PMID:22363644

Citation	Justiniano, SE, Mathew, A, Mitra, S, Manivannan, SN and Simcox, A (2012) Loss of the tumor suppressor Pten promotes proliferation of Drosophila melanogaster cells in vitro and gives rise to continuous cell lines. PLoS ONE 7:e31417
Abstract	In vivo analysis of Drosophila melanogaster has enhanced our understanding of many biological processes, notably the mechanisms of heredity and development. While in vivo analysis of mutants has been a strength of the field, analyzing fly cells in culture is valuable for cell biological, biochemical and whole genome approaches in which large numbers of homogeneous cells are required. An efficient genetic method to derive Drosophila cell lines using expression of an oncogenic form of Ras (Ras(V12)) has been developed. Mutations in tumor suppressors, which are known to cause cell hyperproliferation in vivo, could provide another method for generating Drosophila cell lines. Here we screened Drosophila tumor suppressor mutations to test if they promoted cell proliferation in vitro. We generated primary cultures and determined when patches of proliferating cells first emerged. These cells emerged on average at 37 days in wild-type cultures. Using this assay we found that a Pten mutation had a strong effect. Patches of proliferating cells appeared on average at 11 days and the cultures became confluent in about 3 weeks, which is similar to the timeframe for cultures expressing Ras(V12). Three Pten mutant cell lines were generated and these have now been cultured for between 250 and 630 cell doublings suggesting the life of the mutant cells is likely to be indefinite. We conclude that the use of Pten mutants is a powerful means to derive new Drosophila cell lines.
Links	PubMed PMC3283623 Online version:10.1371/journal.pone.0031417
Keywords	Animals; Biological Assay; Cell Line; Cell Proliferation; Drosophila Proteins/deficiency; Drosophila Proteins/metabolism; Drosophila melanogaster/cytology; Drosophila melanogaster/enzymology; Hyperplasia; Mutant Proteins/metabolism; Mutation/genetics; Neoplasms/metabolism; Neoplasms/pathology; PTEN Phosphohydrolase/deficiency; PTEN Phosphohydrolase/metabolism; Time Factors; Tumor Suppressor Proteins/metabolism; ras Proteins/metabolism
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
DROME:Q9U470		GO:0008283: cell proliferation	ECO:0000314:		P		Figure 1 shows cell proliferation in pten mutants to be within a statistically significant time as the positive control. Figure 2 shows the similarity in pten mutants compared to oncogenic cultures known for cell proliferation.	complete CACAO 4457

References

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