期刊文献+
任意字段
题名或关键词
题名
关键词
文摘
作者
第一作者
机构
刊名
分类号
参考文献
作者简介
基金资助
栏目信息

Mcl-1 siRNA对TRAIL诱导胃癌细胞凋亡的影响

Effect of Mcl-1 siRNA on TRAIL-induced apoptosis in gastric cancer cells
下载PDF
导出
摘要 目的探讨转染髓样细胞白血病(Mcl-1)siRNA特异性沉默Mcl-1基因对肿瘤坏死因子相关的凋亡诱导配体(TRAIL)诱导胃癌细胞HGC-27凋亡的影响。方法采用碘化丙啶(PI)染色法流式细胞术分析TRAIL单独作用的细胞凋亡率以及广谱caspase抑制剂z-VAD-fmk对细胞凋亡的抑制作用;Western blot法分析TRAIL处理前后半胱天冬酶-3(caspase-3)和聚腺苷二磷酸核糖聚合酶-1(PARP-1)的活化以及抗凋亡的Bcl-2、Bcl-X_L及Mcl-1的蛋白表达情况;采用Lipofectamine 2000转染Mcl-1 siRNA入HGC-27细胞,Western blot法验证基因沉默效果,流式细胞术分析转染后细胞凋亡率的变化。结果 HGC-27细胞对TRAIL不敏感,48 h的凋亡率仅为(20.65±3.23)%,z-VAD-fmk能几乎完全阻止TRAIL诱导的凋亡;TRAIL作用晚期caspase-3活化、PARP-1裂解,Bcl-2、Bcl-X_L及Mcl-1在TRAIL处理前后没有明显的变化;Mcl-1 siRNA转染后能显著降低细胞中Mcl-1的蛋白表达水平,且细胞转染Mcl-1 siRNA后再加入TRAIL处理,细胞凋亡率明显增加。结论 Mcl-1的高表达可能与HGC-27细胞对TRAIL抵抗有关,Mcl-1 siRNA沉默Mcl-1基因能增加HGC-27细胞对TRAIL的敏感性。 Objective To explore the effect of Mcl-1 small interference RNA （siRNA） on tumor necrosis factor-re- lated apoptosis-inducing ligand （TRAIL）-induced apoptosis in gastric cancer HGC-27 cells. Methods The apop- totic rates of cells treated with TRAIL and pan-caspase inhibitor （z-VAD-fmk） alone or combination were measured by propidium iodide （PI） method using flow cytometry. The activation of caspase-3, cleavage of PARP-1, as well as the protein level of anti-apoptotic Bcl-2 proteins Bcl-2, Bcl-XL and Mcl-1 before and after TRAIL treatment were monitored by Western blot analysis. Transfection of Mcl-1 siRNA was performed using Lipofectamine 2000 reagent. The efficiency of gene silencing was quantified by Western blot and the effect of Mcl-1 siRNA on TRAIL-induced apoptosis was measured using PI method. Results HGC-27 cells were resistant to TRAIL-induced apoptosis, and z-VAD-fmk pretreatment could block apoptosis nearly completely. Activation of caspase-3 and cleavage of PARP-1 occurred in the late stage of apoptosis. The expression levels of Bcl-2, Bcl-XL and Mcl-1 were not altered after ex- posure to TRAIL. Transfection with Mcl-1 siRNA could obviously downregulate the expression level of Mcl-1 in HGC-27 cells and enhanced the sensitivity of cells to TRAIL-induced apoptosis. Conclusion Overexpression of Mcl-1 may account for the resistance of HGC-27 cells to TRAIL. Downregulation of Mcl-1 by siRNA can effectively enhance the sensitivity of HGC-27 ceils to TRAIL-induced apoptosis.
作者 金玮 吴萍
机构地区 安徽医科大学附属巢湖医院耳鼻喉科 安徽医科大学免疫学教研室
出处 《安徽医科大学学报》 CAS 北大核心 2016年第1期47-51,共5页 Acta Universitatis Medicinalis Anhui
基金 安徽高校省级自然科学研究项目(编号:KJ2014A113)
关键词 胃癌 细胞凋亡 小干扰RNA MCL-1 gastric cancer apoptosis siRNA Mcl-1
分类号 R735.2 [医药卫生—肿瘤]
  • 相关文献

