Fig 1.
Fluorescence microscopy images of cells isolated from peripheral blood of HCC patients by positive immunomagnetic separation.
Images of cells stained with DAPI (blue), anti-cytokeratin antibodies (green), and CD45 antibodies (red) and the corresponding merged image are shown. Cytokeratin-positive cells are CTCs, whereas CD45-positive cells are lymphocytes.
Fig 2.
The first step represents the forward-scatter (FSC) versus side-scatter (SSC) plot (left). In this step, the unassigned events and artifacts (including cell debris, mechanical noise, and cell clumps) are gated off. In the second step, the dead cells and CD235a-positive cells are gated off on the CD235a versus 7-AAD plot (middle). In the remaining CD45-negative cells, GPC3-positive cells are selected with the CD45 versus GPC3 plot (right).
Fig 3.
Linear correlation between the numbers of spiked and detected HepG2 cells.
HepG2 cells (10, 100, 250, and 500 cells) were spiked into 8.0 mL of blood from a healthy volunteer. The number of cells spiked (x-axis) is plotted versus the number of cells detected by flow cytometry (y-axis), with the correlation coefficient shown at the top of the graph.
Fig 4.
Dot and box-plots of GPC3-positive CTC count.
Distribution of GPC3-positive CTCs in HCC patients (n = 85), healthy volunteers (n = 12), cirrhotic patients (n = 4), patients with benign liver tumors (n = 4), patients with malignant liver tumors other than HCC (n = 5), and patients with colorectal cancer (n = 2).
Fig 5.
(A) ROC curve of CTC count for discrimination between HCC patients and non-HCC patients. (B) ROC curve of CTC count for prediction of microscopic portal vein invasion in HCC patients.
Table 1.
Clinicopathologic features.
Table 2.
Clinicopathologic features between CTC ≥5 and <5 group.
Table 3.
Preoperative factors associated with microscopic portal vein invasion.
Fig 6.
(A) Comparison of disease-free survival rates after hepatectomy of the CTC ≥5 group to that of the CTC <5 group. The disease-free survival rates were significantly lower in the CTC ≥5 group than the CTC <5 group (log-rank test; p = 0.015). (B) Comparison of overall survival rates after hepatectomy of the CTC ≥5 group to that of the CTC <5 group. The overall survival rates were significantly lower in the CTC ≥5 group than the CTC <5 group (log-rank test; p = 0.047).