应用套式RT-PCR方法检测大肠癌患者外周血中的肿瘤细胞

Detection of tumor cells in circulating peripheral blood of colorec tal carcinoma patients by nested RT-PCR

目的 探讨应用套式RTPCR检测大肠癌患者外周血中的肿瘤细胞的可行性及其临床意义。方法 以CK20mRNA为靶基因，应用套式RTPCR方法检测10名健康人，17名非恶性消化系疾病患者，32名大肠癌患者手术前、后3天外周血中的肿瘤细胞。结果 CK20 mRNA在空白对照组中均呈阴性表达。大肠癌患者CK20表达情况为：2名Duke'sA期患者外周血中未检出CK20阳性细胞，15名Duke'sB期患者外周血中CK20mRNA阳性表达4例，10名Duke'sC 患者外周血CK20mRNA阳性表达6例，5名Duke'sD期患者外周血中CK20mRNA阳性表达4例。32名大肠癌患者中，术前CK20mRNA阳性总检出率为43.8%;A、B 期阳性率为23.5%；C、D期阳性率为66.7%。结论 以CK20为靶基因，应用套式RTPCR检测大肠癌患者外周血中的肿瘤细胞是一非损伤性且特异性、敏感性较高的检测方法。它将有助于肿瘤经血液微转移的早期诊断，并将有助于指导辅助化疗和监测其疗效。

：Objective Nested RT PCR was used to detect t umor cells cirulating peripheral blood of colorectal cancer patients and the cli nical significance was studied.Methods Total cellular RNA was e xtracted from whole blood of different groups：control group (10 healthy adults, 17 benign gastrointestinal disease patients), and test group (32 colorectal can cer patients). Reverse transtrcription and amplification were performed. Results CK 20 mRNA could not be detected in the whole blood of benign gastrointestinal disease patients and healthy adults.And CK 20 mRNA can also not be detected in the blood of 2 stage patients. But CK 20 mRNA can be detected in the blood of 4 out of 15 stage B patients, 6 out of 10 stage C patients, 4 out of 5 stage D patients.The total positive percentage of CK 20 mRNA was 43.8% in 32 colorectal cancer patients; the positive percentage of CK 20 mRNA in Stage A,B patients was 23.5% and in Stage C,D patients was 66.7%.Conclusion\ This test is a non invasive, sensitive and specific ass ay for detecting circulating epithelial disseminating disease.