Chen Lukui, Tian Xinhua, Li Wenzhu, Agarwal Abhishek, Zhuang Guohong
Department of Neurosurgery, Zhongshan Hospital, Xiamen University, Xiamen, Fujian - 361 004, China.
Neurol India. 2009 Jan-Feb;57(1):28-30. doi: 10.4103/0028-3886.48808.
To detect the expressions of Fas/DcR3 and to investigate the cytotoxic effects of RGD-FasL on pituitary adenoma cells.
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.
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.
Fas can be a novel target for the treatment of pituitary adenomas. RGD-FasL induces apoptosis of pituitary adenoma cells through caspase activation.
检测Fas/DcR3的表达，并研究RGD-FasL对垂体腺瘤细胞的细胞毒性作用。
采用逆转录聚合酶链反应（RT-PCR）检测Fas/DcR3 mRNA，通过流式细胞术检测其表面表达。用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）法检测FasL和新构建的RGD-FasL对肿瘤细胞的细胞毒性。通过电子显微镜检查凋亡细胞，用琼脂糖凝胶电泳测定诱导凋亡情况。采用流式细胞术结合膜联蛋白V FITC/PI评估细胞周期。通过蛋白质免疫印迹法检测半胱天冬酶、Bcl-2、核因子κB受体活化因子配体（RANKL）和应激活化蛋白激酶2（JNK2）的表达。
Fas/DcR3在GH3/MMQ/AtT20细胞中表达。RGD-FasL对肿瘤细胞的细胞毒性呈剂量依赖性。这些细胞对RGD-FasL和FasL表现出相同的敏感性。RGD-FasL诱导细胞凋亡和G1/G0期阻滞。用RGD-FasL处理后，半胱天冬酶-8/9/3、RANKL、JNK2的表达增加，而Bcl-2的表达降低。
Fas可能是治疗垂体腺瘤的新靶点。RGD-FasL通过激活半胱天冬酶诱导垂体腺瘤细胞凋亡。
