Medknow Publications on behalf of the Neurological Society of India
Abstract
Purpose: To detect the expressions of Fas/DcR3 and to investigate the
cytotoxic effects of RGD-FasL on pituitary adenoma cells. Materials and
Methods: Fas/DcR3 mRNAs were detected by Reverse transcription
polymerase chain reaction (RT-PCR) and their surface expressions were
measured by flow cytometry. Cytotoxicities exerted by FasL and
newly-constructed RGD-FasL on tumor cells were measured with
3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT)
assay. The apoptotic cells were examined by electron microscopy and the
induced apoptosis was determined by agarose gel electrophoresis. The
cell cycle was assessed by flow cytometry with ANNEXIN V FITC/PI. The
expressions of caspases, Bcl-2, RANKL and JNK2 were detected by Western
blotting. Results: Fas/DcR3 was expressed in GH3/MMQ/AtT20 cells. The
cytotoxic effects of RGD-FasL on tumor cells were seen in a
dose-dependent manner. These cells showed the same sensitivity to
RGD-FasL as to FasL. RGD-FasL induced apoptosis and G1/G0 arrest. The
expressions of caspase-8/9/3, RANKL, JNK2 were increased while that of
Bcl-2 was decreased with treatment of RGD-FasL. Conclusions: Fas can be
a novel target for the treatment of pituitary adenomas. RGD-FasL
induces apoptosis of pituitary adenoma cells through caspase
activation