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Characterization of three cell lines derived from fine needle biopsy of choroidal melanoma with metastatic outcome.

Burgess BL, Rao NP, Eskin A, Nelson SF, McCannel TA - Mol. Vis. (2011)

Bottom Line: In the third, necrotic material from the biopsy prevented further analysis, yet resulted in a stable cell line.Two cell lines had RNA expression profiles similar to the respective primary tumors; the third cell line had a similar RNA expression profile relative to the other two cell lines.FNAB of primary choroidal melanomas resulted in highly characterized, low-passage cell lines, which were stable for the cytogenetic patterns and expression profiles found in the primary tumor.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, University of California, Los Angeles, The Jules Stein Eye Institute, Los Angeles, CA, USA.

ABSTRACT

Purpose: To report three low-passage cell lines from primary choroidal melanoma with metastatic outcome, which were stable for cytogenetic patterns and expression profiles of the primary melanoma.

Methods: In patients with choroidal melanoma, transscleral fine needle aspiration biopsy (FNAB) was performed immediately before plaque placement for (125)iodine brachytherapy or immediately after enucleation. Cells were examined for cytopathology, evaluated by fluorescence in-situ hybridization (FISH) for the centromere of chromosome 3, analyzed by 250K whole genome Mapping Array and U133 plus 2.0 Expression Array, and placed in cell culture. At passage 3, the cell lines were analyzed by Mapping Array and Expression Array.

Results: Three cell lines were propagated from primary choroidal melanomas in three patients who subsequently developed metastasis. Two cell lines were stable for the entire chromosomal aberration pattern of the respective primary tumor. In the third, necrotic material from the biopsy prevented further analysis, yet resulted in a stable cell line. Each cell line had chromosome 3 loss, 6q loss, 8p loss, multiple 8q gain, and 16q loss. Additionally, two cell lines had chromosome 6p gain. Two cell lines had RNA expression profiles similar to the respective primary tumors; the third cell line had a similar RNA expression profile relative to the other two cell lines.

Conclusions: FNAB of primary choroidal melanomas resulted in highly characterized, low-passage cell lines, which were stable for the cytogenetic patterns and expression profiles found in the primary tumor. These cell lines represent novel tools for the study of metastatic choroidal melanoma biology.

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Comparison of Mapping Array results for chromosomal copy number for primary tumor biopsies and their cell lines, where available. Copy number variation analysis, as determined by Affymetrix Genotyping Console, showed identical patterns of aberration between primary tumor biopsies and corresponding cell lines. Gains (green boxes) and losses (red boxes) were largely whole arm in extent. 2± designations signify a total of 4 or more copies of chromosome 8q were detected. All samples had monosomy 3, 6q loss, 8p loss, multiple gains in 8q, and 16q loss. MEL20–06–045 and MEL20–07–070 also had 6p gain.
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f2: Comparison of Mapping Array results for chromosomal copy number for primary tumor biopsies and their cell lines, where available. Copy number variation analysis, as determined by Affymetrix Genotyping Console, showed identical patterns of aberration between primary tumor biopsies and corresponding cell lines. Gains (green boxes) and losses (red boxes) were largely whole arm in extent. 2± designations signify a total of 4 or more copies of chromosome 8q were detected. All samples had monosomy 3, 6q loss, 8p loss, multiple gains in 8q, and 16q loss. MEL20–06–045 and MEL20–07–070 also had 6p gain.

Mentions: Cytogenetically, two of the three cell lines (MEL20–06–039 and MEL20–07–070) demonstrated the chromosomal aberration pattern present in the respective primary melanoma, including monosomy 3, 6q loss, 8p loss, multiple 8q gain and 16q loss (Figure 2). The third cell line (MEL20–06–045), derived from a biopsy that showed necrotic material, had a chromosomal aberration pattern that was nearly identical with the other two cell lines. Additionally, the MEL20–06–045 cell line and both the MEL 20–07–070 cell line and primary tumor exhibited chromosome 6p gain. Moreover, two of the cell lines (MEL20–06–039 and MEL20–07–070) had RNA expression array characteristics similar to the respective primary tumor for the 25 most overexpressed and the 20 most under-expressed genes from a comparative gene list composed of genes most overexpressed and under-expressed in an integrated analysis of choroidal melanoma with chromosome 3 loss versus 6p gain in the absence of chromosome 3 loss (Figure 3A,B) [11]. The third cell line (MEL20–06–045) had a similar profile of gene expression when compared to the two primary melanomas and the other two cell lines (Figure 3C).


