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ALDH1-high ovarian cancer stem-like cells can be isolated from serous and clear cell adenocarcinoma cells, and ALDH1 high expression is associated with poor prognosis.

Kuroda T, Hirohashi Y, Torigoe T, Yasuda K, Takahashi A, Asanuma H, Morita R, Mariya T, Asano T, Mizuuchi M, Saito T, Sato N - PLoS ONE (2013)

Bottom Line: We isolated ovarian CSCs/CICs as an aldehyde dehydrogenase 1 high (ALDH1(high)) population from 6 EOC cell lines (3 serous adenocarcinomas and 3 clear cell adenocarcinomas) by the ALDEFLUOR assay.ALDH1(high) cells could also be isolated from 8 of 11 primary ovarian carcinoma samples.Immunohistochemical staining revealed that higher ALDH1 expression levels in ovary cancer cases are related to poorer prognosis in both serous adenocarcinoma cases and clear cell adenocarcinoma cases.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Sapporo Medical University School of Medicine, Chuo-Ku, Sapporo, Japan.

ABSTRACT
Cancer stem-like cells (CSCs)/cancer-initiating cells (CICs) are defined as a small population of cancer cells that have high tumorigenicity. Furthermore, CSCs/CICs are resistant to several cancer therapies, and CSCs/CICs are therefore thought to be responsible for cancer recurrence after treatment and distant metastasis. In epithelial ovarian cancer (EOC) cases, disease recurrence after chemotherapy is frequently observed, suggesting ovarian CSCs/CICs are involved. There are four major histological subtypes in EOC, and serous adenocarcinoma and clear cell adenocarcinoma are high-grade malignancies. We therefore analyzed ovarian CSCs/CICs from ovarian carcinoma cell lines (serous adenocarcinoma and clear cell adenocarcinoma) and primary ovarian cancer cells in this study. We isolated ovarian CSCs/CICs as an aldehyde dehydrogenase 1 high (ALDH1(high)) population from 6 EOC cell lines (3 serous adenocarcinomas and 3 clear cell adenocarcinomas) by the ALDEFLUOR assay. ALDH1(high) cells showed greater sphere-forming ability, higher tumorigenicity and greater invasive capability, indicating that ovarian CSCs/CICs are enriched in ALDH1(high) cells. ALDH1(high) cells could also be isolated from 8 of 11 primary ovarian carcinoma samples. Immunohistochemical staining revealed that higher ALDH1 expression levels in ovary cancer cases are related to poorer prognosis in both serous adenocarcinoma cases and clear cell adenocarcinoma cases. Taken together, the results indicate that ALDH1 is a marker for ovarian CSCs/CICs and that the expression level of ALDH1 might be a novel biomarker for prediction of poor prognosis.

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Comparison of sphere-forming ability and invasion capability of ALDH1high cells and ALDH1low cells.A Sphere-forming assay One thousand ALDH1high cells and ALDH1low cells were cultured in a floating condition. The spheres were counted on day 10. Magnification of images: ×100. Criterion for spheroid size: over 100µm. Each value is the mean number of spheroids± SD. *P values. B Single cell sphere-forming assay Sorted ALDH1high cells and ALDH1low cells were cultured in a floating condition at a single cell per well. The spheres were counted on day 12. Only ALDH1high cells from AMOC-s and ES-2 cells initiated single cell spheroids. Magnification of images: ×200. Each values is the mean percentage of spheroids ± SD. C Matrigel invasion assay Images: Matrigel-invading cells derived from ALDH1high/low cells of ES-2 and TOV-21G cells. Magnification of images: ×100. Each values is the mean number of invading cells ± SD. *P values.
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pone-0065158-g002: Comparison of sphere-forming ability and invasion capability of ALDH1high cells and ALDH1low cells.A Sphere-forming assay One thousand ALDH1high cells and ALDH1low cells were cultured in a floating condition. The spheres were counted on day 10. Magnification of images: ×100. Criterion for spheroid size: over 100µm. Each value is the mean number of spheroids± SD. *P values. B Single cell sphere-forming assay Sorted ALDH1high cells and ALDH1low cells were cultured in a floating condition at a single cell per well. The spheres were counted on day 12. Only ALDH1high cells from AMOC-s and ES-2 cells initiated single cell spheroids. Magnification of images: ×200. Each values is the mean percentage of spheroids ± SD. C Matrigel invasion assay Images: Matrigel-invading cells derived from ALDH1high/low cells of ES-2 and TOV-21G cells. Magnification of images: ×100. Each values is the mean number of invading cells ± SD. *P values.

Mentions: Since CSCs/CICs are known to form spheres in floating culture conditions [29], we analyzed ALDH1high cells by a sphere-forming assay. A total of 103 cells per well were sorted and incubated into a 6–well plate in an anchorage-independent environment. At day 10, ALDH1high cells derived from AMOC-2 cells showed greater sphere-forming ability than that of ALDH1low cells. Similar results were obtained from ES-2 cells, RMG-1 cells and TOV-21G cells (Figure 2A). Since the sphere-forming assay is not suitable for quantification of the ratios of CSCs/CICs in ALDH1high cells, we performed a single cell sphere-forming assay. Sphere formation was observed in 4.86% of the wells of ALDH1high cells derived from AMOC-2 cells and in 3.13% of the wells of ALDH1high cells derived from ES-2 cells (Figure 2B). On the other hand, wells of ALDH1low cells derived from both AMOC-2 and ES-2 cells did not show any sphere formation.


