摘要
目的:通过来那度胺处理过表达TNFAIP3/A20基因后的弥漫性大B细胞淋巴瘤(DLBCL)细胞系OCI-LY3,检测该处理后细胞系的变化及与RNF138、MYD88、p65表达的关系,探讨增强来那度胺治疗DLBCL疗效的可能新靶标。方法:Sanger法测定DLBCL细胞系MYD88突变;CCK-8法测定来那度胺对OCI-LY3细胞增殖活性的影响,筛选细胞IC50;制备TNFAIP3/A20过表达质粒和慢病毒,转染OCI-LY3,得到TNFAIP3/A20基因高表达稳转细胞株OCI-LY3-A20;来那度胺(浓度为367μmol/L)处理OCI-LY3-A20,CCK-8检测用药前后细胞增殖活性的变化,Western blot检测TNFAIP3/A20、p65、RNF138、MYD88蛋白的表达。结果:来那度胺处理OCI-LY3-A20引起细胞内TNFAIP3/A20蛋白表达降低。OCI-LY3-A20中加入来那度胺进一步检测其增殖活性,OCI-LY3-A20较OCI-LY3-MCS(阴性对照病毒组)增殖活性降低;OCI-LY3-A20-Len(即加入来那度胺处理)较OCI-LY3-A20-NC(即未加入来那度胺处理)、OCI-LY3-MCS-Len增殖活性降低。比较OCI-LY3-MCS-NC组、OCI-LY3-MCS-Len组、OCI-LY3-A20-NC组和OCI-LY3-A20-Len组培养细胞蛋白表达水平,其结果表明TNFAIP3/A20、RNF138及MYD88蛋白表达相关,它们可能是影响OCI-LY3细胞增殖活性的重要靶点分子,在临床上具有潜在的治疗价值。结论:NF-κB抑制不是来那度胺发挥作用的主要机制,TNFAIP3/A20可能是来那度胺发挥作用的重要靶点分子,且RNF138及MYD88蛋白参与其中,是潜在的治疗靶点,具有重要临床诊疗价值。
Objective:The diffuse large B-cell lymphoma(DLBCL)cell line OCI-LY3 was treated with lenalidomide after overexpression of the TNFAIP3/A20 gene,and the changes of the cell line and its relationship with the expression of RNF138,MYD88 and p65 were detected,so as to explore the possible new targets to enhance the efficacy of lenalidomide in the treatment of DLBCL.Methods:Sanger method detected the MYD88 mutation of DLBCL cell line.CCK-8 method was used to determine the effect of lenalidomide on the proliferation activity of OCI-LY3 cells,and to screen cell IC 50 and the optimal drug treatment time.By preparing TNFAIP3/A20 overexpression plasmid and lentivirus,OCI-LY3 cells were transfected with them to obtain a highly expressed stable cell line OCI-LY3-A20.Lenalidomide(367μmol/L)treated OCI-LY3-A20 cell line.CCK-8 detected the changes of cell proliferation activity before and after treatment,and Western blot detected the expression changes of NFAIP3/A20,p65,RNF138,MYD88 proteins.Results:The treatment of OCI-LY3-A20 with lenalidomide caused a decrease in the expression of TNFAIP3/A20 protein in the cells.The addition of lenalidomide to OCI-LY3-A20 was further examined for the proliferative activity,and the proliferative activity was decreased by the addition of lenalidomide to OCI-LY3-A20 compared with OCI-LY3-MCS(negative control virus group).OCI-LY3-A20-Len(with lenalidomide)showed reduced proliferative activity compared to OCI-LY3-A20-NC(without lenalidomide)and OCI-LY3-MCS-Len.Comparing the protein expression levels of cultured cells in OCI-LY3-MCS-NC group,OCI-LY3-MCS-Len group,OCI-LY3-A20-NC group and OCI-LY3-A20-Len group,the results showed that TNFAIP3/A20,RNF138 and MYD88 protein expression were related.They may be important target molecules affecting the proliferation activity of OCI-LY3 cells,and have potential therapeutic value in clinical practice.Conclusion:NF-κB inhibition is not the main mechanism for the action of lenalidomide,but TNFAIP3/A20 may be an important target molecule for the action of lenalidomide,and RNF138 and MYD88 proteins are involved in it,which is a potential therapeutic target and has important clinical diagnostic and therapeutic value.
作者
徐丹
徐睿欢
杨文秀
XU Dan;XU Ruihuan;YANG Wenxiu(Department of Pathology,the Affiliated Hospital of Guizhou Medical University,Guizhou Guiyang 550001,China)
出处
《现代肿瘤医学》
CAS
2024年第8期1429-1434,共6页
Journal of Modern Oncology
基金
贵州省科技计划项目(编号:黔科合支撑[2020]4Y147号)
贵州省贵阳市科技计划项目(编号:筑科合同(2019)9-1-6号)。