Clinical significance of pancreatic circulating tumor cells using combined negative enrichment and immunostaining-fluorescence in situ hybridization

Abstract

BackgroundCirculating tumor cells (CTCs) hold great potential in both clinical application and basic research for the managements of cancer. However, it remains to be an enormous challenge to obtain efficient detection of pancreatic CTCs. New detection platforms for the detection of pancreatic CTCs are urgently required.MethodsIn the present study, we applied a newly-developed platform integrated subtraction enrichment and immunostaining-fluorescence in situ hybridization (SE-iFISH) to analyze clinical significance of pancreatic CTCs. Immunostaining of CK, CD45, DAPI and FISH with the centromere of chromosome 8 (CEP8) were utilized to identify CTCs. Cells with features of CK+/CD45-/DAPI+/CEP8 = 2, CK+/CD45-/DAPI+/CEP8 > 2, CK-/CD45-/DAPI+/CEP8 > 2 were defined as pancreatic CTCs. The Kaplan-Meier method and Cox proportional hazards model were used to analyze the relationship of CTC level and other clinicopathological factors with pancreatic cancer clinical outcomes.ResultsCTC count in pancreatic cancer was higher than healthy individuals (median, 3 vs 0 per 7.5 ml; P < 0.001). SE-iFISH platform yielded a sensitivity of 88 % and specificity of 90 % in pancreatic cancer at the cutoff value of 2 cells/7.5 ml. Pancreatic cancer patients with lower CTC count (<3/7.5 ml) had substantially better overall survival (OS) compared with these with higher CTC count (≥3/7.5 ml) (15.2 vs 10.2 months, P = 0.023). Multivariate analysis indicated that higher CTC count was a strong indicator for worse OS (HR = 4.547, P = 0.016).ConclusionOur current data showed that CTCs could be detected in pancreatic cancer patients in various stages, whether localized, locally advanced and metastatic. Besides, CTCs have shown the potential implication in predicting prognosis of pancreatic cancer.

Circulating Tumor Cells and Circulating Tumor DNA Provide New Insights into Pancreatic Cancer

“…Several modified device have been developed for better detection of pancreatic CTCs. Immuno-FISH platform is a negative-enrichment method for CTC detection and our preliminary results showed that the sensitivity could reach 100% by combining CTC and CA19-9 36. The PCR-based strategy have also been reported to detect pancreatic CTCs, but the platform may produce false-positive results 37, 38.…”

Clinical significance of circulating tumor cells via combined whole exome sequencing in early stage cancer screening: A case report

“…These studies demonstrated SE-iFISH CTC detection 91% positive rate for breast cancer patients (7), a sensitivity of 88% and specificity of 90% in pancreatic cancer and healthy individuals at the cutoff value of 2 cells/7.5 ml (8), and a sensitivity of 90.9% and specificity of 82.4% in colorectal cancer and healthy individuals at the cutoff value of one cell/7.5 ml (9). From these studies, CTC detection by SE-iFISH showed high sensitivity and specificity for distinguishing breast cancer, pancreatic cancer and colorectal cancer from healthy people.…”

Circulating tumor cells and miRNAs as prognostic markers in neuroendocrine neoplasms

“…At the same time, Cell Search system has an inherent limitation that hampers the detection of EpCAM-negative CTCs. This drawback is overcome by a negative enrichment method (CD45−) combined with immunostaining-fluorescence ISH, which is able to yield CTCs independently of EpCAM expression and cell size (Gao et al 2016). A possible limitation, in case of earlystage cancer, is the difficulty to count CTCs as they are expected to be few, corresponding to a low sensitivity of the method.…”