SOD1抑制剂LCS-1对弥漫性大B细胞淋巴瘤细胞凋亡的影响

SOD1 Inhibitor LCS-1 Induces Apoptosis in Diffuse Large B-Cell Lymphoma Cells

目的:探讨超氧化物歧化酶1(SOD1)在弥漫性大B细胞淋巴瘤(DLBCL)患者肿瘤组织及DLBCL细胞系中的表达,观察SOD1抑制剂LCS-1对DLBCL细胞系增殖与凋亡的影响,并分析其可能的作用机制。方法:采用免疫组化法检测SOD1在DLBCL组织及反应性淋巴结增生组织中的表达水平,Western blot法检测SOD1蛋白在DLBCL细胞株(TMD-8、OCI-Ly10、OCI-Ly18、OCI-Ly19)中的表达情况。用不同浓度的LCS-1处理DLBCL细胞株后,分别采用CCK-8法检测细胞增殖活力、Western blot法检测SOD1蛋白的表达水平、TUNEL法检测细胞凋亡情况。通过KEGG数据库分析SOD1高表达组的基因富集情况。结果:DLBCL患者肿瘤组织和DLBCL细胞系TMD-8、OCI-Ly18、OCI-Ly19明显高表达SOD1。SOD1抑制剂LCS-1对高表达SOD1蛋白的DLBCL细胞株TMD-8、OCI-Ly18、OCI-Ly19的活力均表现出一定的抑制效应,且呈浓度及时间依赖性(r=0.730,r=0.929,r=0.976);OCI-Ly18、OCI-Ly19细胞株经不同浓度的LCS-1处理后,SOD1蛋白表达水平随LCS-1浓度的增加而下降(r=0.860,r=0.970);LCS-1在3μmol/L浓度时可以显著促进DLBCL细胞株OCI-Ly18、OCI-Ly19的细胞凋亡(P<0.001)。KEGG基因富集通路分析显示,在DLBCL中,SOD1可能通过氧化磷酸化(P=0.002,FDR=0.003)、核糖体(P=0.004,FDR=0.005)等途径发挥作用。结论:DLBCL患者肿瘤组织中SOD1的表达水平明显升高。LCS-1作为SOD1抑制剂,可以明显抑制DLBCL细胞株OCI-Ly18、OCI-Ly19的细胞活力与增殖,促进细胞凋亡,这为DLBCL的治疗提供了新的思路。

Objective:To investigate the expression of superoxide dismutase 1(SOD1)in tumor tissue of patients with diffuse large B-cell lymphoma(DLBCL)and in DLBCL cell lines,to explore the effect of SOD1 inhibitor LCS-1 on proliferation and apoptosis of DLBCL cell lines and analyze its possible mechanisms of action.Methods:Immunohistochemistry assay was used to detect the expression level of SOD1 in DLBCL tissues and reactive lymph node hyperplasia tissues.The expression levels of SOD1 protein in DLBCL cell lines(TMD-8,OCI-Ly10,OCI-Ly18,OCI-Ly19)were detected by Western blot.After the DLBCL cell lines were treated with different concentrations of LCS-1,the cell proliferation activity was detected by CCK-8 assay,the expression levels of SOD1 protein was detected by Western blot,and the cell apoptosis was detected by TUNEL method.The genes enrichment of the SOD1 high expression group were analyzed by the KEGG database.Results:The expression levels of SOD1 in the tumor tissues of DLBCL patients and DLBCL cell lines TMD-8,OCI-Ly18,and OCI-Ly19 were significantly increased.SOD1 inhibitor LCS-1 showed a certain inhibitory effect on the activity of DLBCL cell lines TMD-8,OCI-Ly18,and OCI-Ly19 in a concentration-and time-dependent manner(r=0.730,r=0.929,r=0.976).After being treated with different concentrations of LCS-1,the expression level of SOD1 protein in OCI-Ly18 and OCI-Ly19 cell lines decreased in a concentration-dependent manner(r=0.860,r=0.970);LCS-1 significantly promoted the apoptosis of DLBCL cell lines OCI-Ly18 and OCI-Ly19at a concentration of 3μmol/L(P<0.001).KEGG enrichment analysis suggested that SOD1 may play an important role through oxidative phosphorylation(P=0.002,FDR=0.003)and ribosome(P=0.004,FDR=0.005)pathways in DLBCL.Conclusion:The expression levels of SOD1 in tumor tissues of DLBCL patients were significantly increased.As a SOD1inhibitor,LCS-1 can significantly inhibit the viability and proliferation of DLBCL cell lines OCI-Ly18 and OCI-Ly19,and promote cell apoptosis,which provides a new idea for the treatment of DLBCL.