三氧化二砷对人脑胶质瘤干祖细胞的抑制作用及其机制研究
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摘要
目的:研究三氧化二砷(arsenic trioxide,As2O3)对人脑胶质瘤干祖细胞的抑制作用并探讨其机制。
方法:MTT法检测不同浓度As2O3 (3, 6, 12μmo/L)对人脑胶质瘤干祖细胞增殖的影响; Hoechst33258荧光染色、透射电镜观察细胞形态变化;流式细胞仪(PI单染)分析细胞周期变化,流式细胞仪(Annein V/PI双染)检测细胞凋亡率;Western blot检测As2O3与人脑胶质瘤干祖细胞凋亡相关的MLK3/p-MLK3、c-jun/p-c-jun、Fas/FasL蛋白表达变化。
结果:
(1). MTT检测结果显示:3μmo/L As2O3作用人脑胶质瘤干祖细胞24 h,48 h,72 h后的抑制率分别为8.03%,9.96%和21.88%;6μmo/L As2O3作用人脑胶质瘤干祖细胞24 h,48 h,72 h后的抑制率分别为9.43 %,35.80 %和50.40 %;12μmo/L As2O3作用后的抑制率分别为21.88 %,45.47 %和63.10 %。24 h、48 h、72 h的IC50分别为:65.53μmol/L、10.65μmol/L和7.21μmol/L,提示As2O3对人脑胶质瘤干祖细胞的抑制作用呈浓度、时间依赖趋势。
(2). Hoechst33258荧光染色结果显示:对照组细胞核大,Hoechst着色的细胞呈圆形,核内的荧光弱、亮度均一,呈淡兰色。As2O3给药组细胞染色质凝聚、细胞核边集、染色加深等细胞凋亡的形态学改变。
透射电镜观察结果显示:对照组细胞膜表面微绒毛丰富,胞浆量少,细胞核大,核仁明显,核染色质电子密度低,染色质均一;6μmol/L As2O3给药48 h后,细胞膜不完整,细胞膜表面微绒毛和伪足减少,胞质内较多空泡,染色质浓染,细胞核边集,可见凋亡小体。
(3).流式细胞仪凋亡率检测结果显示:0、6、12μmo/L As2O3作用人脑胶质瘤干祖细胞48 h后,凋亡率分别为3.1 %、12.07 %、29.53 %;作用72 h后,凋亡率分别为3.9 %、42.74 %、66.48 %,与对照组相比差异显著(P<0.05)。细胞周期检测结果:对照组G1期细胞占52.153 %,6μmo/L As2O3作用24 h,48 h,72 h后,G1细胞所占比例分别为64.184 %,74.790 %,88.467 %。,提示As2O3可使细胞周期阻滞在G1期。
(4). Western blot检测结果:6μmo/L As2O3处理人脑胶质瘤干祖细胞12 h, 24 h和48 h之后,JNK/Fas信号通路相关蛋白MLK3/p-MLK3、c-jun/p-c-jun、Fas/FasL表达上调,3、6μmo/L As2O3处理48h之后,MLK3/p-MLK3、c-jun/p-c-jun蛋白表达也上调。
结论:As2O3对人脑胶质瘤干祖细胞具有显著的抑制作用,该作用主要与诱导细胞凋亡并使细胞周期阻滞在G1期有关,激活JNK/Fas信号通路可能是其分子机制之一。
Objective: To study the inhitory effect and its mechanism of arsenic trioxide on human glioma stem cells.
Methods: Cell proliferation of As2O3 (3, 6, 12μmo/L) on human glioma stem cells was determined by MTT assay. The morphological changes were observed by Hoechst33258 fluorochrome staining and electron microscope; Cell cycle and apoptosis rate were analysed by flow cytometry. The levels of JNK pathway-mediated MLK3/p-MLK3, c-jun/p-c-jun and Fas/FasL involved in the effect of As2O3 on human glioma stem cells were studied by western blot.
Results:
(1). MTT assay showed that As2O3 had a significant anti-proliferation effect on human glioma stem cells in a dose-dependant and time-dependant manner. After treated with 6μmo/L As2O3 for 24 h, 48 h, 72 h the inhibition rates were 9.43 %,35.80 % and 50.40 %, respectively; After treated with 12μmo/L As2O3 for 24 h, 48 h, 72 h the inhibition rates were 21.88 %,45.47 % and 63.10 %, respectively.
