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利用逆转录和聚合酶链反应检测乳腺癌患者术中肿瘤细胞播散

Detection of intraoperative tumor cell dissemination in patients with breast cancer by use of reverse transcription and polymerase chain reaction.

作者信息

Brown D C, Purushotham A D, Birnie G D, George W D

机构信息

University Department of Surgery, Western Infirmary, Glasgow, Scotland.

出版信息

Surgery. 1995 Jan;117(1):95-101. doi: 10.1016/s0039-6060(05)80235-1.

Abstract

BACKGROUND

Animal studies have shown that malignant cells are shed into the blood stream during surgical resection of a primary tumor and that this may enhance the development of metastases. The evidence for tumor cell dissemination during surgical manipulation of human cancer is unclear. We have applied the technique of reverse transcription and polymerase chain reaction to detect circulating tumor cells in peripheral venous blood of patients with breast cancer perioperatively.

METHODS

To target breast-specific gene transcription complementary DNA was prepared by reverse transcription of blood messenger RNA with oligonucleotide primers unique to CK18 and DF3 antigens. Preliminary assessment of specificity showed that the DF3 antigen was more suitable than CK18 for the purpose of this study. Assessment of sensitivity showed that as few as 10 tumor cells per 5 ml blood could be identified by this method. Peripheral blood samples were obtained by venepuncture from patients before, during, and 24 hours after breast surgery (nine malignant and three benign).

RESULTS

In the group of patients with malignant disease, tumor cells were detected in one patient before operation and four patients during operation. No tumor cells were detected in the postoperative samples nor in any of the samples of patients with benign disease.

CONCLUSIONS

These findings suggest that tumor manipulation during operation encourages tumor cell dissemination.

摘要

背景

动物研究表明,在原发性肿瘤手术切除过程中恶性细胞会进入血流,这可能会促进转移灶的形成。关于人类癌症手术操作过程中肿瘤细胞播散的证据尚不清楚。我们应用逆转录和聚合酶链反应技术对乳腺癌患者围手术期外周静脉血中的循环肿瘤细胞进行检测。

方法

为靶向乳腺特异性基因转录,用针对细胞角蛋白18(CK18)和DF3抗原的独特寡核苷酸引物对血液信使核糖核酸进行逆转录制备互补脱氧核糖核酸。特异性的初步评估显示,DF3抗原比CK18更适合本研究目的。敏感性评估显示,该方法可识别出每5毫升血液中低至10个肿瘤细胞。通过静脉穿刺从乳腺癌手术前、手术中和术后24小时的患者(9例恶性肿瘤患者和3例良性疾病患者)获取外周血样本。

结果

在恶性疾病患者组中,术前1例患者和术中4例患者检测到肿瘤细胞。术后样本以及良性疾病患者的任何样本中均未检测到肿瘤细胞。

结论

这些发现提示手术过程中的肿瘤操作会促使肿瘤细胞播散。

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