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目的建立高效、快速的外周血Cytokeratin 20(CK20)表达的检测方法,探讨大肠癌患者外周血中CK20的表达与微转移的关系,研究大肠癌与p53基因突变的关系。方法运用荧光监测技术结合RT-PCR,检测36例大肠癌患者术前和术后不同时间外周血中的CK20的表达。用SSCP技术对大肠癌患者进行p53基因的突变检测。结果36例患者术前的外周血中检出CK20表达的有27例,阳性率为75%,手术后24 h患者外周血中检出CK20表达的有30例,阳性率为83.3%,术后早期化疗结束后仅13例为阳性,阳性率为36%。36例患者的p53基因的SSCP突变检测中未检测到突变。结论运用荧光RT-PCR技术检测外周血中CK20的表达可以监测癌症微转移的发生,简便快速,检出率高。
Objective To establish an efficient and rapid method for the detection of Cytokeratin 20 (CK20) expression in peripheral blood, to investigate the relationship between the expression of CK20 and the micrometastasis in patients with colorectal cancer, and to investigate the relationship between colorectal cancer and p53 gene mutation. Methods The expression of CK20 in peripheral blood of 36 patients with colorectal cancer before and after operation was detected by fluorescence monitoring and RT-PCR. SSCP was used to detect the mutation of p53 gene in patients with colorectal cancer. Results Twenty-seven cases of CK20 were found in the peripheral blood of the preoperative patients. The positive rate was 75%. Twenty patients with CK20 were detected in peripheral blood at 24 hours after surgery. The positive rate was 83.3%. Only 13 cases were positive after the end of early chemotherapy, and the positive rate was 36%. No mutations were detected in the SSCP mutations of the p53 gene in 36 patients. Conclusion The use of fluorescent RT-PCR to detect the expression of CK20 in peripheral blood can monitor the occurrence of cancer micrometastasis, which is simple and rapid, and the detection rate is high.