Anat Cell Biol.  2015 Mar;48(1):44-53. 10.5115/acb.2015.48.1.44.

Glioblastoma specific antigens, GD2 and CD90, are not involved in cancer stemness

• Woo SR
• Oh YT
• An JY
• Kang BG
• Nam DH
• Joo KM

• KMID: 2046074
• DOI: http://doi.org/10.5115/acb.2015.48.1.44

Abstract

Glioblastoma multiforme (GBM) is the most malignant World Health Organization grade IV brain tumor. GBM patients have a poor prognosis because of its resistance to standard therapies, such as chemotherapy and radiation. Since stem-like cells have been associated with the treatment resistance of GBM, novel therapies targeting the cancer stem cell (CSC) population is critically required. However, GBM CSCs share molecular and functional characteristics with normal neural stem cells (NSCs). To elucidate differential therapeutic targets of GBM CSCs, we compared surface markers of GBM CSCs with adult human NSCs and found that GD2 and CD90 were specifically overexpressed in GBM CSCs. We further tested whether the GBM CSC specific markers are associated with the cancer stemness using primarily cultured patient-derived GBM cells. However, results consistently indicated that GBM cells with or without GD2 and CD90 had similar in vitro sphere formation capacity, a functional characteristics of CSCs. Therefore, GD2 and CD90, GBM specific surface markers, might not be used as specific therapeutic targets for GBM CSCs, although they could have other clinical utilities.

Keyword

Glioblastoma multiforme; Cancer stem cell; Therapeutic marker; GD2; CD90

MeSH Terms

Adult
Brain Neoplasms
Drug Therapy
Glioblastoma*
Humans
Neoplastic Stem Cells
Neural Stem Cells
Prognosis
World Health Organization

Figure

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  Fig. 1 Sox2 and nestin were highly regulated in both normal neural stem cells (NSCs) and glioblastoma multiforme (GBM) cells. 464T and 532T GBM cells, as well as 682 and 779 normal NSCs were stained by anti-Sox2 (upper, green) and anti-nestin (lower, green) antibodies for immunocytochemistry. Nuclei of all cells were stained by DAPI (blue).

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  Fig. 2 Glioblastoma multiforme cells unlike neural stem cells generated tumor mass in orthotopic brain xenograft models. Hematoxylin and eosin (H&E) and immunohistochemistry staining for proliferating cell nuclear antigen (PCNA) expression were performed in representative whole brains of the orthotopic xenograft. Bar chart summary of average staining intensity of PCNA in 3 randomly selected hot spot regions (*P<0.05).

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  Fig. 3 Cell surface markers of 242 kinds were analyzed in normal neural stem cells (NSCs) and glioblastoma multiforme (GBM) cells. 464T (B) and 532T (D) GBM cells, as well as 682 (A) and 779 (C) normal NSCs were stained by BD Lyoplate Human Cell Surface Marker Screening Panel containg 242 antibodies. All cells were counted by flow cytometer BD Caliber & LSR II. Data were analyzed by BD FACSDive software.

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  Fig. 4 Positive expression of GD2 and CD90 were significantly increased in glioblastoma multiforme (GBM) cells than neural stem cells (NSCs). BD Lyoplate Human Cell Surface Marker Screening showing high regulated markers of >50% were selected in GBM cells, and analyzed with the expressed changes in NSCs. Data was expressed by the fold changes of each value in GBM cells as compared with NSCs for the selected 11 markers.

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  Fig. 5 GBM cells with high levels of GD2 or CD90 protein do not have characteristics of cancer stemness. 464T cells were stained by anti-GD2 (A) and anti-CD90 (B) antibodies. The high-regulated and low-regulated cells of GD2 or CD90, were sorted by fluorescence activated cell sorting analysis. The sorted cells and parent cells were tested for the ability of neurosphere formation according to the expressed levels of GD2 (C) or CD90 (D) by the limiting dilution assay.

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