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Analysis of circulating tumor cells in colorectal cancer liver metastasis patients before and after cryosurgery

View Article: PubMed Central - PubMed

ABSTRACT

In this study, we determined the number of peripheral blood circulating tumor cells (CTCs) pre- and post-cryosurgery in patients with colorectal cancer liver metastasis as a reference for understanding the relevance of any changes to the efficacy of cryosurgery. CTC numbers and CTC-related gene expression were measured in the peripheral blood of 55 patients with colorectal liver metastasis at 1 day before and 7 and 30 d after cryoablation using magnetic activated cell sorting (MACS) and fluorescence activated cell sorting (FACS) combined with real-time quantitative PCR (RT-qPCR). The number of CTCs decreased significantly with postoperative time (P < 0.01). Delta cycle threshold values for the CTC-related genes CEA, Ep-CAM, CK18 and CK19 increased significantly after cryoablation. Furthermore, the expression of CEA, Ep-CAM, CK18 and CK19 decreased significantly with time after cryoablation (P < 0.01). RT-qPCR and FACS combined with MACS has significant diagnostic and prognostic value for evaluating the efficacy of cryosurgery in patients with advanced colorectal cancer.

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Flow cytometry of CTCs in the peripheral blood of patients before and after cryotherapy for CRML. (A-D) Analysis of serial dilutions (0.0001%, 0.001%, 0.005% and 0.05%) of human CX-1 tumor cells in normal human blood. (E) Recovery and linear relationship across 3 separate experiments, every separate experiment conducts 10 times (n = 10).
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f0001: Flow cytometry of CTCs in the peripheral blood of patients before and after cryotherapy for CRML. (A-D) Analysis of serial dilutions (0.0001%, 0.001%, 0.005% and 0.05%) of human CX-1 tumor cells in normal human blood. (E) Recovery and linear relationship across 3 separate experiments, every separate experiment conducts 10 times (n = 10).

Mentions: The numbers of CTCs in the peripheral blood of patients before and after cryotherapy for CRML were determined by flow cytometry. A standard curve for the determination of CTCs was generated by adding CX-1 cells to blood obtained from healthy volunteers. Analysis of serial dilutions (0.0001%, 0.001%, 0.005% and 0.05%) of human CX-1 tumor cells in normal human blood demonstrated that the lower detection limit for sensitivity of the method was 0.001%, which is one detected cell per 100,000 white blood cells (Fig. 1A-D). Below this level, background events were unpredictable. Recovery and linearity were highly reproducible according to correlation and regression analysis (Fig. 1E) and the number of tumor cell events recovered correlated positively with the expected number of tumor cell events based on the serial dilutions (R2 = 0.9997).Figure 1.


Analysis of circulating tumor cells in colorectal cancer liver metastasis patients before and after cryosurgery
Flow cytometry of CTCs in the peripheral blood of patients before and after cryotherapy for CRML. (A-D) Analysis of serial dilutions (0.0001%, 0.001%, 0.005% and 0.05%) of human CX-1 tumor cells in normal human blood. (E) Recovery and linear relationship across 3 separate experiments, every separate experiment conducts 10 times (n = 10).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
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getmorefigures.php?uid=PMC5036405&req=5

f0001: Flow cytometry of CTCs in the peripheral blood of patients before and after cryotherapy for CRML. (A-D) Analysis of serial dilutions (0.0001%, 0.001%, 0.005% and 0.05%) of human CX-1 tumor cells in normal human blood. (E) Recovery and linear relationship across 3 separate experiments, every separate experiment conducts 10 times (n = 10).
Mentions: The numbers of CTCs in the peripheral blood of patients before and after cryotherapy for CRML were determined by flow cytometry. A standard curve for the determination of CTCs was generated by adding CX-1 cells to blood obtained from healthy volunteers. Analysis of serial dilutions (0.0001%, 0.001%, 0.005% and 0.05%) of human CX-1 tumor cells in normal human blood demonstrated that the lower detection limit for sensitivity of the method was 0.001%, which is one detected cell per 100,000 white blood cells (Fig. 1A-D). Below this level, background events were unpredictable. Recovery and linearity were highly reproducible according to correlation and regression analysis (Fig. 1E) and the number of tumor cell events recovered correlated positively with the expected number of tumor cell events based on the serial dilutions (R2 = 0.9997).Figure 1.

View Article: PubMed Central - PubMed

ABSTRACT

In this study, we determined the number of peripheral blood circulating tumor cells (CTCs) pre- and post-cryosurgery in patients with colorectal cancer liver metastasis as a reference for understanding the relevance of any changes to the efficacy of cryosurgery. CTC numbers and CTC-related gene expression were measured in the peripheral blood of 55 patients with colorectal liver metastasis at 1 day before and 7 and 30 d after cryoablation using magnetic activated cell sorting (MACS) and fluorescence activated cell sorting (FACS) combined with real-time quantitative PCR (RT-qPCR). The number of CTCs decreased significantly with postoperative time (P < 0.01). Delta cycle threshold values for the CTC-related genes CEA, Ep-CAM, CK18 and CK19 increased significantly after cryoablation. Furthermore, the expression of CEA, Ep-CAM, CK18 and CK19 decreased significantly with time after cryoablation (P < 0.01). RT-qPCR and FACS combined with MACS has significant diagnostic and prognostic value for evaluating the efficacy of cryosurgery in patients with advanced colorectal cancer.

No MeSH data available.


Related in: MedlinePlus