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目的探讨轴突导向因子1(Netrin-1)在多发性骨髓瘤(MM)中的表达及其与患者临床病理特征的相关性。方法回顾性研究选取MM患者的骨髓标本40例作为研究组(MM组),另选同期骨髓象正常的门诊患者10例作为对照组。取患者治疗前、治疗后以及健康对照组的骨髓液,使用Ficoll密度梯度离心法从骨髓抽吸物中分离出单核细胞(MC),培养48 h后,应用免疫细胞化学检测细胞中Netrin-1的表达情况,应用蛋白免疫印迹检测细胞中Netrin-1、DCC、UNC5H蛋白表达情况。比较治疗无效与治疗有效组Netrin-1表达水平,分析Netrin-1蛋白表达水平临床特征的相关性,以及Netrin-1表达水平与结肠癌缺失蛋白(DCC)、UNC5同源蛋白(UNC5H)蛋白的相关性分析。结果与对照组患者MC相比,MM组患者MC免疫细胞染色显示Netrin-1表达增加。蛋白免疫印迹结果显示,与对照组相比,MM组患者MC中Netrin-1蛋白表达水平显著增加,差异有统计学意义(P<0.05)。与治疗前MM患者相比,治疗有效组MM患者Netrin-1蛋白表达水平显著降低,差异有统计学意义(P<0.05)。MM患者中Netrin-1蛋白表达水平与性别、年龄、免疫分型等临床特征无明显相关性(P>0.05),而与ISS分期、D-S分期等显著相关(P<0.05)。与对照组相比,MM组患者DCC、UNC5H蛋白表达水平均显著降低,差异均有统计学意义(P<0.05)。相关性分析显示,Netrin-1与DCC呈显著负相关(r=-0.4058,P=0.0094),而与UNC5H无显著相关性(r=0.0055,P=0.0893)。结论Netrin-1的表达与MM的发生发展密切相关,可能成为肿瘤生物治疗中潜在的治疗靶点。
Objective To investigate the expression of Netrin-1 in multiple myeloma (MM) and its relationship with clinicopathological features. Methods A retrospective study selected 40 patients with bone marrow samples of patients with MM as the study group (MM group), another 10 patients with normal bone marrow as the control group. Mononuclear cells (MCs) were isolated from bone marrow aspirate before and after treatment, and after treatment, and in healthy control group. After cultured for 48 hours, immunocytochemistry was used to detect the expression of Netrin- 1 expression, Western blotting was used to detect the expression of Netrin-1, DCC and UNC5H in the cells. The levels of Netrin-1 protein were compared between the treatment-ineffective and therapeutically-effective groups, and the correlation between Netrin-1 expression and colon cancer undecided protein (DCC), UNC5 homologous protein (UNC5H) Correlation analysis. Results Compared with control group MC, the staining of MC immunocytes in MM group showed that the expression of Netrin-1 was increased. Western blot results showed that compared with the control group, the expression of Netrin-1 protein in MC group was significantly increased (P <0.05). Compared with MM patients before treatment, the expression of Netrin-1 protein in MM group was significantly decreased (P <0.05). There was no significant correlation between the expression of Netrin-1 protein and clinical features such as gender, age and immunophenotype in MM patients (P> 0.05), but significantly correlated with ISS stage and D-S stage (P <0.05). Compared with the control group, the expression levels of DCC and UNC5H in MM group were significantly decreased (P <0.05). Correlation analysis showed that there was a significant negative correlation between Netrin-1 and DCC (r = -0.4058, P = 0.0094), but not with UNC5H (r = 0.0055, P = 0.0893). Conclusion The expression of Netrin-1 is closely related to the occurrence and development of MM, and may be a potential therapeutic target in tumor biological therapy.