In GM-CSF Restrain Mitochondria Pathway Apoptosis NF-kB Is Not the Only Way.

After stimulated by GM-CSF, Akt and IkB kinase (IKK) can be phosphorylated by activated PIK3-kinase. Activated IKK phosphorylate IkB, making NF-kB released and translocate to nucleus. Granulocyte-macrophage colony-stimulating factor (GM-CSF) restrain TF-1 cell mitochondria pathway apoptosis through PI3K pathway, so we asked if NF-κB is the only way in this process.

All group were plused TF-1 cells 2.5X10⁵/250μl, group A plused hGM-CSF (100ng/ml ) 50μl; group B plused hGM-CSF (100ng/ml) 50 μl and SN50 (50 μmol/L) 20μl; group C plused hGM-CSF 50μl and SN50 200μl. After cultured for 48 hours, all groups were dyed with Annexin and PI, analyzed in FCM. At the same time all groups were dyed by diluted MitoCapture to analyze mitochondria membrane electrical potential. Drop experimental group B 10μl on slips, so as group C, then observe it under Fluorescence microscopes FL1 channel.

Group A cells that have normal mitochondria membrane electrical potential can form poly-MitoCapture in it, and inspired red fluorescence. Group C cells that have monomer MitoCapture was inspired green fluorescence. The red fluorescence intensity of control, group A, B, C means is 368.93, 1137.59, 1033.61 and 618.37 respectively. Group A and B retained a statistically significant difference with control (P<0.01). Group C retained a statistically significant difference with control (P<0.01), group A and group B (P<0.01). This investigation proves that GM-CSF restrain TF-1 cells can be incompletely blocked by SN50, which is the specificity inhibitor of NF-kB. So we get the conclusion that NF-kB is an important but not the only pathway in the process of GM-CSF restrain TF-1 cells apoptosis. It also suggests that there maybe some other pathway in this process.