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Retaining the long-survive capacity of Circulating Tumor Cells (CTCs) followed by xeno-transplantation: not only from metastatic cancer of the breast but also of prostate cancer patients.

Rossi E, Rugge M, Facchinetti A, Pizzi M, Nardo G, Barbieri V, Manicone M, De Faveri S, Chiara Scaini M, Basso U, Amadori A, Zamarchi R - Oncoscience (2013)

Bottom Line: Human CTCs were found in 8 out of 8 murine peripheral blood (muPB) and in 6 out of 8 murine bone marrow (muBM) samples, after a median follow-up of 10.3 months.Our assay showed higher successful rate than those previously reported in breast cancer and hepatocellular carcinoma.This is the first experimental evidence that as few as 50 EpCAM-positive prostate cancer CTCs putatively contain metastasis-initiating-cells (MIC).

View Article: PubMed Central - PubMed

Affiliation: Department of Surgery, Oncology and Gastroenterology, Oncology Section, University of Padova, Padova, Italy ; IOV-IRCCS, Padova, Italy.

ABSTRACT
We investigated whether Circulating Tumor Cells (CTCs) isolated from epithelial tumors could survive and grow in xenotransplants. To this purpose, EpCAM-positive CTCs were enriched by CellSearch platform the only FDA-cleared automated platform that quantifies tumor burden in peripheral blood and provides clinical evidence of predictive and prognostic value. The CTCs were isolated from metastatic prostate (n=6) and breast (n=2) cancer patients. The xenograft assay was developed in 8-week-old NOD/SCID mice that were subcutaneously injected with increasing amounts of CTCs (ranging from 50 to 3000). Human CTCs were found in 8 out of 8 murine peripheral blood (muPB) and in 6 out of 8 murine bone marrow (muBM) samples, after a median follow-up of 10.3 months. Six out of 8 spleens were positive for human cytokeratin. Our assay showed higher successful rate than those previously reported in breast cancer and hepatocellular carcinoma. The role of EpCAM during carcinogenesis is controversial. The identification of human CTCs in muPB, muBM and spleen demonstrates that the EpCAM-positive fraction of CTCs retains the migratory capacity. This is the first experimental evidence that as few as 50 EpCAM-positive prostate cancer CTCs putatively contain metastasis-initiating-cells (MIC).

No MeSH data available.


Related in: MedlinePlus

A) Human CTCs in peripheral blood (PB) and DTCs in bone marrow (BM) of xeno-transplanted mice. The gallery shows the same cell stained for the combination (Comp) of huCK PE (green) and DAPI (violet); huCK PE; DAPI; muCD45 APC and background control. Red squares indicate cells classified as CTCs/DTCs.Line 1 and 2. CellSearch analysis of some rare human CTCs in murine PB and DTCs in BM samples of a xenograft assay from prostate cancer (xenog #1).Line 3. CellSearch analysis of CTCs in murine PB sample of a xenograft assay from metastatic breast cancer (MBC) (xenog #3) , (Analyzer II, CellSearch).Single CTCs, exhibiting regular CK staining were detected in the PB samples of xenografts #1 and #3, whereas a small neoplastic embolus (with 3 clearly distinguished DAPI+ nuclei) was detected in the BM of xenograft #1.B) Neoplastic cells within the spleen of xeno-transplanted mice. The picture shows haematoxylin and eosin (H&E) staining (left panel) and pan-cytokeratin immunostaining (right panel) of murine spleens from xenografts #1 and #3 and of a mice control (original magnification, 20X).xenog #1: small neoplastic emboli found in perisplenic small vessels;xenog #3: pan-cytokeratin immunostaining disclosed small groups of anti-human cytokeratin positive cells (undetectable at the H&E staining), into the spleen red pulp;ctrl: the spleen from a control mouse does not display any morphological (left picture) or immunohystochemical (right picture) evidence of anti-human cytokeratin positive cells. One out of 6 representative samples is shown.
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Figure 3: A) Human CTCs in peripheral blood (PB) and DTCs in bone marrow (BM) of xeno-transplanted mice. The gallery shows the same cell stained for the combination (Comp) of huCK PE (green) and DAPI (violet); huCK PE; DAPI; muCD45 APC and background control. Red squares indicate cells classified as CTCs/DTCs.Line 1 and 2. CellSearch analysis of some rare human CTCs in murine PB and DTCs in BM samples of a xenograft assay from prostate cancer (xenog #1).Line 3. CellSearch analysis of CTCs in murine PB sample of a xenograft assay from metastatic breast cancer (MBC) (xenog #3) , (Analyzer II, CellSearch).Single CTCs, exhibiting regular CK staining were detected in the PB samples of xenografts #1 and #3, whereas a small neoplastic embolus (with 3 clearly distinguished DAPI+ nuclei) was detected in the BM of xenograft #1.B) Neoplastic cells within the spleen of xeno-transplanted mice. The picture shows haematoxylin and eosin (H&E) staining (left panel) and pan-cytokeratin immunostaining (right panel) of murine spleens from xenografts #1 and #3 and of a mice control (original magnification, 20X).xenog #1: small neoplastic emboli found in perisplenic small vessels;xenog #3: pan-cytokeratin immunostaining disclosed small groups of anti-human cytokeratin positive cells (undetectable at the H&E staining), into the spleen red pulp;ctrl: the spleen from a control mouse does not display any morphological (left picture) or immunohystochemical (right picture) evidence of anti-human cytokeratin positive cells. One out of 6 representative samples is shown.

