72h,the expression of miR-106a are allsignificantly lower than experimental group1,and with the concentrationincreasing, time extending,the expression decling,all with statisticalsignificance (P<0.05).3RT-PCR results display:In the experimental group1,the expression ofRb1,Caspase-3are all significantly lower than the control group,at the same time,the expression of E2F1are all significantly higher than the controlgroup,all with statistical significance (P<0.05).In experiment group2,afterintervente by various concentration of Adriamycina for48,72h,the expressionof Rb1,Caspase-3are all significantly higher than the experimental group1,all with the concentration increasing, time extending,the expression rising,allwith statistical significance (P<0.05);at the same time,the expression of E2F1are all significantly lower than the experimental group1,all with theconcentration increasing,time extending,the expression declining,all withstatistical significance (P<0.05).4FCM result display:In the experimental group2,after intervented by variousconcentration of Adriamycina for48,72h,the change of the cell cycledistribution are obviously,the G1/S phase cells are increased,G0,G2/M phasecells are reduced,Compared with the control group,the change of G1/S phasecells with statistical significance (P<0.05);at the same time,all appear obviousapoptosis peak,but in the control group,there not appear apoptosis peak,andcompared with the control group,the apoptosis rate all all with statisticalsignificance (P<0.05).5The correlation among miRNA-106a and above three factors afterintervened by various concentration of Adriamycina:In experiment group2,after intervente by various concentration of Adriamycina for48,72h,miR-106a and Rb1,Caspase-3are all positively correlated,miR-106a andE2F1are all negative correlated, all with statistically difference (P<0.01).Conclusion：1After Jurkat intervened by various concentration of Adriamycin,theexpression of miR-106a, E2F1decreased, at the same time,expression of Rb1,Caspase-3increased, the G1/S phase cell number increased,G2/M,G0phasecell number decreased significantly,the apoptosis rate increased,suggest cellblock in the G1/S phase may be associated with expressive suppression ofE2F1,lead to the apoptosis of Jurkat.And suggested that miR-106a mayinhibited by Adriamycin.2miR-106a and Rb1,Caspase-3are all positively correlated,miR-106a and E2F1are all negative correlated,suggested that miR-106a may have somerelevance to the pRb/E2F1cell regulation pathway,The reduce of miR-106amade by Adriamycin lead to the pathways changed,thus inhibite theproliferation of Jurkat.3Compared with Control group,the expression of miR-106a in Jurkat cells issignificantly higher,after intervened by with various concentration ofAdriamycin,the expression is significantly decreased,suggested that miR-106amay be by down-regulation the expression of Rb1,make its can’t suppress theproliferation induced by E2F1,then Caspase3activation decreases,it may bethe miR-106a as oncogen play a role of one of the important mechanism byRb/E2F1cell regulation pathway.4Compared with health people,the expression of miR-106a in Jurkat cells issignificantly higher,after intervened by with various concentration ofAdriamycin,the expression is significantly decreased,suggested that miR-106amay as a markers for the diagnosis,treatment,observation of curative effect andtargeted therapy target in lymphoma.