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KIT and CD34 relative expression were comparable among the IGF1Rlow wild-type GISTs of group 3 and the wild-type GISTs of group 4. Thus, relatively high KIT and CD34 expression distinguished the IGF1Rhigh group 4 wild-type GISTs from the IGF1Rhigh group 5, and this high KIT and CD34 expression was shared with the wild-type IGF1Rlow group 3 GISTs.
The only responder in this trial had very high KIT protein expression, supporting the hypothesis that c-KIT aberrant melanomas are responsive to c-KIT inhibitors such as Gleevec.
The high Kit level in SCF-cultured cells was reminiscent of a mechanism involving SCF as a modulator of its own receptor expression as described for IL-3, CSF-1 and GM-CSF [25].
FISH analysis was successful in 10 out of 12 tumours showing high KIT expression.
When tumours with low and high KIT expression were analysed as one group (positive KIT expression), the difference was statistically significant (P=0.0414).
The adaptor-ligated cDNA fragments were released from the beads via digestion by EcoP15I (NEB), end-repaired by KOD DNA Polymerase (Blunting High Kit, Toyobo), and subsequently ligated to a second barcoding adaptor using the T4 Ligase reagent provided with the Ligation high Ver. 2 Kit (Toyobo).
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The highest yield of DNA was obtained by the FA-SS kit, which was more than two times that of the second highest kit (MB-PS).
Plasma adiponectin and insulin concentrations were determined using ELISA kits (Human Adiponectin ELISA, High Sensitivity Kit, BioVendor-Labaratorni Medicine for adiponectin; and Insulin, Canine ELISA, Mercodia AB, Uppsala, Sweden for insulin).
For determination of rDNA repeat copy number, genomic DNA was extracted by Dr. GenTLE® (from yeast) High Recovery Kit (Takara Bio Inc., Shiga, Japan).
All products were evaluated on the Agilent Bioanalyzer using the High Sensitivity kit.
A high sensitivity kit was used for analysing G-CSF.
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