Expression and mechanism of miRNA-106a in human lymphoma Jurkat cells
Objective To study the expression and mechanism of miRNA-106a in human lymphoma Jurkat cells.Method Take human lymphoma Jurkat cells in logarithmic growth phase,and prepare 5 ml of physiological saline in the culture medium with concentrations of 0,0.5,1.0,and 1.5 μg/ml of doxorubicin.Take blood mononuclear cells from healthy individuals(control group).Quantitative reverse transcription-polymerase chain reaction(qRT-PCR)was used to detect the expression levels of miRNA-106a and retinoblastoma 1(RB1),E2F transcription factor 1(E2F1),and caspase 3 mRNA.Methyl thiazolyl terazolium(MTT)assay was used to detect cell proliferation ability,flow cytometry was used to detect cell apoptosis ability,and Western blot was used to detect the expression levels of RB1,E2F1,and caspase 3 protein.Result The expression levels of miRNA-106a in Jurkat cells of lymphoma treated with 0 μg/ml doxorubicin were higher than that in blood mononuclear cells from control group(P＜0.01).As the concentration of doxorubicin in-creased and the duration of action prolonged,the expression levels of miRNA-106a gradually decreased,the optical den-sity(OD)gradually decreased,and the inhibition rate(IR)of cell proliferation and apoptosis gradually increased,the ex-pression levels of RB1,caspase 3 mRNA and their proteins also gradually increased,while the expression levels of E2F1 mRNA and its protein gradually decreased,and the differences were statistically significant(P＜0.05).The correlation analysis results showed that miRNA-106a were negatively correlated with the expression of RB1 and caspase 3(P＜0.01),and positively correlated with the expression of E2F1(P＜0.01).Conclusion miRNA-106a is highly expressed in human lymphoma Jurkat cells,and may regulate lymphoma progression by regulating the expression of RB/E2F1 path-way related proteins.
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