I am not convinced that anoikis is a major way through which GBM cells die (see the Lefranc et al. attached article).

Validated protocols for cell death analyses are here attached (Galluzzi et al.).

You will encounter technical problems in trying to recapitulate actual anoikis in GBM cells because i) GBM cells rarely invade adjacent tissues (including brain parenchyma) through vessels and ii) they diffusely invade the brain parenchyma.

You can try to use an anti-integrin antibody in 2D GBM cell cultures to induce "mechanic" anoikis and then you should "simply" applied apoptosis-related protocols.

However, you should first determine what is/are the major extracellular components secreted by GBM cells during their "substratum attachment" on the plstic bottom of the flask (or even glass). The ECM secreted components vary from one GBM cell line to another one (see Belot et al.).

You could also use "anti-ion channel" compounds. For this point, I advice you to look carefully at the abundant publications by Harald Sontheimer on this subject.

A very nice article has also been recently posted by Luis Pardo on Kv10.1 with respect to GBM cells.

I hope it could help you.

Robet

Dear Robert Kiss,

Thank You for your valuable comments and suggestions.

Regards

Ashutosh

There are several factors of various interrelated pathways belongs to the Anoikis but the AKT protein is mainly responsible for the Anoikis (detachment following death of cell). thus qRT-PCR and western blotting can be used for the quantification of akt gene regulation and its protein product measurement and by which you can assess the Anoikis. eventually, i am suggesting a outcome related to anoikis on the basis of my knowledge, i.e. anoikis is triggered by down-regulation of akt gene leads to pose a lower level of its protein inside the cell.