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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

段晓秋/王 晶/申维喜/杨 宇*   

  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
  • 收稿日期:2012-04-16 修回日期:2012-09-02 出版日期:2011-11-30 发布日期:2011-11-30
  • 通讯作者: 杨宇,E-mail:yangy@medmail.com.cn
  • 作者简介:段晓秋(1983- ),女,山东省临沂市人,硕士研究生,研究方向:肿瘤的化学预防。
  • 基金资助:

    黑龙江省自然科学基金资助项目(D200862)

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

DUAN Xiao-qiu,WANG Jing,SHEN Wei-xi,YANG Yu*   

  1. Department of Medical Oncology, the Second Affiliated of Harbin Medical University 150081, Heilongjiang, China
  • Received:2012-04-16 Revised:2012-09-02 Online:2011-11-30 Published:2011-11-30
  • Contact: YANG Yu,E-mail:yangy@medmail.com.cn
  • About author:段晓秋(1983- ),女,山东省临沂市人,硕士研究生,研究方向:肿瘤的化学预防。
  • Supported by:

    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0.5 μg/ml As2O3+0.2 μg/ml TRAIL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、72 h后细胞增殖情况;倒置显微镜观察联合作用48 h后细胞集落形态;DAPI染色法、流式细胞仪技术检测48 h时细胞凋亡率;Western blot法检测48 h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),48 h时效果最为明显;荧光显微镜下可见死亡细胞明显减少;Western blot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。

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

Abstract:

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 (P<0.05). CONCLUSION: Fas signaling pathway may be important in the process of SGC-7901 apoptosis induced by combined treatment of As2O3 and TRAIL.

Key words: tumor necrosis factor-related apoptosis-inducing ligand;arsenic trioxide, apoptosis, gastric cancer cells SGC-7901, Fas