(2). Apoptosis morphological changes about chromatic agglutination and nuclear condensation were detected by Hoechst 33258 fluorochrome staining and electron microscope in As2O3 treated human glioma stem cells.
(3). Apoptosis rates of human gioma stem cells treated with 0, 6, 12μmo/L As2O3 for 48 hours were 3.1 %, 12.07 %, 29.53 %, respectively; After trated with As2O3 for 72 h , the apoptosis rates were 3.9 %, 42.74 % and 66.48 % respectively (P<0.05).
Cell cycle arrest was examined by Flow Cytometry (FCM). After treated with 6μmo/L As2O3 for 24h, 48h, 72h, the G1 phase rates were 64.184 %,74.790 %,88.467 %, respectively. It was 52.153 % in control group.
(4). After treated with 6μmo/L As2O3 for 12h, 24h, 48h the JNK/Fas mediated proteins MLK3/p-MLK3, c-jun/p-c-jun and Fas/FasL were upregulated by western blot. When the cells were treated with 3、6μmo/L As2O3, the MLK3/p-MLK3, c-jun/p-c-jun proteins were also upregulated.
Conclusion: The results suggested that As2O3 could inhibit the proliferation viability of human glioma stem cells (CD133+). It induced cell apoptosis and blocked the cell cycle at G1 phase. The molecule mechanisms may be associated with the activation of JNK/Fas pathway.
方法:MTT法检测不同浓度As2O3 (3, 6, 12μmo/L)对人脑胶质瘤干祖细胞增殖的影响; Hoechst33258荧光染色、透射电镜观察细胞形态变化;流式细胞仪(PI单染)分析细胞周期变化,流式细胞仪(Annein V/PI双染)检测细胞凋亡率;Western blot检测As2O3与人脑胶质瘤干祖细胞凋亡相关的MLK3/p-MLK3、c-jun/p-c-jun、Fas/FasL蛋白表达变化。
结果:
(1). MTT检测结果显示:3μmo/L As2O3作用人脑胶质瘤干祖细胞24 h,48 h,72 h后的抑制率分别为8.03%,9.96%和21.88%;6μmo/L As2O3作用人脑胶质瘤干祖细胞24 h,48 h,72 h后的抑制率分别为9.43 %,35.80 %和50.40 %;12μmo/L As2O3作用后的抑制率分别为21.88 %,45.47 %和63.10 %。24 h、48 h、72 h的IC50分别为:65.53μmol/L、10.65μmol/L和7.21μmol/L,提示As2O3对人脑胶质瘤干祖细胞的抑制作用呈浓度、时间依赖趋势。
(2). Hoechst33258荧光染色结果显示:对照组细胞核大,Hoechst着色的细胞呈圆形,核内的荧光弱、亮度均一,呈淡兰色。As2O3给药组细胞染色质凝聚、细胞核边集、染色加深等细胞凋亡的形态学改变。
透射电镜观察结果显示:对照组细胞膜表面微绒毛丰富,胞浆量少,细胞核大,核仁明显,核染色质电子密度低,染色质均一;6μmol/L As2O3给药48 h后,细胞膜不完整,细胞膜表面微绒毛和伪足减少,胞质内较多空泡,染色质浓染,细胞核边集,可见凋亡小体。
(3).流式细胞仪凋亡率检测结果显示:0、6、12μmo/L As2O3作用人脑胶质瘤干祖细胞48 h后,凋亡率分别为3.1 %、12.07 %、29.53 %;作用72 h后,凋亡率分别为3.9 %、42.74 %、66.48 %,与对照组相比差异显著(P<0.05)。细胞周期检测结果:对照组G1期细胞占52.153 %,6μmo/L As2O3作用24 h,48 h,72 h后,G1细胞所占比例分别为64.184 %,74.790 %,88.467 %。,提示As2O3可使细胞周期阻滞在G1期。
(4). Western blot检测结果:6μmo/L As2O3处理人脑胶质瘤干祖细胞12 h, 24 h和48 h之后,JNK/Fas信号通路相关蛋白MLK3/p-MLK3、c-jun/p-c-jun、Fas/FasL表达上调,3、6μmo/L As2O3处理48h之后,MLK3/p-MLK3、c-jun/p-c-jun蛋白表达也上调。
结论:As2O3对人脑胶质瘤干祖细胞具有显著的抑制作用,该作用主要与诱导细胞凋亡并使细胞周期阻滞在G1期有关,激活JNK/Fas信号通路可能是其分子机制之一。
Objective: To study the inhitory effect and its mechanism of arsenic trioxide on human glioma stem cells.