Mentions: Figure 3-A illustrates an example of human CTCs and DTCs recovered from muPB and muBM of two representative xenografts, as shown by the Analyzer II device (CellSearch). As indicated in Table 1 (xenograft's data; in supplementary file), human CTCs were found in 8 out of 8 muPB samples; the mean CTC number was 11.4 ± 12.6 (median 8, range 2-42 CTCs/ 0.75 ml muPB). Moreover, human DTCs were identified in 6 out of 8 muBM samples; the mean DTC number was 1.8 ± 0.8 (median 2, range 1-3 DTCs/ 0.75 ml muBM).


Retaining the long-survive capacity of Circulating Tumor Cells (CTCs) followed by xeno-transplantation: not only from metastatic cancer of the breast but also of prostate cancer patients.

Rossi E, Rugge M, Facchinetti A, Pizzi M, Nardo G, Barbieri V, Manicone M, De Faveri S, Chiara Scaini M, Basso U, Amadori A, Zamarchi R - Oncoscience (2013)

A) Human CTCs in peripheral blood (PB) and DTCs in bone marrow (BM) of xeno-transplanted mice. The gallery shows the same cell stained for the combination (Comp) of huCK PE (green) and DAPI (violet); huCK PE; DAPI; muCD45 APC and background control. Red squares indicate cells classified as CTCs/DTCs.Line 1 and 2. CellSearch analysis of some rare human CTCs in murine PB and DTCs in BM samples of a xenograft assay from prostate cancer (xenog #1).Line 3. CellSearch analysis of CTCs in murine PB sample of a xenograft assay from metastatic breast cancer (MBC) (xenog #3) , (Analyzer II, CellSearch).Single CTCs, exhibiting regular CK staining were detected in the PB samples of xenografts #1 and #3, whereas a small neoplastic embolus (with 3 clearly distinguished DAPI+ nuclei) was detected in the BM of xenograft #1.B) Neoplastic cells within the spleen of xeno-transplanted mice. The picture shows haematoxylin and eosin (H&E) staining (left panel) and pan-cytokeratin immunostaining (right panel) of murine spleens from xenografts #1 and #3 and of a mice control (original magnification, 20X).xenog #1: small neoplastic emboli found in perisplenic small vessels;xenog #3: pan-cytokeratin immunostaining disclosed small groups of anti-human cytokeratin positive cells (undetectable at the H&E staining), into the spleen red pulp;ctrl: the spleen from a control mouse does not display any morphological (left picture) or immunohystochemical (right picture) evidence of anti-human cytokeratin positive cells. One out of 6 representative samples is shown.
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Related In: Results  -  Collection

