| Background:Diffuse large B cell lymphoma（DLBCL）is the most common type of non-Hodgkin’s lymphoma and is characterized as a high degree of heterogeneity from cell of origin to clinical course and prognosis.Recent years,RIP2 has been proven to play a key role in the inflammatory process as well as affecting the development of neoplastic diseases,including hematological malignancies such as lymphomas.What’s more,the dysfunction of the PI3K/Akt pathway is often closely related to the downstream molecular mutations in DLBCL.Objective:To investigate the expression and prognostic significance of RIP2 in diffuse large B-cell lymphoma,and to explore the role of RIP2 in the life process of DLBCL cells and to search for potential therapeutic targets through cell functional experiments.Methods:This study mainly consists of two parts:（1）To investigate the expression of RIP2 in GCB and non-GCB DLBCL verified by immunohistochemical staining,and to detect the expression of bcl-2 and c-myc in combination with related clinicopathological datas to analyze the prognostic significance of RIP2 in DLBCL;（2）To explore the effect of RIP2 on the life process of DLBCL by cell functional experiments and to the relevant mechanisms of its function.In this study,we selected GSK583,a kind of chemical synthesis,and the specific small interfering RNA（RIP2-siRNA）to inhibit the expression of RIP2 in DLBCL cells.（1）The cell viability was detected by CCK8 assay,and the effect of RIP2 silencing on the proliferation of DLBCL cells was observed;（2）The effect of RIP2 inhibition on the apoptosis of DLBCL cells was detected by Annex V-FITC staining,Caspase-3 activity assay and bcl-2/c-myc.（3）After treated with GSK583 and transfected with RIP2-siRNA,the expression of p65 and PI3K/AKT protein in DB and oci-ly10 cells were detected by western blot in order to elucidate the specific mechanism of RIP2-related DLBCL cell growth inhibition.Results:（1）The relationship between RIP2+/-and clinicopathological parameters:（1）The expression of RIP2 was associated with IPI scores attributed to middle and high risk group and the An arbor stage ofⅢ,Ⅳ（p<0.05）.（2）The survival rate of DLBCL in GCB was higher than that in non-GCB（p=0.028）.The survival rate of DLBCL in RIP2-positive group was lower than that of RIP2-negative group（p=0.013）.The patients with RIP2-positive DLBCL received R-CHOP presented a better survival rate than that received CHOP（p=0.001）.（3）The expression of RIP2 in DLBCL cell line was higher than that in peripheral blood mononuclear cell from healthy people（p=0.001）,and it was differentially expressed in different subtypes（p<0.001）.（2）In DB and oci-ly10 two DLBCL cells:（1）Both GSK583 and RIP2-siRNA transfected cells could inhibit cell viability and promote cell apoptosis rate;（2）In DB,the GCB-type of DLBCL cells,inhibition of RIP2 leading to bcl-2/bax-dependent cell apoptosis was associated with a decrease of p65 expression,whereas in the ABC-type of DLBCL cells oci-ly10 was independent of bcl-2/bax.（3）Treatment with GSK583 in DB and oci-ly10reduced the phosphorylation level of RIP2 and Akt in DLBCL cells.Conclusion:（1）The expression of RIP2 may be associated with poor prognosis of DLBCL and specific subtypes,such as bcl-2/c-myc double-expression lymphoma.（2）Inhibition of RIP2 expression can significantly affect the growth of DLBCL cells,it may participate the cell apopotosis of GCB-type of DLBCL cells through the inhibition of p65 expression.As a RIP2 inhibitor,GSK583 can inhibit the life process of DLBCL cells by negatively regulating PI3K/AKT pathway,which may become a new target of DLBCL therapy. |
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