Department of Surgical Oncology, University of Tokyo, Tokyo, Japan.
Cytometry B Clin Cytom. 2014 Jan;86(1):56-62. doi: 10.1002/cyto.b.21126. Epub 2013 Sep 30.
Peritoneal metastasis (PM) is the most life-threatening type of metastasis in abdominal malignancy. To improve the diagnostic accuracy of cytologic detection (CY) of free tumor cells (FTC) in the peritoneal cavity, we tried to quantify the FTC to leukocyte ratio using flow cytometry in patients with peritoneal metastasis.
Cells were recovered from ascites or peritoneal lavages from 106 patients who underwent abdominal surgery and additional 89 samples which were obtained from peritoneal catheter or access port in patients with PM (+) gastric cancer. The cells were immunostained with monoclonal antibodies to CD45 and to CD326 (EpCAM). Using flow cytometry, CD326 (+) and CD45 (+) cells were classified as either tumor cells (T) or leukocytes (L) and the T/L ratio (TLR) was calculated.
In 106 samples obtained by laparotomy, Median (M) of the TLR of PM (+) patients was 1.39% (0-807.87%) which was significantly higher than PM (-) patients (M=0%, 0-2.14%, P < 0.001). In PM (+) patients, 86 CY (+) samples showed higher TLR than 61 CY (-) samples (M=2.81%, 0.02-1868.44% vs. M=0%, 0-3.45%, p<0.0001). In all of the 24 patients who were monitored for TLR before and after intraperitoneal (IP) chemotherapy, the TLR was reduced which was more dramatic than the results of the change in cytology.
TLR measured with FACS is an excellent reflection of the tumor spread in the peritoneal cavity and could be a reliable diagnostic biomarker to determine the severity of PM as well as effectiveness of IP chemotherapy.
腹膜转移(PM)是腹部恶性肿瘤中最具威胁生命的转移类型。为了提高腹腔游离肿瘤细胞(FTC)细胞学检测(CY)的诊断准确性,我们尝试使用流式细胞术定量分析腹膜转移患者腹腔内 FTC 与白细胞的比值。
从 106 例行腹部手术的患者的腹水或腹腔灌洗液中以及 89 例 PM(+)胃癌患者的腹膜导管或接入端口中获取细胞。用单克隆抗体 CD45 和 CD326(EpCAM)对细胞进行免疫染色。使用流式细胞术,将 CD326(+)和 CD45(+)细胞分为肿瘤细胞(T)或白细胞(L),并计算 T/L 比值(TLR)。
在剖腹术获得的 106 个样本中,PM(+)患者的 TLR 中位数(M)为 1.39%(0-807.87%),显著高于 PM(-)患者(M=0%,0-2.14%,P<0.001)。在 PM(+)患者中,86 例 CY(+)样本的 TLR 高于 61 例 CY(-)样本(M=2.81%,0.02-1868.44% vs. M=0%,0-3.45%,p<0.0001)。在所有 24 例接受腹腔内(IP)化疗前后 TLR 监测的患者中,TLR 均有所降低,且比细胞学变化更明显。
FACS 测量的 TLR 是对腹腔内肿瘤扩散的极好反映,可作为一种可靠的诊断生物标志物,用于确定 PM 的严重程度以及 IP 化疗的效果。