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Precancerous stem cells have the potential for both benign and malignant differentiation.

Chen L, Shen R, Ye Y, Pu XA, Liu X, Duan W, Wen J, Zimmerer J, Wang Y, Liu Y, Lasky LC, Heerema NA, Perrotti D, Ozato K, Kuramochi-Miyagawa S, Nakano T, Yates AJ, Carson WE, Lin H, Barsky SH, Gao JX - PLoS ONE (2007)

Bottom Line: However, their precursors-namely, precancerous stem cells (pCSCs) -have not been characterized.Mechanistically, the pCSCs are regulated by the PIWI/AGO family gene called piwil2.Our results provide clear evidence that a single clone of pCSCs has the potential for both benign and malignant differentiation, depending on the environmental cues.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Ohio State University Medical Center, Columbus, Ohio, United States of America.

ABSTRACT
Cancer stem cells (CSCs) have been identified in hematopoietic and solid tumors. However, their precursors-namely, precancerous stem cells (pCSCs) -have not been characterized. Here we experimentally define the pCSCs that have the potential for both benign and malignant differentiation, depending on environmental cues. While clonal pCSCs can develop into various types of tissue cells in immunocompetent mice without developing into cancer, they often develop, however, into leukemic or solid cancers composed of various types of cancer cells in immunodeficient mice. The progress of the pCSCs to cancers is associated with the up-regulation of c-kit and Sca-1, as well as with lineage markers. Mechanistically, the pCSCs are regulated by the PIWI/AGO family gene called piwil2. Our results provide clear evidence that a single clone of pCSCs has the potential for both benign and malignant differentiation, depending on the environmental cues. We anticipate pCSCs to be a novel target for the early detection, prevention, and therapy of cancers.

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pCSCs have long-term repopulating activity.Congenic CD45.1 B6 mice were lethally irradiated and injected i.v. with 0.5∼1×106 2C4, 3B5C or 3B6C cells along with 2∼5×105 recipient-type BM cells. Donor-specific CD45.2+ lymphoid (CD3ε+) and myeloid (CD11b+ or Gr-1+) cells were monitored by flow cytometric analysis of blood cells starting from 4 wks post transfer, once every two wks, until 18 wks (A). The mice were sacrificed 10 moths post transfer, and the blood and BM cells were collected for HANDS-Nested DNA PCR to identify donor-derived cells (B). To verify the self-renewal capability of the long-term repopulated donor cells, 1×106 BM cells from the primary recipients were injected i.v. into the secondary recipients, which were sacrificed 10 wks post transfer. The pCSC-derived neor gene in the BM, liver, and spleen was determined by HANDS-Nested DNA PCR (C). The data shown in B are from a recipient with transient expansion of pCSC-derived hematopoietic cells at 8 and 13 wks post transfer, and the data shown in C & D are from one of 3 experiments (5∼10 mice/group/expt).
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pone-0000293-g002: pCSCs have long-term repopulating activity.Congenic CD45.1 B6 mice were lethally irradiated and injected i.v. with 0.5∼1×106 2C4, 3B5C or 3B6C cells along with 2∼5×105 recipient-type BM cells. Donor-specific CD45.2+ lymphoid (CD3ε+) and myeloid (CD11b+ or Gr-1+) cells were monitored by flow cytometric analysis of blood cells starting from 4 wks post transfer, once every two wks, until 18 wks (A). The mice were sacrificed 10 moths post transfer, and the blood and BM cells were collected for HANDS-Nested DNA PCR to identify donor-derived cells (B). To verify the self-renewal capability of the long-term repopulated donor cells, 1×106 BM cells from the primary recipients were injected i.v. into the secondary recipients, which were sacrificed 10 wks post transfer. The pCSC-derived neor gene in the BM, liver, and spleen was determined by HANDS-Nested DNA PCR (C). The data shown in B are from a recipient with transient expansion of pCSC-derived hematopoietic cells at 8 and 13 wks post transfer, and the data shown in C & D are from one of 3 experiments (5∼10 mice/group/expt).

Mentions: The aborted in vitro hematopoietic differentiation may indicate how strict environmental cues influence pCSC differentiation and/or survival. To test this hypothesis, a competitive in vivo repopulating assay was performed [19]. Lethally irradiated CD45.1+ congenic B6 mice were injected i.v. with both 2C4, 3B5C or 3B6C cells and recipient-type bone marrow (BM) cells. The latter, BM cells were added to the injection to minimize recipient deaths due to lethal irradiation. Donor-derived CD45.2+ lymphoid (CD3ε+) and myeloid (CD11b+ or Gr-1+) cells in the peripheral blood were monitored by flow cytometry beginning ∼4 wks post-transplant. CD45.2+ donor cells were not significantly detected until ∼8 wks after transfer (Fig. 2a). About 0.5∼10% more CD45.2+CD11b+ and CD45.2+Gr-1+ cells were detected, depending on individual mice or experiments (5∼10 mice/expt) (Fig. 2a). Two weeks later, the pCSC-derived CD45.2+ cells decreased unexpectedly to less than 0.5% in the peripheral blood and were gone at ∼13 wks (Fig. 2a). These results suggest that, like our experiments performed in vitro, the pCSCs also have multipotent but incomplete differentiation in vivo, eventually leading to cell apoptosis. The incomplete differentiation may explain the low myeloid engraftment, as well as the absence of lymphoid engraftment. We define the process as differentiation-induced cell death (DICD), probably a protective mechanism that prevents pCSCs from progressing to malignant cells.


