| Background and Objective: Diffuse large B cell lymphoma(DLBCL)is the most common type of non-Hodgkin's lymphoma,accounting for more than 40% of invasive lymphoma.DLBCL has a high degree of invasiveness and heterogeneity,and its morphology,immunophenotype,genotype and clinical manifestations show great differences.The treatment is mainly targeted therapy with combination chemotherapy and rituximab.Although higher clinical remission rate and survival rate have been achieved,more than 50% of patients with DLBCL still have refractory or relapse after first-line standard treatment.At present,the index of International Prognostic Index(IPI),which is clinically effective in predicting the efficacy or recurrence of patients with diffuse large B-cell lymphoma,is a combination of multiple clinical indicators and can not accurately reflect the heterogeneity of molecular biology.Therefore,it is urgent to improve the prognosis evaluation system of DLBCL patients to guide clinical treatment.In recent years,more and more studies have shown that the occurrence and development of DLBCL are closely related to micro RNAs(miRNAs).Previously,micro RNA-296-5p(miRNA-296-5p)expression in DLBCL was significantly higher than that in lymph node-responsive hyperplasia tissue by using gene chip scanning technique.In addition,RT-q PCR was used to detect the expression of miRNA-296-5p 83 cases of lymph node reactive hyperplastic tissue paraffin specimens confirmed this conclusion.In view of this,we mainly studied the expression of miRNA-296-5p in diffuse B cell lines and its possible role in tumorigenesis and development.Methods:(1)Peripheral blood samples of 50 healthy volunteers were collected,RNA was extracted from mononuclear cells,and the expression of miRNA-296-5p in mononuclear cells and three DLBCL cells was detected by quantitative reverse transcription polymerase chain reaction(RT-q PCR);(2)According to the sequence of miRNA-296-5p,miRNA-296-5p inhibitor and its negative control were synthesized.The synthesized miRNA-296-5p inhibitor and its negative control were transfected into SUL,DB and OCI-LY10.The RT-q PCR was used to detect the expression of miRNA-296-5p to find the most suitable concentration for transfection;(3)SUL and DB were transfected at the most suitable transfection concentration.The proliferation of SUL and DB cells transfected with miRNA-296-5p inhibitor 24,48,and 72 h were detected by CCK-8 assay;(4)Transwell migration assay was used to detect the changes of cell migrationability of DB cells transfected with miRNA-296-5p inhibitor;(5)The apoptosis of DB cells transfected with miRNA-296-5p inhibitor was detected by flow cytometry;(6)The m RNA and protein expression of P53 and PLK1 after transfection were detected by RT-q PCR and Western blot(WB),and it was speculated that miRNA-296-5p might affect the mechanism of tumorigenesis.Results:(1)The expression of miRNA-296-5p in SUL,DB and OCI-LY10 cells was significantly higher than that in peripheral blood mononuclear cells;(2)RT-q PCR results showed that miRNA-296-5p expression was significantly down-regulated in miRNA-296-5p inhibitor group(P<0.01),miRNA-296-5p in miRNA-296-5p inhibitor NC group expression had no significant change(P>0.05);(3)The proliferation of miRNA-296-5p inhibitor group and miRNA-296-5p inhibitor group increased with time after cultured for 24,48 and 72 h in negative control group and interference group compared with control groupand miRNA-296-5p inhibitor NC group was significantly slower(P<0.05),there was no significant difference in OD value between control group and miRNA-296-5p inhibitor NC group;(4)The results of Transwell migration chamber showed that the number of miRNA-296-5p inhibitor cells in the mi R-296-5p inhibitor group was significantly less than that in the control group and the mi R-296-5p inhibitor NC group(P<0.01).There was no significant difference in the number of cells passing through the mi R-296-5p inhibitor NC(P>0.05);(5)The results of flow cytometry showed that there was statistically different in apoptosis between control,mi R-296-5p inhibitor NC and mi R-296-5p inhibitor 3 cells(P<0.05);(6)P53 and PLK1 m RNA and protein did not change significantly before and after transfection.Conclusion:(1)MiRNA-296-5p is highly expressed in diffuse large B-cell lymphoma.(2)Inhibiting the expression of miRNA-296-5p in DLBCL cells can reduce the proliferation and migration of tumor cells,and also cause a slight increase in the apoptosis rate of tumor cells.(3)After inhibiting the expression of miRNA-296-5p in DLBCL cells,there was no significant change in the m RNA and protein expression of TP53 and PLK1,which may not be the direct target genes of miRNA-296-5p. |
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