We observed our drug caused a signalling changes in MCF7, which might be relevant to cell invasion. However, MCF7 cells are generally considered as non-invasive, and it seems doesn't invade in matrigel invasion assay. Is there any way to make it invade at the baseline (without my drug treatment)? Some paper suggested it improves its invasion by adding estrogen. Anyone has experience with this? The assay I am currently using is BD matrigel assay. Is there any better alternative assays for MCF7? Any suggestion is welcome.
Dear Esther,
I would actually suggest to consider changing the cell line model to a more invasive one (e.g. MDA-MB-231) in case these cells react the same way to your treatment. Otherwise, there is a report by Weitzenfeld et al. that pre-treatment of the cells with EGF, estogen and TNFa (resembling the TME of luminal tumors) renders them more invasive and aggressive. Alternatively, depending on the genes you see to be changed, you could also consider switching to migration assays instead of invasion (i.e. without the matrigel).
Good luck,
Cindy
You can follow the EMT induction procedures, also we found metastasis in xenograft models with MCF-7 cell along with estrogen therapy.
I think you should include some positive control treatment such as TGFbeta or HGF along with you drug in your experiments. You can treat TGF or HGF treated cells with your drug to check whether the invasive capacity of cells is increased or not?
In our hands regular MCF-7 never invade, but there is at least two papers that can help you. They select the MCF-7 cells by prolonged mammosphere culture, and claim that the cells selected have metastatic potential.
- Guttilla et al. Breast Cancer Res Treat. 2012
- Ziegler et al. Breast Cancer Res Treat. 2014
Good luck!
MCF is a non-invasive line naturally, you can add FBS as a chemoattractant and maybe it will help. We use a method similar to this one to establish standard curves http://csmedia2.corning.com/LifeSciences//media/pdf/an_Chemotaxis_protocol.pdf
And we have both invasive and non invasive, cells in serum free media and cells in low FBS media. The Chemoattractant should be simple enough to try and you won't have to use your drug. I hope this helps
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