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目的探讨采用巢式逆转录聚合酶链反应(RT-PCR)检测胃癌患者腹腔冲洗液中癌胚抗原(CEA)mRNA和细胞角蛋白20(CK-20)mRNA的临床价值。方法收集80例胃癌患者和10例胃良性病变患者的腹腔冲洗液或腹水,常规行HE染色细胞学检查,采用巢式RT-PCR方法检测腹腔冲洗液中CEA mRNA和CK-20 mRNA的表达,并以人胃癌细胞株SGC-7901作为阳性对照。结果80例胃癌患者腹腔冲洗液中CEA mRNA的阳性表达率为52.5%(42/80),CK-20 mRNA的阳性表达率为57.5%(46/80),CEA mRNA和CK-20 mRNA的联合检测阳性率为70.0%(56/80),皆高于腹腔冲洗液细胞学(PLC)检查的27.5%(22/80,P<0.05),且22例PLC检查阳性患者的CEA mRNA和CK-20 mRNA检测结果均为阳性。CEA mRNA和CK-20 mRNA的阳性率与淋巴结转移和TNM分期明显相关(P<0.05)。结论巢式RT-PCR检测胃癌患者腹腔冲洗液中CEA和CK-20基因,可以提高腹腔内游离癌细胞(IFCCs)的检测灵敏度和特异度,对于早期诊断胃癌腹膜微转移有一定的临床价值,可为手术方式选择及术中、术后化疗提供依据。
Objective To investigate the clinical value of nested reverse transcriptase polymerase chain reaction (RT-PCR) in the detection of carcinoembryonic antigen (CEA) mRNA and cytokeratin 20 (CK-20) mRNA in peritoneal washings of gastric cancer patients. Methods Peritoneal washings and ascites were collected from 80 patients with gastric cancer and 10 patients with benign gastric lesions. HE staining and cytology were performed routinely. The expressions of CEA mRNA and CK-20 mRNA in peritoneal washings were detected by nested RT-PCR. Human gastric cancer cell line SGC-7901 was used as a positive control. Results The positive expression rate of CEA mRNA in peritoneal washings was 52.5% (42/80) in 80 gastric cancer patients and 57.5% (46/80) in CK-20 mRNA. The combination of CEA mRNA and CK-20 mRNA The detection positive rate was 70.0% (56/80), which was higher than that of PLC (27.5%, 22/80, P <0.05), and the positive rate of CEA mRNA and CK- 20 mRNA test results were positive. The positive rates of CEA mRNA and CK-20 mRNA were significantly correlated with lymph node metastasis and TNM staging (P <0.05). Conclusion The detection of CEA and CK-20 gene in peritoneal washings by nested reverse transcription-polymerase chain reaction (RT-PCR) can improve the detection sensitivity and specificity of intraperitoneal free carcinoma cells (IFCCs), which has some clinical value for the early diagnosis of peritoneal micrometastasis in gastric cancer. For surgical options and intraoperative and postoperative chemotherapy to provide the basis.