Methods: Cell proliferation of As2O3 (3, 6, 12μmo/L) on human glioma stem cells was determined by MTT assay. The morphological changes were observed by Hoechst33258 fluorochrome staining and electron microscope; Cell cycle and apoptosis rate were analysed by flow cytometry. The levels of JNK pathway-mediated MLK3/p-MLK3, c-jun/p-c-jun and Fas/FasL involved in the effect of As2O3 on human glioma stem cells were studied by western blot.
Results:
(1). MTT assay showed that As2O3 had a significant anti-proliferation effect on human glioma stem cells in a dose-dependant and time-dependant manner. After treated with 6μmo/L As2O3 for 24 h, 48 h, 72 h the inhibition rates were 9.43 %,35.80 % and 50.40 %, respectively; After treated with 12μmo/L As2O3 for 24 h, 48 h, 72 h the inhibition rates were 21.88 %,45.47 % and 63.10 %, respectively.
(2). Apoptosis morphological changes about chromatic agglutination and nuclear condensation were detected by Hoechst 33258 fluorochrome staining and electron microscope in As2O3 treated human glioma stem cells.
(3). Apoptosis rates of human gioma stem cells treated with 0, 6, 12μmo/L As2O3 for 48 hours were 3.1 %, 12.07 %, 29.53 %, respectively; After trated with As2O3 for 72 h , the apoptosis rates were 3.9 %, 42.74 % and 66.48 % respectively (P<0.05).
Cell cycle arrest was examined by Flow Cytometry (FCM). After treated with 6μmo/L As2O3 for 24h, 48h, 72h, the G1 phase rates were 64.184 %,74.790 %,88.467 %, respectively. It was 52.153 % in control group.
(4). After treated with 6μmo/L As2O3 for 12h, 24h, 48h the JNK/Fas mediated proteins MLK3/p-MLK3, c-jun/p-c-jun and Fas/FasL were upregulated by western blot. When the cells were treated with 3、6μmo/L As2O3, the MLK3/p-MLK3, c-jun/p-c-jun proteins were also upregulated.
Conclusion: The results suggested that As2O3 could inhibit the proliferation viability of human glioma stem cells (CD133+). It induced cell apoptosis and blocked the cell cycle at G1 phase. The molecule mechanisms may be associated with the activation of JNK/Fas pathway.
引文
[1]. Claus EB, Black PM. Survival rates and patterns of care for patients diagnosed with supratentorial low-grade gliomas:Data from the SEER program, 1973-2001[J].Cancer, 2006,106(12): 1358-1363.
[2]. Farquhar D, Pan BF, Sakurai M, Ghost A, Mullen CA, Nelson JA. Suicide gene therapy using E.coliβ-galactosidase [J]. Cancer Chemother Pharmacol, 2002; 50(1): 65-70.
[3].姜华,李支援,张东君.大脑胶质瘤病[J].国际内科学杂志,2009,36(6):343-345.
[4].马国峡.脑胶质瘤的手术治疗探讨[J].临床探讨, 2009,47(14).
[5].付尚志.脑胶质瘤放射治疗实验及临床研究进展[J].临床军医杂志,2008,36(2).
[6].高岩升,王凯娟.胶质瘤治疗策略[J].医学信息(内·外科版),2009,22(4).
[7]. Hess KR, Broglio KR, Bondy ML. Adult glioma incidence in the United State, 1977-2000 [J]. Cancer, 2004; 101(10): 2293-2299.
[8].赵继宗.颅脑肿瘤外科学[M].北京人民卫生出版社,2004:208-234.
[9]. Faith GD, Sally F, Tames G, et al. Survival rates in patients withprimary malignant brain tumors stratified by patient age and tu-mor histological type:an analysis based on Surveillance, epidemicology, and end results (SEER) data [J]. J Neurosurg, 1998,88(1):1973-1991.
[10]. Bonnet D, Dick JE. Human acute myeloid leukemia is organized as a hieraccchy that originates from a primitive hematopoietic cell [J]. Nat Med., 1997, 3(7):730-737.