参考文献15

  • 1Thomas L W, Lam C, Edwards S W. Mel-1 ; the molecular regu- lation of proteinfunction[J]. FEBS Lett, 2010, 584( 14): 2981 -9.
  • 2Choi E S, 3ung 3 Y, Lee J S, et al. Myeloid cell leukeuda-I is a key molecular target for mithramycin A-induced apoptosis in andro- gen-independent prostate cancer cells and a tumor xenograft animal model[ J ]. Cancer Lett, 2013, 328 ( 1 ) : 65 - 72.
  • 3Skoda C, Erovic B M, Wachek V, et al. Down-regulation of Mcl- 1 with antisense technology alters the effect of various cytotoxic a- gents used in treatment of squamous cell carcinoma of the head and neck[J]. OncolRep, 2008, 19(6): 1499 -503.
  • 4Taniquchi H, Horinaka M, Yoshida T, et al. Targeting the glyox- alase pathway enhances TRAIL efficacy in cancer cells by down- regulating the expression of antiapoptotic molecules [ J ]. Mol Cancer Ther, 2012, 11 (10) : 2294 -300.
  • 5Behnar J, Fesik S W. Small molecule Mcl-I inhibitors for the treatment of cancer [ J]. Pharmaeol Ther, 2015,145:76 - 84.
  • 6Quinn B A, Dash R, Azab B, et al. Targeting Mel-I for the thera- py of cancer [Jl. Expert Opin Investig Drugs, 2011 , 20(10) : 1397 - 411.
  • 7Beroukhim R, Mermel C H, Porter D, et al. The landscape of so- matic copy-number alteration across human cancers [ J] Nature, 2010, 463(7283) :899 -905.
  • 8Wei G, Margolin A A, Haery L, et al. Chemical genonfies identi- fies small-molecule MCL1 repressors and BCL-xL as a predictor of MCL1 dependency [J]. Cancer Cell, 2012, 21(4):547-62.
  • 9吴亚欧,张林杰,张旭东,余宏伟.高表达Mcl-1对衣霉素诱导胃腺癌细胞凋亡的影响[J].安徽医科大学学报,2010,45(1):20-23. 被引量：5
  • 10Wertz l E, Kusam S, Lain C, et al. Sensitivity to antitubulin che- motherapeutics is regulated by MCL1 and FBW7 [ J ]. Nature, 2011,471(7336) : 110 -4.

二级参考文献12

  • 1Rao R V,Ellerby H M, Bredesen D E. Coupling endoplasmic reticulum stress to the cell death program[ J]. Cell Death Differ,2004, 11(4) :372 -80.
  • 2Jiang C C,Lucas K, Avery-Kiejda K A, et al. Up-regulation of Mcl-1 Is Critical for Survival of Human Melanoma Cells upon Endoplasmic Retieulum Stress[ J ]. Cancer Res, 2008,68 (16) :6708 -17.
  • 3Fritsch R M,Schneider G,Saur D, et al. Translational repression of MCL-1 couples stress-induced eIF2 alpha phosphorylation to mitochondrial apoptosis initiation [ J ]. J Biol Chem, 2007,282 ( 31 ) : 22551 - 62.
  • 4Verkhratsky A, Toescu E C. Endoplasmic reticulum Ca^2+ homeostasis and neuronal death [ J ]. J Cell Mol Med,2003,7 (4) :351 - 61.
  • 5Sehroder M, Kaufman R J. ER stress and the unfolded protein response [ J ]. Mutat Res,2005, 569 ( 1 - 2 ) :29 - 31.
  • 6Morishima N,Nakanishi K, Takenouchi H, et al. An endoplasmic reticulum stress-specific caspase cascade in apoptosis. Cytochrome c-independent activation of caspase-9 by caspase-12 [ J]. J Biol Chem ,2002,277 (37) :34287 - 94.
  • 7Koshikawa N, Maejima C, Miyazaki K, et al. Hypoxia selects for high-metastatic Lewis lung carcinoma ceils overexpressing Mcl-1 and exhibiting reduced apoptotic potential in solid tumors[ J]. Oncogene ,2006,25 ( 6 ) :917 - 28.
  • 8Krajewski S, Bodrug S, Krajewska M,et al. Immunohistochemical analysis of Mcl-1 protein in human tissues. Differential regulation of Mcl-1 and Bcl-2 protein production suggests a unique role for Mcl-1 in control of programmed cell death in vivo [ J ]. Am J Pathol, 1995,146 (6) : 1309 - 19.
  • 9Osford S M, Dallman C L, Johnson P W, et al. Current strategies to target the anti-apoptotic Bcl-2 protein in cancer cells[ J]. Curr Med Chem,2004,11(8) :1031 -9.
  • 10Saxena A,Viswanathan S, Moshynska O, et al. Mcl-1 and Bcl-2/ Bax ratio gre associated with treatment response but not with Rai stage in B-cell chronic lymphocytic leukemia[ J]. Am J Hematol, 2004,75 ( 1 ) :22 - 33.

共引文献4

安徽医科大学学报
2016年 第1期

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部