Detection of apoptotic circulating tumor cells in advanced pancreatic cancer following 5-fluorouracil chemotherapy
Detection of apoptotic circulating tumor cells in advanced pancreatic cancer following 5-fluorouracil chemotherapy
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5-氟尿嘧啶化疗后晚期胰腺癌循环肿瘤细胞凋亡的检测
DOI：
10.4161/cbt.12.8.15960
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发表时间：
2011-10-15
影响因子：
3.6
通讯作者：
Zhao, Xiaohang
中科院分区：
文献类型：
--
作者：
Ren, Chuanli;Han, Chongxu;Zhao, Xiaohang
本研究旨在探讨CA19-9和CK8/18在5-氟尿嘧啶(5-FU)诱导的胰腺癌细胞株和未经治疗的晚期胰腺癌患者外周血中肿瘤细胞(CTCs)中的表达模式。此外，我们的目标是测试CTCs的动态变化与第一周期化疗效果的关系。为了完成这项研究，CD45抗体包被的珠子被用来丢弃外周血中的白细胞。结合CA19-9-Alexa488和CK8/18-Alexa594免疫荧光染色对41例晚期胰腺癌患者化疗前后循环中的CTCs进行鉴定。将PL45胰腺癌细胞与20µm ol/L的5-FU孵育12 h和24 h，检测CA19-9和CK8/18的表达模式，评价细胞凋亡的程度。随后，检测细胞凋亡率和CTCs。在41例Ⅲ期和Ⅳ期胰腺癌患者中，80.5%的患者治疗前外周血中存在两个以上的CTC，其中位数为16.8±16.0(0~59)。第1周期5-FU化疗7天后，仅29.3%(12/41)患者外周血中CTCs>2个，中位数为3.8±7.8(0~40个)(P=0.000)。20例健康献血员外周血7.5ml中未检出CTCs(P=0.000)。在晚期胰腺患者接受第一周期化疗后，检测到凋亡的CTCs。CA19-9和CK8/18染色可见点状、颗粒状和泡状形态，可能反映胰腺癌细胞和CTCs的凋亡。凋亡的CTCs可能预示着胰腺癌患者化疗的疗效。然而，还需要进一步的研究来跟进这些发现。
The objective of the present study was to evaluate CA19-9 and CK8/18 expression patterns in pancreatic cancer cell lines induced by 5-fluorouracil (5-Fu), and in circulating tumor cells (CTCs) in peripheral blood of patients previously untreated with advanced pancreatic cancer. Furthermore, the goal was to test the relationship of dynamic of CTCs with the effects of the first cycle of chemotherapy. To accomplish this study, CD45 antibody coated beads were used to discard white blood cells in peripheral blood. This was done in combination with CA19-9-Alexa488 and CK8/18-Alexa594 immunofluorescence staining to identify CTCs in circulation from 41 advanced pancreatic cancer patients, before and after chemotherapy. The PL45 pancreatic cancer cell line was incubated with 20 µmmol/L 5-Fu for 12 and 24 hours to induce apoptosis, and the expression patterns of CA19-9 and CK8/18 were measured, and the extent of apoptosis was evaluated. Subsequently, apoptotic cells and CTCs were measured. Of 41 patients with stage III and Ⅳ pancreatic cancer, 80.5% were detected with more than two CTCs in 7.5mL peripheral blood before any therapy and the median number of CTCs was 16.8±16.0 (0-59). After 7 days by the first cycle of 5-Fu chemotherapy, only 29.3% (12/41) of these patients were detected more than two CTCs in 7.5mL peripheral blood and the median number of CTCs was 3.8±7.8 (0-40) (P=0.000). And no CTCs were detected in 20 healthy donors from 7.5mL peripheral blood (P=0.000). Apoptotic CTCs were detected after advanced pancreatic patients were administered the first cycle of chemotherapy. Punctate, granular, and bubble-like morphologies with CA19-9 and CK8/18 staining were found and may reflect apoptosis in pancreatic cancer cells and CTCs. Apoptotic CTCs may indicate the efficacy of chemotherapy in pancreatic cancer patients. However, further studies are required to follow up these findings.