Objective To investigate the expression of sperm-associated antigen 6 (Spag6) in liver cancer tissue and its association with the clinicopathological features and prognosis of liver cancer patients,as well as the effect of Spag6 on the proliferation and migration of HCCLM3 hepatoma cells.Methods Clinical samples were collected from 102 liver cancer patients who were treated in Xiangya Hospital of Central South University from August 2006 to November 2009,and Western blot was used to measure the expression of Spag6 in hepatoma cells,normal liver tissue,tumor tissue,and corresponding adjacent tissue.Immunohistochemistry was used to measure the expression of Spag6 in 102 liver cancer tissue samples,and according to the immunohistochemical scoring criteria,the patients were divided into high Spag6 expression group and low Spag6 expression group.Lentivirus-mediated RNA interference technique was used to silence Spag6 expression in HCCLM3 cells;Western blot was used to analyze silencing effect,wound-healing assay was used to investigate the effect of Spag6 gene silencing on the migration of HCCLM3 cells,and colony formation assay was performed to observe the effect of Spag6 gene silencing on the proliferation of HCCLM3 cells.The chi-square test was used to investigate the association between Spag6 expression and clinicopathological features of liver cancer patients,and the Kaplan-Meier survival analysis and log-rank test were used to analyze the association between Spag6 expression and the prognosis of liver cancer patients.Results Hepatoma cells and liver cancer tissue had significantly higher expression of Spag6 than the normal L02 cells and normal liver tissue.Immunohistochemistry showed that the expression rate of Spag6 was 58.8％ (60/102) in liver cancer tissue samples and 12.7％ (13/102) in adjacent tissue samples (x2 =47.123,P ＜0.001).According to the results of the chi-square test,Spag6 expression was associated with the number of tumor nodules,presence or absence of capsule,vascular invasion,and Edmondson-Steiner classification (x2 =8.360,6.761,4.344,and 7.172,P =0.004,0.009,0.037,and 0.007).Further analysis showed that the high Spag6 expression group had significantly lower 1-,3-,and 5-year survival rates than the low Spag6 expression group (71.5％ vs 90.5％,43.7％ vs 68.8％,19.7％ vs 48.7％,x2 =11.228,P =0.001).Cell assays showed significant reductions in the proliferation and migration of HCCLM3 cells after Spag6 gene silencing (both P ＜ 0.01).Conclusion Spag6 is highly expressed in hepatoma cells and liver cancer tissue,and its high expression is associated with poor clinicopathological features and postoperative survival of liver cancer.Spag6 can promote the proliferation and migration of hepatoma cells,suggesting that Spag6 may be involved in the development and progression of liver cancer.Therefore,it can be used as a reference index for predicting the prognosis of liver cancer patients and a potential target for liver cancer treatment.%目的 检测精子相关抗原6(Spag6)在肝癌组织中的表达情况及其与肝癌患者临床病理特征和预后的关系,探讨Spag6对肝癌细胞HCCLM3增殖和迁移的影响.方法 选取2006年8月-2009年11月来自中南大学湘雅医院的102例肝癌患者组织样本进行研究,应用Western Blot检测肝癌细胞系、正常肝组织、肿瘤组织及对应的癌旁组织中Spag6表达水平.利用免疫组化法检测102例肝癌组织中Spag6的表达,并根据肿瘤组织免疫组化评分标准分成Spag6高表达组和低表达组.用慢病毒介导的RNAi干扰技术沉默Spag6基因在HCCLM3细胞中的表达,Western Blot检测沉默效果;利用细胞划痕愈合实验检测沉默Spag6基因后对HCCLM3细胞迁移的影响,集落形成实验观察对细胞增殖的影响.采用x2检验分析Spag6表达水平与肝癌患者临床病理特征的关系;利用Kaplan-Meier生存分析、log-rank检验分析Spag6表达水平与肝癌患者预后的关系.结果 Spag6在肝癌细胞系和肝癌组织中表达高于正常LO2细胞以及正常肝组织.免疫组化结果显示102例标本中肝癌组织的Spag6的表达率为58.8％ (60/102),癌旁组织中Spa6的表达率为12.7％(13/102),差异有统计学意义(x2=47.123,P＜0.001).Spag6的表达水平与肿瘤结节数目、有无包膜、血管侵犯、Edmondson-Steiner分级有关(x2值分别为8.360、6.761、4.344、7.172,P值分别为0.004、0.009、0.037、0.007).进一步研究显示Spag6高表达组的1、3、5年生存率明显低于低表达组(71.5％ vs 90.5％,43.7％ vs 68.8％,19.79％ vs48.7％∥=11.228,P=0.001).细胞实验证实沉默Spag6基因后HCCLM3细胞增殖和迁移能力明显减弱(P值均＜0.01).结论 Spag6在肝癌细胞及肝癌组织中均呈高表达,且其高表达与肝癌不良临床病理特征以及术后生存有关;Spag6能够促进肝癌细胞增殖和迁移运动,提示Spag6参与了肝癌的发生发展,可作为判断肝癌患者预后的参考指标以及治疗的可能靶点.
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