Hello,
I have done my masters and the results observed was that nanoencapsulated siRNA had antiproliferative activity (MTT assay) and that mock or naked siRNA did not (silencing by qPCR was 65%). Confocal images showed that there was more siRNA-Cy5 in cytoplasm if encapsulated, compared to free administration of siRNA-Cy5. These experiments was with brand new bought MCF7 cell line cultured with RPMI and 20% of FBS.
Now I am in another country, working with MCF7 cell line that was available. Here they culture in DMEM and 10% of FBS and free siRNA is reducing proliferation by 20-30% (did not do qPCR yet). Other than that, confocal images shows a lot of siRNA both encapsulated or not.
I dont know if the FBS amount can interfere so much in this, but it is a hypothesis. But also, in theory, free siRNA shouldn´t go inside cells.
Maybe the cells are different in some way, and are more sensible? I dont know.
The siRNA was designed and ordered with exactly the same properties and at the same company.
Other than that, mock nanoparticle is also showing some toxicity. So maybe is a cell problem?
Can anyone help me to see some light?
Hello Ana Beatriz Valle ,
I'd also suspect the cell line. Try to get the cells from your old lab to compare. If you see the same changes in response you can go and look for other reasons.
You may find this interesting:
Article Genetic and transcriptional evolution alters cancer cell lin...
Of course, it's also very easy to increase the FBS and see what happens...
Hope that helps
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