探讨肿瘤坏死因子相关凋亡诱导配体(tumor necrosis factor related apoptosis inducing ligand，TRAIL)对人骨巨细胞瘤(giant cell tumor of bone ,GCT)的作用及干扰素 γ(interferon γ,IFN γ)对TRAIL抗瘤活性的影响,并探讨其影响机制。方法： 选取2003年-2006年于301医院骨科手术切除的骨巨细胞瘤新鲜标本9例，将取自标本的骨巨细胞瘤细胞分为:对照组, TRAIL组，IFN γ组, IFN γ诱导后加入TRAIL组,应用CCK 8法检测TRAIL、IFN γ单独及联合作用对骨巨细胞瘤细胞的增殖抑制作用，流式细胞仪、TUNEL法分析IFN γ对TRAIL凋亡诱导作用的影响，RT PCR法检测IFN γ作用前后肿瘤细胞DR4、DR5、TRAIL的表达。结果:骨巨细胞瘤细胞对照组存活率为（98.64±0.31）%，TRAIL (100、 500、1 000 μg/L)单独作用对肿瘤细胞增殖无明显抑制作用;IFN γ(500、1 000 U /ml)单独作用后,细胞的存活率分别为(94.05±1.89)%、(9047±2.66)%。IFN γ(500 U /ml)诱导24 h后与TRAIL(100、200 μg/L)联合作用,细胞的存活率分别为(84.65±246)%、(77.65±3.14)%。两者联合作用后流式细胞仪测得联合作用组凋亡率为(27.94±2.88)%、(38.65±2.46) %，对照组凋亡率为(1.77±0.49)%；TUNEL法测得联合作用组凋亡率分別为(19.63±3.51)%、(29.28±4.80)%，对照组的凋亡率为(1.1±0.17)%。RT PCR 结果显示，IFN γ作用24 h后肿瘤细胞TRAIL、DR4、DR5 mRNA表达明显上调。结论：骨巨细胞瘤细胞对TRAIL不敏感, IFN γ可以明显提高TRAIL诱导骨巨细胞瘤细胞凋亡的敏感性,其机制可能与IFN γ上调TRAIL、DR4、DR5mRNA的表达有关。

Abstract Objective: To explore the anti tumor effect of tumor necrosis factor related apoptosis inducing ligand (TRAIL) against giant cell tumor of bone (GCT), and to study the influence of IFN γ on the anti tumor effects of TRAIL and the possible mechanisms. MethodsFresh specimens were obtained from 9 patients who were pathologically diagnosed as having GCT in the orthopaedics department of No. 301 Hospital PLA from 2003 to 2006. The GCT cells from specimens were divided into 4 groups: control group, TRAIL group, IFN γ group and IFN γ plus TRAIL group. The cell inhibition was examined by CCK 8 assay. Cell apoptosis was detected by flow cytometric analysis and TUNEL assays. RT PCR was applied to semi quantitatively assay the mRNA expression of TRAIL, DR4 and DR5 in GCT cells before and after the treatment with IFN γ (500 U/ml). Results: The GCT cell survival rate was （98.64±0.31）% in the control group. TRAIL alone (100, 500,1 000 μg/L) had no obvious effect on tumor cell proliferation. After treatment with IFN γ at 500 and 1 000 U/ml , the tumor cell survival rates were (94.05±1.89) % and (90.47±2.66) %, respectively. After treatement with IFN γ (500 U /ml) and TRAIL (100,200 μg/L), the tumor cell survival rates were (84.65±246)% and (77.65±3.14)%, respectively; and FCM showed that their apoptosis rates were (2794±288)% and (38.65±2.46)%, respectively ; the apoptosis rate was (1.77±0.49)% in the control group. TUNEL revealed that the apoptosis rates were (1963±351)% and (29.28±4.80)% , respectively; and the rate was (1.1±0.17)% in the control group. RT PCR showed that the expression levels of TRAIL, DR4 and DR5 in tumor cells were up regulated after treatment with IFN γ (500 U /ml) for 24 h. Conclusion: GCT cells are not sensitive to TRAIL. IFN γ can up regulate the apoptosis rate of GCT cells induced by TRAIL, which might be associated with the up regulation of TRAIL, DR4 and DR5 mRNA expression by IFN γ.

国家高技术研究发展（863）计划资助项目（No.2002AA214081）