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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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Related in: MedlinePlus

Phase contrast photomicrographs of cell lines in subconfluent and overgrowth conditions. At passage 3, cell lines MEL20–06–039 (A and B), MEL20–06–045 (C and D) and MEL20–07–070 (E and F) show early stage propagation (A, C, and E) and late stage confluent propagation with heaped up cells and no evidence of contact inhibition (B, D, and F). Melanin production is evident in two cell lines (MEL20–06–045 and MEL20–07–070). Original magnification (A, C, E) 200×, (B, D, F) 100×. Scale bar is 30 µm.
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f1: Phase contrast photomicrographs of cell lines in subconfluent and overgrowth conditions. At passage 3, cell lines MEL20–06–039 (A and B), MEL20–06–045 (C and D) and MEL20–07–070 (E and F) show early stage propagation (A, C, and E) and late stage confluent propagation with heaped up cells and no evidence of contact inhibition (B, D, and F). Melanin production is evident in two cell lines (MEL20–06–045 and MEL20–07–070). Original magnification (A, C, E) 200×, (B, D, F) 100×. Scale bar is 30 µm.

Mentions: The three characterized cell lines with monosomy 3 were from patients with choroidal melanoma who developed clinical evidence of melanoma metastasis <1.5 years after primary melanoma diagnosis and treatment; all three patients subsequently died from liver failure due to metastatic disease. All three cell lines demonstrated stable propagation through at least passage 6 over a period of 2.5 years. Two of the three cell lines persisted in melanin production beyond passage 6 and none of the cultures exhibited contact inhibition in over-growth conditions (Figure 1).


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)

Phase contrast photomicrographs of cell lines in subconfluent and overgrowth conditions. At passage 3, cell lines MEL20–06–039 (A and B), MEL20–06–045 (C and D) and MEL20–07–070 (E and F) show early stage propagation (A, C, and E) and late stage confluent propagation with heaped up cells and no evidence of contact inhibition (B, D, and F). Melanin production is evident in two cell lines (MEL20–06–045 and MEL20–07–070). Original magnification (A, C, E) 200×, (B, D, F) 100×. Scale bar is 30 µm.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f1: Phase contrast photomicrographs of cell lines in subconfluent and overgrowth conditions. At passage 3, cell lines MEL20–06–039 (A and B), MEL20–06–045 (C and D) and MEL20–07–070 (E and F) show early stage propagation (A, C, and E) and late stage confluent propagation with heaped up cells and no evidence of contact inhibition (B, D, and F). Melanin production is evident in two cell lines (MEL20–06–045 and MEL20–07–070). Original magnification (A, C, E) 200×, (B, D, F) 100×. Scale bar is 30 µm.
Mentions: The three characterized cell lines with monosomy 3 were from patients with choroidal melanoma who developed clinical evidence of melanoma metastasis <1.5 years after primary melanoma diagnosis and treatment; all three patients subsequently died from liver failure due to metastatic disease. All three cell lines demonstrated stable propagation through at least passage 6 over a period of 2.5 years. Two of the three cell lines persisted in melanin production beyond passage 6 and none of the cultures exhibited contact inhibition in over-growth conditions (Figure 1).

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