[11]. Cox CV, Evely RS, Oakhill A, et al. Characterization of acute lymphoblastic leukemia progenitor cell [J]. Blood,2004,104(9):2919.
[12]. Aguado T, Carracedo A, Julien B, et al. Cannabinoids induce glioma stem-like cell differentiation and inhibit glimagenesis[J]. J Biol Chem., 2007, 282 (9):6854-6862.
[13].于春泳,杨辉.神经干细胞与脑胶质瘤干细胞[J].中国组织工程研究与临床康复,2008.
[14]. Zhu Q, Zhang JW, Zhu HQ, et al. Synergic effects of arsenic trioxide and cAMP during acute promyelocytic leukemia cell maturation subtends a novel signaling cross-talk [J/OL]. Blood, 2002,99(3):1014-1022.
[15]. Cai X, Shen YL, Zhu Q, et al. Arsenic trioxide-induced apoptosis and differentiation are associated respectively with mitochondrial transmembrane potential collapse and retinoic acid signaling pathways in acute promyelocytic leukemia [J]. Leukemia, 2000,14(2):262-270.
[16]. Singh SK, Hawkins C, Clarke ID, et al. Identification of human brain tumour initiating cells[J]. Nature,2004, 432(7015): 396-401.
[17]. Kondo T, Setoguchi T, Taga T, et al. Persistence of a small subpopulation of cancer stem-like cells in the C6 glioma cell line[J]. Proc Natl Acad Sci, 2004, 101(12):781-786.
[18].赵耀东,黄强,张天一等.胶质瘤干-祖细胞超微结构分析[J].中华肿瘤杂志,2008,30(9):663-667.
[19]. Yaodong Zhao, Qiang Huang, Tianyi Zhang, et al. Ultrastructural Studies of Glioma Stem Cells/Progenitor Cells [J]. Ultrastructural Pathology,2008,32: 241–245.
[20]. Bao SD ,Wu QL ,Mclendon R , et al. Glioma stem cells promote radioresistance by preferential activation of the DNA damage response [ J ] . Nature,2006,444 (7120 ):756.
[21]. Ragione FD, Borriello A, Giordani L, et al. Cell division cycle alterations in human malignancies[J]. Cancer,1997,10(1):151-156.
[22]. Jia p, Chen G, Huang X, et al. Arsenic trioxide induces multiple myeloma cell apoptosis via disruption of mitochondrial transmembrane potentials and activation of capase-3 [J]. Chin Med J (Engl), 2001,114(1):19-24.
[23].邢茂,张思娟,叶鑫等.三氧化二砷诱导肿瘤细胞凋亡途径的研究[J].中国药理学通报, 2002,18(1):87-90.
[24].王丽,陈治文,夏俊等.三氧化二砷对黑色素瘤B16细胞端粒酶活性的影响[J].蚌埠医学院学报, 2004,29(3):202-204.
[25]. Salven P.Angiogenesis in lymphoproliferative disorders [J]. Acta Haematol, 2001,106(4):184-189.
[26]. Roboz GJ, Dias S, Lam G, et al. Arsenic trioxide induces dose-and time-dependent apoptosis of endothelium and may exert an antileukemic effect via inhibition of angiogenesis[J]. Blood, 2000, 96(4):1525-1530.
[27]. LOOK A T. Aresnic and apoptosis in the treatment of acute promyelocytic leukemia[J]. J Natl Cancer Inst, 1998,90(2):86.
[28]. Seol JG, Park WH, Kim ES, et al. Effect of arsenic trioxide on cell cycle arrest in head and neck cancer cell line PCI-1[J]. Biochem Biophys Res Commun, 1999,265(2):400-404.
[29]. Bazarbachi A, El Sabban ME, Nasr R, et al. Arsenic trioxide and interferon-alpha synergize to induce cell cycle arrest and apoptosis inhuman T-cell lymphotropic virus type I-transformed cells[J]. Blood,1999,93(1): 278 -283.
[30].张鹏,王树叶,胡龙虎等.三氧化二砷注射液治疗72例急性早幼粒细胞白血病[J].中华血液学杂志, 1996,17(2):58-60.
[31].詹梦熊,林秋泉,冯华,朱刚.胶质瘤干细胞研究进展[J].中国临床神经外科杂志,2007,14(5):315-318.
