食管癌围手术期外周血微转移的定量检测及其临床意义

Detection of micrometastasis in perioperative primary esophageal cancer patients using real-time quantitative reverse transcrtption-PCR

目的探讨食管癌围手术期外周血微转移定量检测的理想途径,判断手术对微转移的影响程度。方法对40例可手术的食管癌分别于手术前1天(记为B-1)、手术刚结束时(记为B0)和手术后第3天(记为B+3)取外周中心静脉血,分离单核细胞并提取总RNA,用实时荧光PCR技术定量检测癌胚抗原基因(CEAmRNA)表达水平,以胸部良性肿瘤患者和正常人为对照。结果CEA mRNA拷贝值在B-1、B0和B+3取样时间点的中位值分别为1592、13 314、6221拷贝/ml。且在B0及B+3处的CEA mRNA表达水平都明显高于在B-1处的表达水平(P值分别为0.0001和0.0209)。CEA mRNA表达水平在B0处高于B+3处,但差异无统计学意义(P=0.4396)。结论手术操作使得患者外周血的CEA mRNA表达水平明显增加,术后第3天有部分下降,但仍有部分患者存在高水平的CEA mRNA,表明手术促进微转移的发生,有必要行围手术期综合治疗。

Objective To establish a desirable quantitative assay system to evaluate the correlation between surgical manipulation and micrometastasis in primary esophageal cancer patients.Methods 118 pe- ripheral blood samples from 40 esophageal cancer patients undergoing radical resection were collected before surgery(B-1),immediately after surgery(B0)and at the third day postoperatively(B+3).Based on real-time quantitative reverse transcription-PCR,carcinoembryonic antigen(CEA)mRNA gene was used in the detec- tion.12 patients with benign tumor and 13 healthy volunteers were collected as negative control groups.Re- sults The median of CEA mRNA copies at B-1,B0 and B+3 were 1592,13 314 and 6221 copies/ml blood, respectively.CEA mRNA expression levels were found to be significantly higher at both B0 and B+3 than that of B-1(P=0.0001 and 0.0209,respectively).No significant difference was found between B0 and B+3(P= 0.4396).Conclusion Surgical manipulation on esophageal cancer patients increases the probability of mi- crometastasis.Therefore,adjuvant therapy is needed during perioperative stage.