The Mechanism Of MiR-194 Inhibiting The Proliferation Of Mantle Cell Lymphoma

Objective:This study was aimed to investigate the expression of KDM5 B and miR-194 in Mantle Cell Lymphoma(MCL),in order to reveal the functions of KDM5 B and miR-194 and the possible mechanism in the occurrence and development of MCL.Method:1.Case group: experimental group: 45 cases of mantle cell lymphoma(MCL)with complete clinical data confirmed by histopathology and immunohistochemistry;Control group: reactive proliferative lymphadenitis in 30 cases.2.MCL cell lines: five cell lines(Mino,Z-138,Je Ko-1,RF-1 and REC-1)were selected for in vitro experiments.Mino cells and Je Ko-1 cells were selected to construct the following transgenic cells:(1)With silencing adenovirus of KDM5 B sh RNA and overexpressing adenovirus of KDM5B-GFP.(2)With silencing adenovirus of miR-194 and overexpressing adenovirus of miR-194 mimic.(3)Co-infected with adenovirus miR-194 mimic plus KDM5B-GFP.(4)Peripheral blood mononuclear cells from 5 healthy volunteers were selected as control.3.The expression of KDM5 B protein from patient tissues was detected by immunohistochemistry.4.The cell proliferation with adenovirus infection was checked by MTT.5.The apoptosis after adenovirus infection was detected by flow cytometry.6.The expressions of KDM5 B m RNA and miR-194 in adenovirus infected cells and patient tissues were detected by fluorescence quantitative PCR.7.KDM5 B protein expression in adenovirus infected cells was detected by Western Blot.8.Double luciferase reporting assay was used to comform whether KDM5 B was the target gene of miR-194.9.The cell proliferation was detected by KDM5 B rescue assay.10.Statistical analysis: SPSS 19.0 software was applied for analysis,and homogeneity test of variance and normality test were performed routinely.The data were presented as mean ± standard deviation(mean±SD).T test was used for the comparison between two groups of univariate data,and one-way analysis of variance was used for the comparison between multiple groups of data.Non-parameter was adopted for variance unevenness;Spearman analysis method was used for correlation analysis between the two groups.A value of P<0.05 was defined as statistically significant.Results:1.The expression of KDM5 B protein in control group(16.67%)was lower than that in MCL(48.89%).And the expression of miR-194 in control group(0.991±0.245)was higher than that in MCL(0.223±0.149).The difference was statistically significant(P<0.05).Spearman test showed KDM5 B was significantly negative correlated with miR-194(P<0.05).The result was similar to the test of 5 MCL cell lines.There was not correlation between the expression of KDM5 B or miR-194 and the MCL patient’s age,gender,or symptom of lymphoma group B(P>0.05).But the higher serum β2-MG,the higher the IPI score,the later Ann-Arbor clinical stage.The higher expression of KDM5 B,the lower expression of miR-194,whose difference was statistically significant(P<0.05).2.KDM5 B sh RNA reduced the expression of m RNA and protein of KDM5 B in Mino cell and Je Ko-1 cell,inhibited cell proliferation,and induced apoptosis.The difference was statistically significant compared with the control group(P<0.05).KDM5B-GFP increased the expression of m RNA and protein of KDM5 B in Mino cell and Je Ko-1 cell.Promoted cell proliferation and inhibited apoptosis in Mino cell(P<0.05).But there was no significant change on cell proliferation and apoptosis in Je Ko-1(P>0.05).3.Mi R-94 mimic infected the Mino cell and Jeko-1 cell and reduced the expression of KDM5 B protein in them,inhibiting cell proliferation and inducing apoptosis(P<0.05).Mi R-194 inhibitor infected the Mino cell and Jekou-1 cell,increasing the expression of m RNA and protein of KDM5 B in them,promoting cell proliferation and inhibiting apoptosis in Mino cell(P<0.05),but there was no significant change on cell proliferation and apoptosis in Jeko-1(P>0.05).4.Dual-Luciferase reporter showed that KDM5 B may be one of the miR-194 target genes in MCL.Over-expression LV-KDM5 B partially cancelled effect of miR-194 mimic on inhibiting proliferation in Mino cell and Jeko-1 cell.Conclusions:1.There was a negative correlation between expression of KDM5 B and miR-194 in MCL.Their expressions were closely related to the serum β2-MG,the IPI score and the Ann-Arbor clinical stage.2.Reducing the expression of KDM5 B could inhibit cell proliferation and promote apoptosis,and increasing its expression could promote cell proliferation and inhibit apoptosis.KDM5 B may functions as a oncogene.3.Increasing the expression of miR-194 could inhibit the proliferation,and reducing the expression could promote the proliferation,and may functions as an antioncogene.4.KDM5 B may play as one of the miR-194 target gene which miR-194 could reduce the expression of KDM5 B and inhibit the cell proliferation in MCL.