Jonas S, Windeatt S, O-Boateng A, Fordy C, Allen-Mersh T G
Department of Surgery, Charing Cross and Westminster Medical School, Chelsea and Westminster Hospital, London.
Gut. 1996 Nov;39(5):717-21. doi: 10.1136/gut.39.5.717.
Application of the reverse transcriptase polymerase chain reaction (RT-PCR) to identification of circulating tumour cells in colorectal cancer.
To assess whether circulating malignant cells in patients with colorectal liver metastasis could be identified by RT-PCR recognition of mRNA coding for the tumour marker carcinoembryonic antigen (CEA).
A total of 31 with colorectal liver metastases and 22 no-cancer controls.
Specific cDNA primers for CEA transcripts were used to apply RT-PCR to tissue biopsy specimens, colon carcinoma cell lines, and peripheral blood samples from patients with colorectal liver metastases. A strongly CEA-expressive HT115 colorectal carcinoma cell line was used to spike blood samples from no-cancer control subjects.
The limit for detection of CEA cDNA by Southern blotting using HT115 cells was 50 cells per 14 ml of spiked blood. There was a significant difference (p = 0.007) in RT-PCR positive expression between patients with liver metastasis (26/31) compared with controls (5/22). There was no significant relation between the prevalence of CEA cDNA amplification and serum CEA level or metastasis volume in patients with liver metastasis.
This is the first study to suggest that identification of circulating colorectal cancer cells using RT-PCR for detection of CEA cDNA is feasible.
逆转录聚合酶链反应（RT-PCR）在结直肠癌循环肿瘤细胞鉴定中的应用。
评估通过RT-PCR识别编码肿瘤标志物癌胚抗原（CEA）的mRNA，能否鉴定结直肠癌肝转移患者的循环恶性细胞。
31例结直肠癌肝转移患者和22例非癌症对照者。
使用针对CEA转录本的特异性cDNA引物，对结直肠癌肝转移患者的组织活检标本、结肠癌细胞系和外周血样本进行RT-PCR检测。使用高表达CEA的HT115结肠癌细胞系对非癌症对照者的血样进行加样。
使用HT115细胞通过Southern印迹法检测CEA cDNA的下限为每14 ml加样血中50个细胞。肝转移患者（26/31）与对照者（5/22）相比，RT-PCR阳性表达存在显著差异（p = 0.007）。肝转移患者中CEA cDNA扩增发生率与血清CEA水平或转移灶大小之间无显著相关性。
这是第一项表明使用RT-PCR检测CEA cDNA鉴定循环结直肠癌细胞是可行的研究。
