Objective: To investigate the expression of miR-126 in diffuse large B-cell lymphoma (DLBCL) tissues and its biological function.
Methods: The lymphoma tissues of 46 DLBCL patients in our hospital were selected as the research object, and the lymph node hyperplasia tissue of 31 patients with reactive hyperplasia were selected as controls. The expression level of miR-126 in the patients' tissues was detected by real-time fluorescent quantitative PCR (RT-qPCR), and the correlation of miR-126 expression with the pathological characteristics and prognosis of the patients was analyzed. The DLBCL cell line SU-DHL-4 was transfected with miR-126 inhibitor and its negative control (NC inhibitor) or miR-126 mimics and its negative control (NC mimics). RT-qPCR assay was used to detect the expression level of miR-126 in cells; MTT method was used to detect cell proliferation activity; single clone formation test was used to detect cells colony-forming ability; Annexin V/PI double staining assay was used to detect cell apoptosis; Transwell test was used to detect cell migration and invasion ability; the expression levels of apoptosis-related proteins cleaved-Caspase-3, Bcl-2 and Bax were detected by Western blot.
Results: miR-126 was highly expressed in lymphoma tissues of DLBCL patients, and its expression level was significantly correlated with Hans type, IPI score and Ann-Arbor stage of DLBCL patients (P<0.05). Kaplan-Meier survival analysis showed that the survival rate of DLBCL patients with high expression of miR-126 was significantly lower than that of patients with low expression (P<0.05). Compared with the NC mimics group, the miR-126 expression level, cell proliferation rate, number of colony-forming units, migration and invasion ability, and Bcl-2 protein expression level in the miR-126 mimics group were significantly increased (P<0.05), but the cells apoptotic rate, cleaved-Caspase-3 and Bax protein expression levels were significantly reduced (P<0.05). Compared with the NC inhibitor group, the miR-126 expression level, cell proliferation rate, number of colony-forming units, migration and invasion ability, and Bcl-2 protein expression level in the miR-126 inhibitor group were significantly reduced (P<0.05), but the cells apoptosis rate, cleaved-Caspase-3 and Bax protein expression levels were significantly increased (P<0.05).
Conclusion: miR-126 is highly expressed in lymphoma tissues of DLBCL patients and its expression level is related to the poor prognosis of patients. miR-126 can promote DLBCL cell proliferation, invasion and migration, and inhibit cell apoptosis.
题目: MiR-126在弥漫性大B细胞淋巴瘤组织中的表达及其生物学功能研究.
目的: 探讨miR-126在弥漫性大B细胞淋巴瘤（DLBCL）组织中的表达及其生物学功能.
方法: 选择本院收治的46例DLBCL患者淋巴瘤组织作为研究对象，另选取31例淋巴结反应性增生患者淋巴结增生组织为对照。采用实时荧光定量PCR（RT-qPCR）测定患者组织中miR-126的表达水平，并分析其表达与患者病理特征及患者预后的关系。将miR-126 inhibitor及其阴性对照（NC inhibitor）和miR-126 mimics及其阴性对照（NC mimics）分别转染至DLBCL细胞系SU-DHL-4中， RT-qPCR 检测细胞中miR-126的表达水平；MTT法检测细胞增殖活性；单克隆形成实验检测细胞集落形成能力；Annexin V/PI双染法检测细胞凋亡水平；Transwell实验检测细胞迁移及侵袭能力；Western blot法检测细胞凋亡相关蛋白cleaved-Caspase-3、Bcl-2和Bax的表达水平.
结果: miR-126在DLBCL患者淋巴瘤组织中高表达，其表达水平与DLBCL患者Hans分型、IPI评分和Ann-Arbor分期存在显著相关性（P<0.05）。Kaplan-Meier生存分析结果显示，miR-126高表达的DLBCL患者的生存率显著低于低表达患者（P<0.05）。与NC mimics组比较，miR-126 mimics组细胞中miR-126表达水平、细胞增殖率、集落数量、迁移和侵袭数量以及Bcl-2蛋白表达水平均明显增加（P<0.05），而细胞凋亡率、cleaved-Caspase-3和Bax蛋白表达量均明显降低（P<0.05）；与NC inhibitor组比较，miR-126 inhibitor组细胞中miR-126表达水平、细胞增殖率、集落数量、迁移和侵袭数量以及Bcl-2蛋白表达水平均明显降低（P<0.05），而细胞凋亡率、cleaved-Caspase-3和Bax蛋白表达水平均明显增加（P<0.05）.
结论: miR-126在DLBCL患者淋巴瘤组织中高表达，其表达水平与患者不良预后相关。miR-126可促进DLBCL细胞增殖、侵袭与迁移，并抑制细胞凋亡.
Keywords: apoptosis; diffuse large B-cell lymphoma; miR-126; prognosis; proliferation.