I am working with two different liposarcoma cell lines who have a different behaviour when treated with several drugs, I am trying to understand if the p53 status has something to do with that but I did not found any research about, and I don't think that nobody has studied that before.
I am not co-author of Santiago Rello. This is not my field of expertise
Hi Santiago, I work with both cell lines and I don't believe the p53 mutation status of both cell lines have been published before. However, both cell lines were transformed with SV40 Large T antigen during the initial immortalization (http://www.ncbi.nlm.nih.gov/pubmed/1283316), which may affect their p53 function.
Another difference between the cell lines are the variants of the FUS-DDIT3 fusion protein present in each cell line. 402-91 has a Type I, while 1765-92 has a Type VIII fusion protein. Hope that helps.
(**edited because I mistyped 1765-92's fusion variant, it should be Type VIII (8), not type VII (7)!)
In general, when treated with a DNA-damaging agent, such as bleomycin, if p53 levels are increased appreciably and the increased levels are accompanied by increases in the levels of the p53-inducible proteins such as p21, it is likely that the p53 is functional in a cell line. If not, it would be advisable to use antibodies that detect only the mutant p53 in IP and or immunoblotting assays.
I searched a little bit, but I didn't find anything that reported the p53 status of those lines. Unfortunately since those are both myxoid liposarcoma lines, I haven't sequenced them, but I have sequenced p53 in some other liposarcoma cell lines. If you have to know for certain, sequencing them yourself is probably the way to go. I wouldn't recommend using antibodies to mutant p53, those will only be specific for certain mutations, and that's way more work and a lot more expensive than just sequencing.
It won't tell you for certain, but you could also look at the expression levels of p53 in those cell lines compared to a wild-type p53 cell line. Of the three liposarcoma lines I work with, one of them has a p53 mutation, and the other two are wild-type. Because this mutation knocks out the negative feedback loop with Mdm2, levels of p53 protein are higher in the mutant p53 line than they are in the wild-type lines. Additionally, as Divaker pointed out above, you could check the expression levels of p53 target genes, such as p21, PUMA, and Noxa. If they have functional p53, those genes will be expressed more than if they have mutant p53. Again, those won't tell you for certain whether or not there's a p53 mutation, they're not perfect, but they can help point you in the right direction.
I know that my response comes very in late, but how knows? The attached article could be of help.
I did not check the status of the cell lines mentionned in this Q&A.
Best regards
Robert
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