Characterization of three cell lines derived from fine needle biopsy of choroidal melanoma with metastatic outcome.

Burgess BL, Rao NP, Eskin A, Nelson SF, McCannel TA - Mol. Vis. (2011)

Comparison of Mapping Array results for chromosomal copy number for primary tumor biopsies and their cell lines, where available. Copy number variation analysis, as determined by Affymetrix Genotyping Console, showed identical patterns of aberration between primary tumor biopsies and corresponding cell lines. Gains (green boxes) and losses (red boxes) were largely whole arm in extent. 2± designations signify a total of 4 or more copies of chromosome 8q were detected. All samples had monosomy 3, 6q loss, 8p loss, multiple gains in 8q, and 16q loss. MEL20–06–045 and MEL20–07–070 also had 6p gain.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3049736&req=5

f2: Comparison of Mapping Array results for chromosomal copy number for primary tumor biopsies and their cell lines, where available. Copy number variation analysis, as determined by Affymetrix Genotyping Console, showed identical patterns of aberration between primary tumor biopsies and corresponding cell lines. Gains (green boxes) and losses (red boxes) were largely whole arm in extent. 2± designations signify a total of 4 or more copies of chromosome 8q were detected. All samples had monosomy 3, 6q loss, 8p loss, multiple gains in 8q, and 16q loss. MEL20–06–045 and MEL20–07–070 also had 6p gain.
Mentions: Cytogenetically, two of the three cell lines (MEL20–06–039 and MEL20–07–070) demonstrated the chromosomal aberration pattern present in the respective primary melanoma, including monosomy 3, 6q loss, 8p loss, multiple 8q gain and 16q loss (Figure 2). The third cell line (MEL20–06–045), derived from a biopsy that showed necrotic material, had a chromosomal aberration pattern that was nearly identical with the other two cell lines. Additionally, the MEL20–06–045 cell line and both the MEL 20–07–070 cell line and primary tumor exhibited chromosome 6p gain. Moreover, two of the cell lines (MEL20–06–039 and MEL20–07–070) had RNA expression array characteristics similar to the respective primary tumor for the 25 most overexpressed and the 20 most under-expressed genes from a comparative gene list composed of genes most overexpressed and under-expressed in an integrated analysis of choroidal melanoma with chromosome 3 loss versus 6p gain in the absence of chromosome 3 loss (Figure 3A,B) [11]. The third cell line (MEL20–06–045) had a similar profile of gene expression when compared to the two primary melanomas and the other two cell lines (Figure 3C).

Bottom Line: In the third, necrotic material from the biopsy prevented further analysis, yet resulted in a stable cell line.Two cell lines had RNA expression profiles similar to the respective primary tumors; the third cell line had a similar RNA expression profile relative to the other two cell lines.FNAB of primary choroidal melanomas resulted in highly characterized, low-passage cell lines, which were stable for the cytogenetic patterns and expression profiles found in the primary tumor.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, University of California, Los Angeles, The Jules Stein Eye Institute, Los Angeles, CA, USA.

ABSTRACT

Purpose: To report three low-passage cell lines from primary choroidal melanoma with metastatic outcome, which were stable for cytogenetic patterns and expression profiles of the primary melanoma.

Methods: In patients with choroidal melanoma, transscleral fine needle aspiration biopsy (FNAB) was performed immediately before plaque placement for (125)iodine brachytherapy or immediately after enucleation. Cells were examined for cytopathology, evaluated by fluorescence in-situ hybridization (FISH) for the centromere of chromosome 3, analyzed by 250K whole genome Mapping Array and U133 plus 2.0 Expression Array, and placed in cell culture. At passage 3, the cell lines were analyzed by Mapping Array and Expression Array.

Results: Three cell lines were propagated from primary choroidal melanomas in three patients who subsequently developed metastasis. Two cell lines were stable for the entire chromosomal aberration pattern of the respective primary tumor. In the third, necrotic material from the biopsy prevented further analysis, yet resulted in a stable cell line. Each cell line had chromosome 3 loss, 6q loss, 8p loss, multiple 8q gain, and 16q loss. Additionally, two cell lines had chromosome 6p gain. Two cell lines had RNA expression profiles similar to the respective primary tumors; the third cell line had a similar RNA expression profile relative to the other two cell lines.

Conclusions: FNAB of primary choroidal melanomas resulted in highly characterized, low-passage cell lines, which were stable for the cytogenetic patterns and expression profiles found in the primary tumor. These cell lines represent novel tools for the study of metastatic choroidal melanoma biology.

Show MeSH
Related in: MedlinePlus