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Identification and Characterization of CXCR4-Positive Gastric Cancer Stem Cells.

Fujita T, Chiwaki F, Takahashi RU, Aoyagi K, Yanagihara K, Nishimura T, Tamaoki M, Komatsu M, Komatsuzaki R, Matsusaki K, Ichikawa H, Sakamoto H, Yamada Y, Fukagawa T, Katai H, Konno H, Ochiya T, Yoshida T, Sasaki H - PLoS ONE (2015)

Bottom Line: In clinical samples, these CXCR4-positive cells were found from not only late metastasis stage (accumulated ascites) but also earlier stage (peritoneal washings).Moreover, treatment with transforming growth factor-β enhanced the anti-cancer effect of docetaxel via induction of cell differentiation/asymmetric cell division of the CXCR4-positive gastric CSCs even in a dormant state.Therefore, differentiation inducers hold promise for obtaining the maximum therapeutic outcome from currently available anti-cancer drugs through re-cycling of CSCs.

View Article: PubMed Central - PubMed

Affiliation: Department of Translational Oncology, National Cancer Center Research Institute, Tokyo, Japan; Second Department of Surgery, Hamamatsu University School of Medicine, Shizuoka, Japan.

ABSTRACT
Diffuse-type solid tumors are often composed of a high proportion of rarely proliferating (i.e., dormant) cancer cells, strongly indicating the involvement of cancer stem cells (CSCs) Although diffuse-type gastric cancer (GC) patients have a poor prognosis due to high-frequent development of peritoneal dissemination (PD), it is limited knowledge that the PD-associated CSCs and efficacy of CSC-targeting therapy in diffuse-type GC. In this study, we established highly metastatic GC cell lines by in vivo selection designed for the enrichment of PD-associated GC cells. By microarray analysis, we found C-X-C chemokine receptor type 4 (CXCR4) can be a novel marker for highly metastatic CSCs, since CXCR4-positive cells can grow anchorage-independently, initiate tumors in mice, be resistant to cytotoxic drug, and produce differentiated daughter cells. In clinical samples, these CXCR4-positive cells were found from not only late metastasis stage (accumulated ascites) but also earlier stage (peritoneal washings). Moreover, treatment with transforming growth factor-β enhanced the anti-cancer effect of docetaxel via induction of cell differentiation/asymmetric cell division of the CXCR4-positive gastric CSCs even in a dormant state. Therefore, differentiation inducers hold promise for obtaining the maximum therapeutic outcome from currently available anti-cancer drugs through re-cycling of CSCs.

No MeSH data available.


Related in: MedlinePlus

CXCR4+ small cells are required for repopulation of all subsets of 60As6 cells.(A) Both CXCR4+ and CXCR4- 60As6 cells were sorted by FACS (left column). The cells of each fraction were cultured for 7 days and then analyzed for the expression levels of CXCR4 (right column). (B) Immunocytochemistry for MUC5AC (green) of a colony derived from a single CXCR4+ small cell. Scale bars represent 50μm. (C) Cell-lineage assay with two different colors. Sorted single CXCR4+ small cells were stained by anti-MUC5AC antibody (red) and CellTracker Green (Invitrogen, green) and for tracing the viable cells for 7 days. Scale bars represent 50μm. (D) RT-PCR of LGR5 in normal gastric epithelial cells (left panel), CXCR4+ small cells and CXCR4- small cells (right panel). Laser microdissection separates the normal gastric mucosa into three regions: pit, neck, and gland.
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pone.0130808.g002: CXCR4+ small cells are required for repopulation of all subsets of 60As6 cells.(A) Both CXCR4+ and CXCR4- 60As6 cells were sorted by FACS (left column). The cells of each fraction were cultured for 7 days and then analyzed for the expression levels of CXCR4 (right column). (B) Immunocytochemistry for MUC5AC (green) of a colony derived from a single CXCR4+ small cell. Scale bars represent 50μm. (C) Cell-lineage assay with two different colors. Sorted single CXCR4+ small cells were stained by anti-MUC5AC antibody (red) and CellTracker Green (Invitrogen, green) and for tracing the viable cells for 7 days. Scale bars represent 50μm. (D) RT-PCR of LGR5 in normal gastric epithelial cells (left panel), CXCR4+ small cells and CXCR4- small cells (right panel). Laser microdissection separates the normal gastric mucosa into three regions: pit, neck, and gland.

Mentions: After sorting the two subpopulation (I and II), we analyze any shift of the cell population (Fig 2A); after 7 days of culture. CXCR4+ small cells corresponding to subpopulation I generated themselves, CXCR4- small cells, and CXCR4- large cells (11%, 62%, and 21%, respectively), whereas CXCR4- small cells corresponding to subpopulation II were able to give rise mainly to themselves (80%) with some large cells (17%) and very few CXCR4+ small cells (2%). These results suggest that CXCR4+ small cells have differentiation potential and produce CXCR4- large cells possibly through a stage of CXCR4- small cells in the scheme of cell differentiation.