Precancerous stem cells have the potential for both benign and malignant differentiation.

Chen L, Shen R, Ye Y, Pu XA, Liu X, Duan W, Wen J, Zimmerer J, Wang Y, Liu Y, Lasky LC, Heerema NA, Perrotti D, Ozato K, Kuramochi-Miyagawa S, Nakano T, Yates AJ, Carson WE, Lin H, Barsky SH, Gao JX - PLoS ONE (2007)

pCSCs have long-term repopulating activity.Congenic CD45.1 B6 mice were lethally irradiated and injected i.v. with 0.5∼1×106 2C4, 3B5C or 3B6C cells along with 2∼5×105 recipient-type BM cells. Donor-specific CD45.2+ lymphoid (CD3ε+) and myeloid (CD11b+ or Gr-1+) cells were monitored by flow cytometric analysis of blood cells starting from 4 wks post transfer, once every two wks, until 18 wks (A). The mice were sacrificed 10 moths post transfer, and the blood and BM cells were collected for HANDS-Nested DNA PCR to identify donor-derived cells (B). To verify the self-renewal capability of the long-term repopulated donor cells, 1×106 BM cells from the primary recipients were injected i.v. into the secondary recipients, which were sacrificed 10 wks post transfer. The pCSC-derived neor gene in the BM, liver, and spleen was determined by HANDS-Nested DNA PCR (C). The data shown in B are from a recipient with transient expansion of pCSC-derived hematopoietic cells at 8 and 13 wks post transfer, and the data shown in C & D are from one of 3 experiments (5∼10 mice/group/expt).
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC1808425&req=5

pone-0000293-g002: pCSCs have long-term repopulating activity.Congenic CD45.1 B6 mice were lethally irradiated and injected i.v. with 0.5∼1×106 2C4, 3B5C or 3B6C cells along with 2∼5×105 recipient-type BM cells. Donor-specific CD45.2+ lymphoid (CD3ε+) and myeloid (CD11b+ or Gr-1+) cells were monitored by flow cytometric analysis of blood cells starting from 4 wks post transfer, once every two wks, until 18 wks (A). The mice were sacrificed 10 moths post transfer, and the blood and BM cells were collected for HANDS-Nested DNA PCR to identify donor-derived cells (B). To verify the self-renewal capability of the long-term repopulated donor cells, 1×106 BM cells from the primary recipients were injected i.v. into the secondary recipients, which were sacrificed 10 wks post transfer. The pCSC-derived neor gene in the BM, liver, and spleen was determined by HANDS-Nested DNA PCR (C). The data shown in B are from a recipient with transient expansion of pCSC-derived hematopoietic cells at 8 and 13 wks post transfer, and the data shown in C & D are from one of 3 experiments (5∼10 mice/group/expt).
Mentions: The aborted in vitro hematopoietic differentiation may indicate how strict environmental cues influence pCSC differentiation and/or survival. To test this hypothesis, a competitive in vivo repopulating assay was performed [19]. Lethally irradiated CD45.1+ congenic B6 mice were injected i.v. with both 2C4, 3B5C or 3B6C cells and recipient-type bone marrow (BM) cells. The latter, BM cells were added to the injection to minimize recipient deaths due to lethal irradiation. Donor-derived CD45.2+ lymphoid (CD3ε+) and myeloid (CD11b+ or Gr-1+) cells in the peripheral blood were monitored by flow cytometry beginning ∼4 wks post-transplant. CD45.2+ donor cells were not significantly detected until ∼8 wks after transfer (Fig. 2a). About 0.5∼10% more CD45.2+CD11b+ and CD45.2+Gr-1+ cells were detected, depending on individual mice or experiments (5∼10 mice/expt) (Fig. 2a). Two weeks later, the pCSC-derived CD45.2+ cells decreased unexpectedly to less than 0.5% in the peripheral blood and were gone at ∼13 wks (Fig. 2a). These results suggest that, like our experiments performed in vitro, the pCSCs also have multipotent but incomplete differentiation in vivo, eventually leading to cell apoptosis. The incomplete differentiation may explain the low myeloid engraftment, as well as the absence of lymphoid engraftment. We define the process as differentiation-induced cell death (DICD), probably a protective mechanism that prevents pCSCs from progressing to malignant cells.

Bottom Line: However, their precursors-namely, precancerous stem cells (pCSCs) -have not been characterized.Mechanistically, the pCSCs are regulated by the PIWI/AGO family gene called piwil2.Our results provide clear evidence that a single clone of pCSCs has the potential for both benign and malignant differentiation, depending on the environmental cues.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Ohio State University Medical Center, Columbus, Ohio, United States of America.

ABSTRACT
Cancer stem cells (CSCs) have been identified in hematopoietic and solid tumors. However, their precursors-namely, precancerous stem cells (pCSCs) -have not been characterized. Here we experimentally define the pCSCs that have the potential for both benign and malignant differentiation, depending on environmental cues. While clonal pCSCs can develop into various types of tissue cells in immunocompetent mice without developing into cancer, they often develop, however, into leukemic or solid cancers composed of various types of cancer cells in immunodeficient mice. The progress of the pCSCs to cancers is associated with the up-regulation of c-kit and Sca-1, as well as with lineage markers. Mechanistically, the pCSCs are regulated by the PIWI/AGO family gene called piwil2. Our results provide clear evidence that a single clone of pCSCs has the potential for both benign and malignant differentiation, depending on the environmental cues. We anticipate pCSCs to be a novel target for the early detection, prevention, and therapy of cancers.

Show MeSH
Related in: MedlinePlus