凝血酶敏感蛋白-1在肺癌中的表达及临床意义

Expression and clinical significance of thrombin-sensitive protein-1 in lung cancer

目的探讨凝血酶敏感蛋白-1(TSP-1)在肺癌中的表达及临床意义。方法选取2016年9月至2019年12月间上海市第六人民医院收治的72例肺癌患者血清,根据TSP-1表达水平分组,患者分为TSP-1低表达组(41例)和正常表达组(31例)。采用qRT-PCR和Western blot检测细胞株中TSP-1 mRNA和蛋白表达,甲基化特异性PCR(MSP)检测TSP-1启动子区甲基化状态,ELISA检测肺癌患者血液中TSP-1蛋白表达。探讨TSP-1与临床病理因素及预后的相关性。结果qRTPCR结果显示,TSP-1 mRNA在肺细胞株A549和HCC827中的表达均低于肺正常细胞株(MRC-5),差异有统计学意义(P<0.05)。Western blot结果显示,TSP-1蛋白肺细胞株A549和HCC827表达低于肺癌细胞株肺正常细胞株(MRC-5),差异有统计学意义(P<0.05)。MSP检测发现,肺癌细胞株A549和HCC827中TSP-1基因启动子区发生甲基化,肺癌细胞株H1650未发现TSP-1基因甲基化,正常细胞株(MRC-5)中未发现TSP-1基因甲基化。TSP-1表达与临床病理因素分析,TSP-1表达与T分期和N分期均密切相关,差异有统计学意义(P<0.05)。预后单因素分析显示,肿瘤直径、T分期、N分期和TSP-1表达与患者预后相关,差异有统计学意义(P<0.05)。Cox预后多因素分析显示,N分期和TSP-1表达为患者预后的独立因素,差异有统计学意义(P<0.05)。进一步分析发现,TSP-1低表达患者预后较TSP-1正常表达患者预后差,差异有统计学意义(P<0.05)。结论TSP-1在肺癌中低表达,与启动子区甲基化密切相关,可作为肺癌预后预测因子。

Objective This study was to investigate the expression and clinical significance of thrombospondin-1(TSP-1)in lung cancer.Methods Blood serum was collected from 72 patients with lung cancer who received treatment at Shanghai Sixth People’s Hospital from September 2016 and December2019.They were categorized into a decreased expression of TSP-1 group(41 patients)and a normal expression of TSP-1 group(31 patients)based on the expression of TSP-1.The association between TSP-1 and pathological factors and prognosis was discussed.Results Real-time quantitative reverse transcription-polymerase chain reaction(qRT-PCR)test showed that the expression of TSP-1 mRNA was lower in lung cell line A549 than in normal lung cell line MRC-5(P<0.05).Western blot test indicated that the expression of TSP-1 protein was lower in lung cell line A549 than in normal lung cell line MRC-5(P<0.05).Methylation-specific PCR(MSP)detection suggested that the promoter region of TSP-1 gene was methylated in lung cancer cell line A549 and hcc827,while no TSP-1 gene promoter methylation was found in H1650 and MRC-5.An analysis of association between TSP-1 expression and pathological factors showed that the expression of TSP-1 was strongly associated with T stage and N stage(all P<0.05).Univariate analysis showed that tumor diameter,T stage and N stage and the expression of TSP-1 were closely associated with prognosis of lung cancer(all P<0.05).The multivariate Cox analysis of prognosis showed that N stage and the expression of TSP-1 were independent factors influencing the prognosis of lung cancer(all P<0.05).Moreover,the prognosis was poorer in patients with decreased expression of TSP-1 than in patients with increased expression of TSP-1.Conclusion The expression of TSP-1 was low in lung cancer which was mainly associated with methylation in the promoter region.TSP-1 could be a prognostic factor for lung cancer.