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Assessment of cancer cell line representativeness using microarrays for Merkel cell carcinoma.

Daily K, Coxon A, Williams JS, Lee CC, Coit DG, Busam KJ, Brownell I - J. Invest. Dermatol. (2014)

Bottom Line: When compared with publicly available cancer lines, WaGa and Mkl-1 cells were similar to other neuroendocrine tumors, but the variant cell lines were not.Spectral karyotyping and short tandem repeat analysis of the UISO cells matched the original cell line's description, ruling out contamination.Our results validate the use of transcriptome analysis to assess the cancer cell line representativeness and indicate that UISO, MCC13, and MCC26 cell lines are not representative of MCC tumors, whereas WaGa and Mkl-1 more closely model MCC.

View Article: PubMed Central - PubMed

Affiliation: Dermatology Branch, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, USA.

ABSTRACT
When using cell lines to study cancer, phenotypic similarity to the original tumor is paramount. Yet, little has been done to characterize how closely Merkel cell carcinoma (MCC) cell lines model native tumors. To determine their similarity to MCC tumor samples, we characterized MCC cell lines via gene expression microarrays. Using whole transcriptome gene expression signatures and a computational bioinformatic approach, we identified significant differences between variant cell lines (UISO, MCC13, and MCC26) and fresh frozen MCC tumors. Conversely, the classic WaGa and Mkl-1 cell lines more closely represented the global transcriptome of MCC tumors. When compared with publicly available cancer lines, WaGa and Mkl-1 cells were similar to other neuroendocrine tumors, but the variant cell lines were not. WaGa and Mkl-1 cells grown as xenografts in mice had histological and immunophenotypical features consistent with MCC, whereas UISO xenograft tumors were atypical for MCC. Spectral karyotyping and short tandem repeat analysis of the UISO cells matched the original cell line's description, ruling out contamination. Our results validate the use of transcriptome analysis to assess the cancer cell line representativeness and indicate that UISO, MCC13, and MCC26 cell lines are not representative of MCC tumors, whereas WaGa and Mkl-1 more closely model MCC.

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UISO xenograft tumors are histologically atypical for MCCRepresentative images of hematoxylin and eosin staining (H&E) and immunohistochemical staining of WaGa and UISO xenograft tumors in NOD/Scid mice. Scale bar = 100μm.
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Figure 4: UISO xenograft tumors are histologically atypical for MCCRepresentative images of hematoxylin and eosin staining (H&E) and immunohistochemical staining of WaGa and UISO xenograft tumors in NOD/Scid mice. Scale bar = 100μm.

Mentions: To confirm that the distinct expression profile observed in cultured UISO cells correlates with an atypical tumor phenotype in vivo, we grew xenograft tumors with UISO, WaGa, and Mkl-1 cells in immunocompromised mice. Histologically, WaGa tumors showed features consistent with MCC: sheets of tumor cells with scant cytoplasm, rounded nuclei, inconspicuous nucleoli, stippled chromatin patterning, and a brisk mitotic rate (Figure 4). WaGa xenograft tissue also resembled the immunophenotype of typical MCC tumors with positive immunostainng for the neuroendocrine markers chromogranin, NCAM1 (CD56), and ENO2. The MCC marker KRT20 and pan-cytokeratin (AE1/AE3) both stained with a characteristic paranuclear dot pattern in WaGa tumors, and the SCLC marker TTF1 was negative. As a MCV positive cell line (Houben et al., 2010), WaGa xenografts stained with CM2B4, an anti-MCV T antigen antibody. Mkl-1 tumors showed immunohistochemical features similar to WaGa tumors, consistent with typical MCC tumors (data not shown).


Assessment of cancer cell line representativeness using microarrays for Merkel cell carcinoma.

Daily K, Coxon A, Williams JS, Lee CC, Coit DG, Busam KJ, Brownell I - J. Invest. Dermatol. (2014)

UISO xenograft tumors are histologically atypical for MCCRepresentative images of hematoxylin and eosin staining (H&E) and immunohistochemical staining of WaGa and UISO xenograft tumors in NOD/Scid mice. Scale bar = 100μm.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4366303&req=5

Figure 4: UISO xenograft tumors are histologically atypical for MCCRepresentative images of hematoxylin and eosin staining (H&E) and immunohistochemical staining of WaGa and UISO xenograft tumors in NOD/Scid mice. Scale bar = 100μm.
Mentions: To confirm that the distinct expression profile observed in cultured UISO cells correlates with an atypical tumor phenotype in vivo, we grew xenograft tumors with UISO, WaGa, and Mkl-1 cells in immunocompromised mice. Histologically, WaGa tumors showed features consistent with MCC: sheets of tumor cells with scant cytoplasm, rounded nuclei, inconspicuous nucleoli, stippled chromatin patterning, and a brisk mitotic rate (Figure 4). WaGa xenograft tissue also resembled the immunophenotype of typical MCC tumors with positive immunostainng for the neuroendocrine markers chromogranin, NCAM1 (CD56), and ENO2. The MCC marker KRT20 and pan-cytokeratin (AE1/AE3) both stained with a characteristic paranuclear dot pattern in WaGa tumors, and the SCLC marker TTF1 was negative. As a MCV positive cell line (Houben et al., 2010), WaGa xenografts stained with CM2B4, an anti-MCV T antigen antibody. Mkl-1 tumors showed immunohistochemical features similar to WaGa tumors, consistent with typical MCC tumors (data not shown).

Bottom Line: When compared with publicly available cancer lines, WaGa and Mkl-1 cells were similar to other neuroendocrine tumors, but the variant cell lines were not.Spectral karyotyping and short tandem repeat analysis of the UISO cells matched the original cell line's description, ruling out contamination.Our results validate the use of transcriptome analysis to assess the cancer cell line representativeness and indicate that UISO, MCC13, and MCC26 cell lines are not representative of MCC tumors, whereas WaGa and Mkl-1 more closely model MCC.

View Article: PubMed Central - PubMed

Affiliation: Dermatology Branch, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, USA.

ABSTRACT
When using cell lines to study cancer, phenotypic similarity to the original tumor is paramount. Yet, little has been done to characterize how closely Merkel cell carcinoma (MCC) cell lines model native tumors. To determine their similarity to MCC tumor samples, we characterized MCC cell lines via gene expression microarrays. Using whole transcriptome gene expression signatures and a computational bioinformatic approach, we identified significant differences between variant cell lines (UISO, MCC13, and MCC26) and fresh frozen MCC tumors. Conversely, the classic WaGa and Mkl-1 cell lines more closely represented the global transcriptome of MCC tumors. When compared with publicly available cancer lines, WaGa and Mkl-1 cells were similar to other neuroendocrine tumors, but the variant cell lines were not. WaGa and Mkl-1 cells grown as xenografts in mice had histological and immunophenotypical features consistent with MCC, whereas UISO xenograft tumors were atypical for MCC. Spectral karyotyping and short tandem repeat analysis of the UISO cells matched the original cell line's description, ruling out contamination. Our results validate the use of transcriptome analysis to assess the cancer cell line representativeness and indicate that UISO, MCC13, and MCC26 cell lines are not representative of MCC tumors, whereas WaGa and Mkl-1 more closely model MCC.

Show MeSH
Related in: MedlinePlus