If I were to say, \"A CAF ( cancer-associated fibroblast ) can cause greater mig

Question

If I were to say, "A CAF (cancer-associated fibroblast) can cause greater migration of breast tumor cells," what experimental approach would you use to prove this hypothesis? Include:

a) the experimental approach.

b) a general outline of the procedure for that approach, i.e. an overview of how the protocol is conducted, including critical reagents or steps.

c) expected results, including any specific marker proteins associated with the assay that would change or vary.

Nothing too lengthy, please and thank you!

Explanation / Answer

By research, it had been observed and indicated by researchers that the cancer associated fibroblasts have a capability of migration, especially the ones associated near the desmoplastic areas. To prove the same:

1) Experimental approach

The cells will be cultured in the medium along with their stimulation and then motility would be checked along with the characterisation of the accelerated motility configuration.

2) Experimental setup

To characterise the same, quantitative proteomics (SILAC) require to be performed . Cells with breast cancer cells are cultured in conditioned media for stimulation and in serum free media as well and the stimulation of the cells were checked.

Cell proliferation assay was done using crystal violet assay. Micro environment of the cells are also impor

3) Markers associated with breast cancer associated fibroblasts

Though, there are no specific markers for the breast CAFs, there have been observed high expressions for -SMA, p53, podoplanin, CD10, fibroblast activation protein (FAP), matrix metalloproteinases (MMPs), tenascin-C and platelet-derived growth factor (PDGFR/). It was observed that theses cells promote proliferation of breast cancer by secretion of various cytokines and various growth factors such as FGFs, HGF, TGF- and SDF-1 .

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