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Figure 3: A) Human CTCs in peripheral blood (PB) and DTCs in bone marrow (BM) of xeno-transplanted mice. The gallery shows the same cell stained for the combination (Comp) of huCK PE (green) and DAPI (violet); huCK PE; DAPI; muCD45 APC and background control. Red squares indicate cells classified as CTCs/DTCs.Line 1 and 2. CellSearch analysis of some rare human CTCs in murine PB and DTCs in BM samples of a xenograft assay from prostate cancer (xenog #1).Line 3. CellSearch analysis of CTCs in murine PB sample of a xenograft assay from metastatic breast cancer (MBC) (xenog #3) , (Analyzer II, CellSearch).Single CTCs, exhibiting regular CK staining were detected in the PB samples of xenografts #1 and #3, whereas a small neoplastic embolus (with 3 clearly distinguished DAPI+ nuclei) was detected in the BM of xenograft #1.B) Neoplastic cells within the spleen of xeno-transplanted mice. The picture shows haematoxylin and eosin (H&E) staining (left panel) and pan-cytokeratin immunostaining (right panel) of murine spleens from xenografts #1 and #3 and of a mice control (original magnification, 20X).xenog #1: small neoplastic emboli found in perisplenic small vessels;xenog #3: pan-cytokeratin immunostaining disclosed small groups of anti-human cytokeratin positive cells (undetectable at the H&E staining), into the spleen red pulp;ctrl: the spleen from a control mouse does not display any morphological (left picture) or immunohystochemical (right picture) evidence of anti-human cytokeratin positive cells. One out of 6 representative samples is shown.
Mentions: Figure 3-A illustrates an example of human CTCs and DTCs recovered from muPB and muBM of two representative xenografts, as shown by the Analyzer II device (CellSearch). As indicated in Table 1 (xenograft's data; in supplementary file), human CTCs were found in 8 out of 8 muPB samples; the mean CTC number was 11.4 ± 12.6 (median 8, range 2-42 CTCs/ 0.75 ml muPB). Moreover, human DTCs were identified in 6 out of 8 muBM samples; the mean DTC number was 1.8 ± 0.8 (median 2, range 1-3 DTCs/ 0.75 ml muBM).

Bottom Line: Human CTCs were found in 8 out of 8 murine peripheral blood (muPB) and in 6 out of 8 murine bone marrow (muBM) samples, after a median follow-up of 10.3 months.Our assay showed higher successful rate than those previously reported in breast cancer and hepatocellular carcinoma.This is the first experimental evidence that as few as 50 EpCAM-positive prostate cancer CTCs putatively contain metastasis-initiating-cells (MIC).

View Article: PubMed Central - PubMed

Affiliation: Department of Surgery, Oncology and Gastroenterology, Oncology Section, University of Padova, Padova, Italy ; IOV-IRCCS, Padova, Italy.

ABSTRACT
We investigated whether Circulating Tumor Cells (CTCs) isolated from epithelial tumors could survive and grow in xenotransplants. To this purpose, EpCAM-positive CTCs were enriched by CellSearch platform the only FDA-cleared automated platform that quantifies tumor burden in peripheral blood and provides clinical evidence of predictive and prognostic value. The CTCs were isolated from metastatic prostate (n=6) and breast (n=2) cancer patients. The xenograft assay was developed in 8-week-old NOD/SCID mice that were subcutaneously injected with increasing amounts of CTCs (ranging from 50 to 3000). Human CTCs were found in 8 out of 8 murine peripheral blood (muPB) and in 6 out of 8 murine bone marrow (muBM) samples, after a median follow-up of 10.3 months. Six out of 8 spleens were positive for human cytokeratin. Our assay showed higher successful rate than those previously reported in breast cancer and hepatocellular carcinoma. The role of EpCAM during carcinogenesis is controversial. The identification of human CTCs in muPB, muBM and spleen demonstrates that the EpCAM-positive fraction of CTCs retains the migratory capacity. This is the first experimental evidence that as few as 50 EpCAM-positive prostate cancer CTCs putatively contain metastasis-initiating-cells (MIC).

No MeSH data available.


Related in: MedlinePlus