Identification and Characterization of CXCR4-Positive Gastric Cancer Stem Cells.

Fujita T, Chiwaki F, Takahashi RU, Aoyagi K, Yanagihara K, Nishimura T, Tamaoki M, Komatsu M, Komatsuzaki R, Matsusaki K, Ichikawa H, Sakamoto H, Yamada Y, Fukagawa T, Katai H, Konno H, Ochiya T, Yoshida T, Sasaki H - PLoS ONE (2015)

CXCR4+ small cells are required for repopulation of all subsets of 60As6 cells.(A) Both CXCR4+ and CXCR4- 60As6 cells were sorted by FACS (left column). The cells of each fraction were cultured for 7 days and then analyzed for the expression levels of CXCR4 (right column). (B) Immunocytochemistry for MUC5AC (green) of a colony derived from a single CXCR4+ small cell. Scale bars represent 50μm. (C) Cell-lineage assay with two different colors. Sorted single CXCR4+ small cells were stained by anti-MUC5AC antibody (red) and CellTracker Green (Invitrogen, green) and for tracing the viable cells for 7 days. Scale bars represent 50μm. (D) RT-PCR of LGR5 in normal gastric epithelial cells (left panel), CXCR4+ small cells and CXCR4- small cells (right panel). Laser microdissection separates the normal gastric mucosa into three regions: pit, neck, and gland.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4481351&req=5

pone.0130808.g002: CXCR4+ small cells are required for repopulation of all subsets of 60As6 cells.(A) Both CXCR4+ and CXCR4- 60As6 cells were sorted by FACS (left column). The cells of each fraction were cultured for 7 days and then analyzed for the expression levels of CXCR4 (right column). (B) Immunocytochemistry for MUC5AC (green) of a colony derived from a single CXCR4+ small cell. Scale bars represent 50μm. (C) Cell-lineage assay with two different colors. Sorted single CXCR4+ small cells were stained by anti-MUC5AC antibody (red) and CellTracker Green (Invitrogen, green) and for tracing the viable cells for 7 days. Scale bars represent 50μm. (D) RT-PCR of LGR5 in normal gastric epithelial cells (left panel), CXCR4+ small cells and CXCR4- small cells (right panel). Laser microdissection separates the normal gastric mucosa into three regions: pit, neck, and gland.
Mentions: After sorting the two subpopulation (I and II), we analyze any shift of the cell population (Fig 2A); after 7 days of culture. CXCR4+ small cells corresponding to subpopulation I generated themselves, CXCR4- small cells, and CXCR4- large cells (11%, 62%, and 21%, respectively), whereas CXCR4- small cells corresponding to subpopulation II were able to give rise mainly to themselves (80%) with some large cells (17%) and very few CXCR4+ small cells (2%). These results suggest that CXCR4+ small cells have differentiation potential and produce CXCR4- large cells possibly through a stage of CXCR4- small cells in the scheme of cell differentiation.

Bottom Line: In clinical samples, these CXCR4-positive cells were found from not only late metastasis stage (accumulated ascites) but also earlier stage (peritoneal washings).Moreover, treatment with transforming growth factor-β enhanced the anti-cancer effect of docetaxel via induction of cell differentiation/asymmetric cell division of the CXCR4-positive gastric CSCs even in a dormant state.Therefore, differentiation inducers hold promise for obtaining the maximum therapeutic outcome from currently available anti-cancer drugs through re-cycling of CSCs.

View Article: PubMed Central - PubMed

Affiliation: Department of Translational Oncology, National Cancer Center Research Institute, Tokyo, Japan; Second Department of Surgery, Hamamatsu University School of Medicine, Shizuoka, Japan.

ABSTRACT
Diffuse-type solid tumors are often composed of a high proportion of rarely proliferating (i.e., dormant) cancer cells, strongly indicating the involvement of cancer stem cells (CSCs) Although diffuse-type gastric cancer (GC) patients have a poor prognosis due to high-frequent development of peritoneal dissemination (PD), it is limited knowledge that the PD-associated CSCs and efficacy of CSC-targeting therapy in diffuse-type GC. In this study, we established highly metastatic GC cell lines by in vivo selection designed for the enrichment of PD-associated GC cells. By microarray analysis, we found C-X-C chemokine receptor type 4 (CXCR4) can be a novel marker for highly metastatic CSCs, since CXCR4-positive cells can grow anchorage-independently, initiate tumors in mice, be resistant to cytotoxic drug, and produce differentiated daughter cells. In clinical samples, these CXCR4-positive cells were found from not only late metastasis stage (accumulated ascites) but also earlier stage (peritoneal washings). Moreover, treatment with transforming growth factor-β enhanced the anti-cancer effect of docetaxel via induction of cell differentiation/asymmetric cell division of the CXCR4-positive gastric CSCs even in a dormant state. Therefore, differentiation inducers hold promise for obtaining the maximum therapeutic outcome from currently available anti-cancer drugs through re-cycling of CSCs.

No MeSH data available.


Related in: MedlinePlus