死亡受体Fas在TRAIL联合三氧化二砷诱导人胃癌细胞凋亡中的作用  

Effects of Fas in combined treatment of TRAIL and arsenic trioxide-induced apoptosis in human gastric cancer cells

作　　者：段晓秋[1] 王晶[1] 申维喜[1] 杨宇[1] 

机构地区：[1]哈尔滨医科大学附属第二医院肿瘤内科,黑龙江哈尔滨150081

出　　处：《癌变．畸变．突变》2012年第6期422-426,共5页Carcinogenesis,Teratogenesis & Mutagenesis

基　　金：黑龙江省自然科学基金资助项目(D200862)

摘　　要：目的:探讨Fas在肿瘤坏死因子相关凋亡诱导配体(tumor necrosis factor-related apoptosis-inducing ligand,TRAIL)与三氧化二砷(arsenic trioxide,As2O3)联合诱导人胃癌细胞凋亡过程中的作用。方法:采用体外培养人低分化胃腺癌细胞SGC-7901,分别以Fas正义、反义寡核苷酸转染SGC-7901细胞获得Sense、Antisense细胞。以0.5μg/mlAsO+0.2μg/mlTRAIL分别处理未转染细胞、Sense组细胞以及Antisense组细胞,采用MTT法检测24、48、72h后细胞增殖情况;倒置显微镜观察联合作用48h后细胞集落形态;DAPI染色法、流式细胞仪技术检测48h时细胞凋亡率;Westernblot法检测48h细胞中Fas、FADD以及Parp表达情况。结果:与未转染组和Sense组细胞比较,转染Fas反义寡核苷酸后,降低了TRAIL与As2O3联合抑制细胞增殖、促进凋亡的作用(P<0.05),48h时效果最为明显;荧光显微镜下可见死亡细胞明显减少;Westernblot结果显示细胞中Fas、FADD表达降低,Parp活化程度降低(均P<0.05)。结论:Fas在TRAIL与As2O3联合抑制人胃癌细胞增殖、促进细胞凋亡过程中起重要作用。OBJECTIVE：To study the effects of Fas in combined treatment of TRAIL（tumor necrosis factor-related apoptosis-inducing ligand） and arsenic trioxide（As2O3）-induced apoptosis in human gastric cancer SGC-7901 cells. METHODS：SGC-7901 cells were cultured in vitro, Sense and Antisense SGC-7901 cells were obtained from transfection of sense and antisense oligo nucleic acid, respectively. Untransfected, Sense and Antisense cells were treated with 0.5 μg/ml As2O3＋0.2 μg/ml TRAIL, respectively. Cell proliferation was evaluated by MTT assay after 24, 48 and 72 h; cell colony morphologies were examined under inverted microscope after 48 h; apoptosis rate measured by DAPI staining and flow cytometer; and Fas, FADD and Parp expression determined by Western blot. RESULTS：Treatment by Fas antisense oligo nucleic acid transfection decreased the number of dead cells as detected by microscopy, exerted cell growth inhibition and As2O3＋TRAIL-induced apoptosis effects. The expressions of Fas,FADD and activated Parp were all down-regulated （P0.05）. CONCLUSION：Fas signaling pathway may be important in the process of SGC-7901 apoptosis induced by combined treatment of As2O3 and TRAIL.

关 键 词：肿瘤坏死因子相关凋亡诱导配体 三氧化二砷 凋亡 胃癌细胞SGC-7901 FAS 

分 类 号：Q279[生物学—细胞生物学]