T7 RNA聚合酶催化荧光扩增技术检测结直肠癌患者外周血肿瘤细胞的临床意义

Detection of Circulating Tumor Cells in the Peripheral Blood of Patients with Colorectal Cancer by Fluorescent Amplification Catalyzed by T7 Polymerase Technique and Its Clinical Implications

目的:探讨T7RNA聚合酶催化荧光扩增技术检测结直肠癌患者术前外周血中循环肿瘤细胞的临床意义。方法:以CEA作为循环肿瘤细胞标志物,采用T7RNA聚合酶催化的荧光扩增技术,术前检测68例结直肠癌(结直肠癌组)及30例健康人(对照组)的外周血循环肿瘤细胞,并分析结直肠癌患者术前外周血中循环肿瘤细胞的检出率与临床病理特征的关系。结果:结直肠癌组总的循环肿瘤细胞检出率75.00%(51/68),57例无远处转移和11例有远处转移者检出率分别为70.20%(40/57)和100.00%(11/11)。循环肿瘤细胞在结直肠癌组DukesA、B、C、D期检出率分别为36.36%(4/11)、68.42%(13/19)、85.19%(23/27)和100.00%(11/11),DukesC+D期检出率明显高于DukesA+B期(P<0.01)。淋巴结转移者检出率为86.84%,而无转移者为60.00%,两者比较差异具有显著性统计学意义(P<0.05)。对照组出现1例假阳性,此方法的特异性为97%。结论:采用T7RNA聚合酶催化荧光扩增技术检测结直肠癌患者外周血循环肿瘤细胞检出率较高,其术前检出率与肿瘤分期和淋巴结转移密切相关,对结直肠癌早期诊断及判断预后具有重要意义。

Objective：To explore the detection of circulating tumor cells （CTCs）in the peripheral blood of patients with colorectal cancer ceils prior to surgery by a new method referred to as fluorescent amplification catalyzed by T7 polymerase technique and its clinical implications. Methods：Fluorescent amplification catalyzed by T7 polymerase technique was used to detect the CTCs prior to surgery by using CEA as tumor cell marker. 68 CRC patients and 11 cases with distant metastasis as well as 30 healthy subjects were examined; in addition, analysis was carried out for the positive rate of CTCs with patients＇ clinicopathologic features. Results： The overall positive incidence of CTCs was 75.00% （51/68）. Positive rate in patients with and without distant metastasis were 70.20% （40/57） and 100.00% （ 11/11 ） respectively. 36.36% （4/11） ,68.42% （13/19） ,85.19% （23/27） and 100. 00% （ 11/11 ） of CRC patients were positive for Dukes A, B, C, D respectively. The positive rate of CTCs in Dukes C and D were higher than that in Dukes A and B （ P 〈 0. 01 ）. CTCs detection in patients of lymph node metastasis positive group（86.84% ）was significantly higher than the negative group（60. 00% ） （P 〈 0.05 ）. One fake positive case was found in the control group and its specificity was 97%. Conclusion：Fluorescent amplification catalyzed by T7 polymerase technique can he used to detect CTCs in the peripheral blood of CRC patents. The positive rate of CTCs was significantly related to Dukes stage and lymph node metastasis status. Detection of CTCs is beneficial to early diagnosis of CRC and prediction of prognosis.