术中实时RT-PCR检测胃癌患者腹腔冲洗液癌胚抗原mRNA的初步研究

Clinical significance of rapid quantitative detection of CEA mRNA levels in peritoneal washes from gastric cancer patients with real-time RT-PCR during operation

目的探讨术中实时逆转录聚合酶链反应(RT-PCR)定量检测胃癌腹腔冲洗液癌胚抗原信使核糖核酸(CEAmRNA)诊断腹腔游离癌细胞的可行性、可靠性及其临床意义。方法采用实时RT-PCR技术检测98例胃癌患者手术前后腹腔冲洗液CEAmRNA水平,并与常规RT-PCR、细胞学检查结果进行比较分析。结果全组腹腔冲洗液细胞学检查(PLC)阳性率为38.8%(38/98),常规RT-PCRCEAmRNA检测阳性率为52.0%(51/98),两者差异有显著性意义(P<0.01)。腹腔冲洗液CEAmRNA浓度术前为犤172.11±63.07(范围0～5.623×106)犦拷贝(copies)/ml,术后为犤606.32±76.21(范围2.564×102～4.677×107)犦copies/ml,术后显著高于术前(P<0.01)。肿瘤侵犯浆膜,病期Ⅲ、Ⅳ期,细胞学检查及RT-PCRCEAmRNA阳性者腹腔冲洗液CEAmRNA水平显著升高(均P<0.01),而淋巴结转移与否及肿瘤细胞分化程度不同组之间的腹腔冲洗液CEAmRNA水平比较,差异无显著性意义(均P>0.05)。PLC(-)PCR(-)组、PLC(-)PCR(+)组和PLC(+)PCR(+)组3组间腹腔冲洗液CEAmRNA浓度比较,差异有显著性意义(均P<0.05)。本研究将判断腹腔游离癌细胞的CEAmRNA界值初步定为21.73copies/ml。结论实时RT-PCR定量检测胃癌患者腹腔冲洗液CEAmRNA,是诊断腹腔游离癌细胞快速、敏感、可行的方法。

Objective To evaluate the feasibility and reliability of real time RT PCR measurement of carcinoembryonic antigen(CEA) mRNA levels in peritoneal washes of gastric cancers for detection of free tumor cells. Methods Peritoneal washes were obtained from 98 patients with gastric carcinoma before and after tumor resections. CEA mRNA levels were quantified using a real time PCR system with fluorescent hybridization probes. The results of real time PCR detection were compared with those of peritoneal lavage cytology(PLC)and routine RT PCR detection of CEA mRNA. Results The positive rate of PLC and routine RT PCR were 38.8%(38/98) and 52.0%(51/98)respectively,and the difference was significant (P< 0.01). The mean CEA mRNA level in peritoneal washes was [172.11±63.07(range 0 to 5.623×106)]copies/ml before resection, and [606.32±76.21(range 2.564×102 to 4.677×107)]copies/ml after resection(P< 0.01). The CEA mRNA levels in peritoneal washes of gastric cancer patients with tumor serosal invasion,Ⅲand Ⅳstages,positive PLC and RT PCR CEA mRNA detection were significantly higher(P< 0.01). The CEA mRNA levels were not related to lymph node metastasis and tumor cell differentiation (P >0.05),while/but were significantly different among the PLC(-)PCR(-)group,PLC(-)PCR(+)group,and PLC(+)PCR(+)group(all P< 0.05). The cut off value of CEA mRNA level for tumor free cells detection in peritoneal washes was 21.73 copies/ml in this current study. Conclusion Rapid quantitative detection of CEA mRNA with real time RT PCR is a rapid, sensitive and feasible method to examine free cancer cells in peritoneal washes of gastric cancer patients.