Reversion of monochromosome-mediated suppression of tumorigenicity in malignant melanoma by retroviral transduction

Yan A. Su, Mike E. Ray, Ti Lin, Nancy E. Seidel, David M. Bodine, Paul S. Meltzer, Jeffrey M. Trent

Research output: Contribution to journalArticle

Abstract

We have developed a general strategy to reverse monochromosome suppression of the malignant phenotypes by retroviral transduction. Our approach involved the introduction of a retroviral expression vector-carried cDNA library into a chromosome 6-suppressed melanoma subline UACC-903(+6) [J. M. Trent et al., Science (Washington DC), 247: 568-571, 1990]. The cDNA library was constructed from polyadenylated RNA isolated from the suppressed UACC-903(+6) cells, packaged into high-titer amphotropic retrovirus particles, and transduced into UACC-903(+6) cells. Revertant his(R) transductants were selected by isolating colony-forming cells in soft agar. A total of 121 large (< 150 μm) colonies was picked from soft agar culture with 18 of 121 (15%) established as permanent sublines. The revertant sublines demonstrated 7-58% cloning efficiency upon plating in agar, in contrast to <0.05% for the UACC-90(+6) subline. All 18 revertant sublines, termed SRS1-SRS18 (for 'selection of revertants for suppression'), displayed a reduced population-doubling time, with 9 of 18 showing focus formation in monolayer similar to the parental (nonsuppressed) cell line. Preliminary evidence for reversion of the suppressed phenotype by injection of cells into athymic nude mice has been completed for one revertant subline. Southern analysis has demonstrated integration of the retroviral vector sequence in all 18 sublines. This approach should facilitate the identification of genes involved in the tumorigenic phenotype of malignant melanoma, and is readily adaptable to other model systems.

Original languageEnglish (US)
Pages (from-to)3186-3191
Number of pages6
JournalCancer Research
Volume56
Issue number14
StatePublished - Jul 15 1996

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Fingerprint Dive into the research topics of 'Reversion of monochromosome-mediated suppression of tumorigenicity in malignant melanoma by retroviral transduction'. Together they form a unique fingerprint.

  • Cite this

    Su, Y. A., Ray, M. E., Lin, T., Seidel, N. E., Bodine, D. M., Meltzer, P. S., & Trent, J. M. (1996). Reversion of monochromosome-mediated suppression of tumorigenicity in malignant melanoma by retroviral transduction. Cancer Research, 56(14), 3186-3191.