[32].于春泳,杨辉.神经干细胞与脑胶质瘤干细胞[J].山东医药,2008,48(37): 112-113.
[33]. Shen ZY, Shen WY, ChenMH, et al. Mitochondria,calcium and nitric oxide in the apoptotic pathway of esophageal carcinoma cells induced by As2O3[J]. Int,88.
[34].黄强.脑肿瘤干细胞分化抑制分子机制研究[J].Chin J Neurosurg Dis Res,2007,6(4).
[35].Antonyak MA, Kenyon LC, Godwin AK, et al. Elevated JNK activation contributes to the pathogenesis of human brain tumors[J]. Oncogene.,2002, 21(33):5038-46.
[36]. Ouafik L, Berenguer-Daize C, Berthois Y. Adrenomedullin promotes cell cycle transit and up-regulates cyclin D1 protein level in human glioblastoma cells throughthe activation of c-Jun/JNK/AP-1 signal transduction pathway[J]. Cell Signal.,2009,21(4):597-608.
[37]. WILSON H. MILLER, JR. Molecular Targets of Arsenic Trioxide in Malignant Cells[J]. Oncologist., 2002,7(1):14-19.
[38]. Kanzawa T, Iwado E, Aoki H., et al. Ionizing radiation induces apoptosis and inhibits neuronal differentiation in rat neural stem cells via the c-Jun NH2-terminal kinase (JNK) pathway[J]. Oncogene. 2006,22;25(26):3638-3648.
[39].李玉林,唐建武等,病理学,第六版,人民卫生出版社.
[40].王静,王尉平,顾振纶,梁中琴,周文轩,郭次仪.熊果酸诱导肺腺癌SPC-A-1细胞凋亡及其机制[J].苏州大学学报:医学版,2008,2:207-210.
[41]. Qiang Huang, Quan-Bin Zhang, Jun Dong, and et al. Glioma stem cells are more aggressive in recurrent tumors with malignant progression than in the primary tumor, and both can be maintained long-term in vitro[J]. BMC Cancer 2008, 8:304.
[42].肖玉强.三氧化二砷诱导C6胶质瘤细胞凋亡实验研究[J].中华神经外科疾病研究杂志,2007,4.
[43].魏玲,王兴武.三氧化二砷抑制人神经胶质瘤细胞生长及其机制[J].中国病理生理杂志,2006,4.
[44]. Zhao S, Tsuchida T, Kawakami K, et al. Effect of As2O3 on cell cycle progression and cyclins D1 and B1 expression in two glioblastoma cell lines differing in p53 status[J]. Int J Oncol., 2002,21(1):49-55.
[45]. Wei Y, Liu D, Ge Y, et al. Down-regulation of beta1,4GalT V at protein level contributes to arsenic trioxide-induced glioma cell apoptosis[J]. Cancer Lett., 2008,18;267(1):96-105.
[46]. Zhao S, Zhang J, Zhang X, et al. Arsenic trioxide induces different gene expression profiles of genes related to growth and apoptosis in glioma cells dependent on the p53 status[J]. 2008, 35(3):421-9.
[47]. Cheng Y, Chang LW, Tsou L. Mitogen-activated protein kinases mediatedarsenic-induced down-regulation of survivin in human lung adenocarcinoma cells. Arch Toxicol. 2006; 80 (6):310-8.
[48]. Han YH, Kim SZ, Kim SH, et al. Induction of apoptosis in arsenic trioxide-treated lung cancer A549 cells by buthiomine sulfoximine. Mol Cells. 2008; 26 (2): 158-64.
[49]. Houtgraaf JH, Versmissen J, van der Giessen WJ. A concise review of DNA damage checkpoints and repair in mammalian cells. Cardiovasc Revasc Med. 2006,7 (3): 165-72.
[50]. Schwartz GK, Shah MA. Targeting the cell cycle: a new approach to cancer therapy. J Clin Oncol. 2005, 23(36):9408-9421.
[51]. Nakamura Y, Miyoshi N. Cell death induction by isothiocyanate and their underlying molecular mechanisms. Biofactors. 2006,26 (2):123-134.
[52]. Zhao S, Tsuchida T, Kawakami K, et al. Effect of As2O3 on cell cycle progression and cyclins D1 and B1 expression in two glioblastoma cell lines differing in p53 status. Int J Oncol.,2002,21(1): 49-55.