ALDH1-high ovarian cancer stem-like cells can be isolated from serous and clear cell adenocarcinoma cells, and ALDH1 high expression is associated with poor prognosis.

Kuroda T, Hirohashi Y, Torigoe T, Yasuda K, Takahashi A, Asanuma H, Morita R, Mariya T, Asano T, Mizuuchi M, Saito T, Sato N - PLoS ONE (2013)

Comparison of sphere-forming ability and invasion capability of ALDH1high cells and ALDH1low cells.A Sphere-forming assay One thousand ALDH1high cells and ALDH1low cells were cultured in a floating condition. The spheres were counted on day 10. Magnification of images: ×100. Criterion for spheroid size: over 100µm. Each value is the mean number of spheroids± SD. *P values. B Single cell sphere-forming assay Sorted ALDH1high cells and ALDH1low cells were cultured in a floating condition at a single cell per well. The spheres were counted on day 12. Only ALDH1high cells from AMOC-s and ES-2 cells initiated single cell spheroids. Magnification of images: ×200. Each values is the mean percentage of spheroids ± SD. C Matrigel invasion assay Images: Matrigel-invading cells derived from ALDH1high/low cells of ES-2 and TOV-21G cells. Magnification of images: ×100. Each values is the mean number of invading cells ± SD. *P values.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3675199&req=5

pone-0065158-g002: Comparison of sphere-forming ability and invasion capability of ALDH1high cells and ALDH1low cells.A Sphere-forming assay One thousand ALDH1high cells and ALDH1low cells were cultured in a floating condition. The spheres were counted on day 10. Magnification of images: ×100. Criterion for spheroid size: over 100µm. Each value is the mean number of spheroids± SD. *P values. B Single cell sphere-forming assay Sorted ALDH1high cells and ALDH1low cells were cultured in a floating condition at a single cell per well. The spheres were counted on day 12. Only ALDH1high cells from AMOC-s and ES-2 cells initiated single cell spheroids. Magnification of images: ×200. Each values is the mean percentage of spheroids ± SD. C Matrigel invasion assay Images: Matrigel-invading cells derived from ALDH1high/low cells of ES-2 and TOV-21G cells. Magnification of images: ×100. Each values is the mean number of invading cells ± SD. *P values.
Mentions: Since CSCs/CICs are known to form spheres in floating culture conditions [29], we analyzed ALDH1high cells by a sphere-forming assay. A total of 103 cells per well were sorted and incubated into a 6–well plate in an anchorage-independent environment. At day 10, ALDH1high cells derived from AMOC-2 cells showed greater sphere-forming ability than that of ALDH1low cells. Similar results were obtained from ES-2 cells, RMG-1 cells and TOV-21G cells (Figure 2A). Since the sphere-forming assay is not suitable for quantification of the ratios of CSCs/CICs in ALDH1high cells, we performed a single cell sphere-forming assay. Sphere formation was observed in 4.86% of the wells of ALDH1high cells derived from AMOC-2 cells and in 3.13% of the wells of ALDH1high cells derived from ES-2 cells (Figure 2B). On the other hand, wells of ALDH1low cells derived from both AMOC-2 and ES-2 cells did not show any sphere formation.

Bottom Line: We isolated ovarian CSCs/CICs as an aldehyde dehydrogenase 1 high (ALDH1(high)) population from 6 EOC cell lines (3 serous adenocarcinomas and 3 clear cell adenocarcinomas) by the ALDEFLUOR assay.ALDH1(high) cells could also be isolated from 8 of 11 primary ovarian carcinoma samples.Immunohistochemical staining revealed that higher ALDH1 expression levels in ovary cancer cases are related to poorer prognosis in both serous adenocarcinoma cases and clear cell adenocarcinoma cases.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Sapporo Medical University School of Medicine, Chuo-Ku, Sapporo, Japan.

ABSTRACT
Cancer stem-like cells (CSCs)/cancer-initiating cells (CICs) are defined as a small population of cancer cells that have high tumorigenicity. Furthermore, CSCs/CICs are resistant to several cancer therapies, and CSCs/CICs are therefore thought to be responsible for cancer recurrence after treatment and distant metastasis. In epithelial ovarian cancer (EOC) cases, disease recurrence after chemotherapy is frequently observed, suggesting ovarian CSCs/CICs are involved. There are four major histological subtypes in EOC, and serous adenocarcinoma and clear cell adenocarcinoma are high-grade malignancies. We therefore analyzed ovarian CSCs/CICs from ovarian carcinoma cell lines (serous adenocarcinoma and clear cell adenocarcinoma) and primary ovarian cancer cells in this study. We isolated ovarian CSCs/CICs as an aldehyde dehydrogenase 1 high (ALDH1(high)) population from 6 EOC cell lines (3 serous adenocarcinomas and 3 clear cell adenocarcinomas) by the ALDEFLUOR assay. ALDH1(high) cells showed greater sphere-forming ability, higher tumorigenicity and greater invasive capability, indicating that ovarian CSCs/CICs are enriched in ALDH1(high) cells. ALDH1(high) cells could also be isolated from 8 of 11 primary ovarian carcinoma samples. Immunohistochemical staining revealed that higher ALDH1 expression levels in ovary cancer cases are related to poorer prognosis in both serous adenocarcinoma cases and clear cell adenocarcinoma cases. Taken together, the results indicate that ALDH1 is a marker for ovarian CSCs/CICs and that the expression level of ALDH1 might be a novel biomarker for prediction of poor prognosis.

Show MeSH
Related in: MedlinePlus