| Objective:To investigate the expression of eukaryotic initiation factor 3b(eIF3b)in Chronic myeloid leukemia(CML)and its role in the development and progression of CML.Methods:Collected 50 cases bone marrow biopsy pathological specimens of patients diagnosed chronic myelogenous leukemia chronic phase(CP-CML)from February2016 to November 2018 in our and other hospital.and collected 50 cases of Orthopedic trauma patient from our department during the same period.All cases were divided into experimental group and control group.And specimens were detected of eukaryotic initiation 3b(eIF3b),G protein coupled receptor antibody releasing factor 2(GRF2),cycle dependent kinase inhibitors(P27)expression of polyclonal antibody monoclonal,Analysis of the experimental group and control group in the index to express quantities.The correlation between BCR-ABL1 copy number and eIF3b expression level was analyzed,and the correlation between the time to obtain complete cytogenetic response and eIF3b expression level was analyzed.COX analysis was performed on age,gender,BCR/ABL copy number,eIF3b and complete cytogenetic response and major molecular biological response at the initial diagnosis in the experimental group.CML cell lines TK6 and K562 were selected as subjects to investigate the effect of eIF3b on CML cell function,including qPCR(real-time fluorescence quantitative nucleic acid amplification fluorescence detection system)to detect the expression of eIF3b in CML cell line,and to construct eIF3b lentivirus interference vector sheIF3b and its control shCtrl.Cell lines were transfected,Western Blot was used to detect eIF3b protein expression,qrt-pcr was used to detect interference efficiency,CCK8 was used to detect eIF3b cell proliferation,flow detection was used to detect eIF3b cell cycle and apoptosis.Results:1.EIF3B positive brown-yellow granules were expressed on CML cytoplasm and cell membrane.In the mature granulocyte of EIF3B positive brown-yellow granules in the control bone marrow samples,the positive expression rate of EIF3B in bone marrow tissue of CML patients was 76.9%.The positive rate of expression was 38.5%(?~2=14.729,P=0.000),the mean value of CML group±standard deviation(~_X±S):0.031±0.027;the control group(~_X±S)was 0.025±0.010;the expression of the two groups was significantly different(F=5.585,P=0.017);GRF2-positive brown-yellow granules were expressed in CML cytoplasm in both experimental and control groups.The positive rate of expression in bone marrow tissue of CML patients was 75.6%,and the positive rate of control group was 68.5%(?~2=0.407,P=0.523).Mean value of standard deviation of standard CML group(~_X±S):0.031±0.027;control group(X~_±S)was(0.025±0.010);there was no significant difference in expression between the two groups(F=2.155,P=0.145).P27 positive brown-yellow granules were expressed in CML cytoplasm,The positive rate of expression in bone marrow tissue of CML patients was 69.6%,and the positive rate of control group was 63.5%(?~2=0.794,P=0.373),and standard deviation in CML group(~_X±S):0.012±0.012;control group(X~_±S)was0.010±0.008;poor expression in both groups The difference was not statistically significant(F=0.652,P=0.421).2.There was a positive correlation between BCR-ABL1 copy number and EIF3B expression in the CML experimental group(r=0.514,P=0.000);there was a positive correlation between the number of complete cytogenetic remission days and EIF3B expression(r=0.393,P=0.005);there was no significant difference between the time of major molecular biological remission and the expression of EIF3B(r=0.038,P=0.751).3.Multiple regression COX analysis of CML patients with age,gender and EIF3B expression,no correlation between BCR/ABL copy number at the time of initial diagnosis,complete cytogenetic remission time EIF3B expression was correlated,the main molecular biological remission time and EIF3B expression Correlation.4.qPCR detection of mRNA levels eIF3b was highly expressed in TK-6,K562and Jurkat,and was highly expressed in the former two.5.The results of CCK8 assay showed that there was a statistically significant difference in cell proliferation between the sheIF3b group and the sheIF3b group in the TK6 cell line.The knockdown rate at 24 h(t=18.933,P=0.000);the 48h knockdown rate(t=7.811,P)=0.000);72h knockdown rate(t=55.618,P=0.000).In K562 cell line,the proliferation rate of sheIF3b group at 24h compared with shCtrl group(t=3.544,P=0.009);48h increase rate(t=16.747,P=0.000);72h proliferation rate(t=15.985,P=0.000).There was a statistically significant difference in cell proliferation between the two groups in the sheIF3b group.6.eIF3b interferes with the proliferation cycle of CML cells,which increases the number of cells in S and G1 phases,and reduces the number of cells in G2/M phase.Conclusion:1.EIF3B is highly expressed in CML,and the expression intensity is related to cytogenetics,suggesting that this factor is involved in the development of CML2.Cell function experiments confirmed that interference of the target gene eIF3b inhibited the proliferation and promoted apoptosis of chronic myelogenous leukemia cell lines tk-6 and K562. |
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