[53]. Shao QS, Ye ZY, Ling ZQ, et al. Cell cycle arrest and apoptotic cell death in cultured human gastric carcinoma cells mediated by arsenic trioxide. World J Gastroenterol. 2005,11(22):3451-3456.
[54]. Houtgraaf JH, Versmissen J, van der Giessen WJ. A concise review of DNA damage checkpoints and repair in mammalian cells. Cardiovasc Revasc Med. 2006,7 (3): 165-172.
[55].张秀娟,李秋实,季宇彬.细胞死亡方式研究进展[J].哈尔滨医药, 2007,06.
[56]. Williams GT. Programmed cell deah: apoptosis and oncogenesis [J]. Cell,1991, 65(7):1097-1098.
[57].叶冬青,高维娟. c-jun氨基末端激酶信号通路与细胞凋亡[J].中国老年学杂志,2009,29:894-896.
[58]. Minden A,LinA,Claret Fx,et a1.Selective activation of the jnk signaling cascade and c-jun transcriptional activity by the small GTPases Rac and cdc-42Hs [J].cell,1995,81(7):1147.
[59]. Weitzman JB.JNK [J].Curr Biol,2000,10(8):R290.
[60]. Fuchs SY,Adler V,Buschmann T,et a1.JNK targets p53 ubiquitination and degradation in nonstressed cells [J].Gene Dev,1998,12(1):2658-2663,
[61]. Kennedy NJ, Davis RJ: Role of JNK in tumor development [J]. Cell Cycle, 2003,2:199-201.
[62]. Uhlirova M, Jasper H, Bohmann D: Non-cell-autonomous induction of tissue overgrowth by JNK/Ras cooperation in a Drosophila tumor model [J]. Proc Natl Acad Sci USA,2005,102:13123-13128.
[63]. Nateri AS, Spencer-Dene B, Behrens A: Interaction of phosphorylated c-Jun with TCF4 regulates intestinal cancer development [J]. Nature,2005,437:281 -285.
[64].李国荣,朱婉儿. Fas/FasL系统与中枢神经系统疾病[J].中国实用神经系统疾病杂志,2007,10(8): 133-137.
[65].熊小平,杨毅细胞凋亡及Fas/FasL、Bcl-2与重症肌无力[J].中国临床保健杂志,2008,11(5):542-544.
[66]. A.E.Konstantinidou, P.Korkolopoulou, E. Patsouris. Apoptotic markers for primary brain tumor prognosis [J]. Journal of Neuro-Oncologu.,2005,72: 151-156.
[67]. Bertrand J, Begaud-Grimaud G, Bessette B, et al. Cancer stem cells from human glioma cell line are resistant to Fas-induced apoptosis[J]. Int J Oncol., 2009,34(3):717-27.
[68]. Mazar J, Thomas M, Bezrukov L, et al. Cytotoxicity mediated by the Fas ligand (FasL)-activated apoptotic pathway in stem cells [J]. J Biol Chem.,2009,284 (33):22022-22028.
[1]秦玉新,蒙凌华,丁健. RNA干扰技术的研究进展[J].中国药理学通报,2007,23(4):421-4.
[1] QIN Y X,MENG L H,Ding J. Research progresses of RNA interference[J]. Chin Pharmacol Bull, 2007,21(8):901-4.
[2] Rosalind C L , Rhonda L F, Victor A. The C. elegans heterochronic gene lin-4 encodessmall RNAs with antisense complementarity to lin-14[J]. Cell,1993, 12(5):843-854.
[3] Reinhart B J, Slack F J, Basson M, et al. The 21-nucleotide let-7 RNA regulates developmental timing in Caenorhabditis elegans[J]. Nature, 2000,403 :901–906.
[4] Griffiths J S, Grocock R J, Van D S, et al. MiRBase: microRNA sequences, targets and gene nomenclature[J]. Nucleic Acids Res,2006 ,1(34):D140-4.
[5] Cimmino A, Calin GA, Fabbri M, et al. MiR-15 and miR-16 induce apoptosis by targeting BCL2[J]. Proc Natl Acad Sci U S A. 2005,102(39):13944-9.
[6] Xia L, Zhang D, Du R, et al. miR-15b and miR-16 modulate multidrug resistance by targeting BCL2 in human gastric cancer cells[J]. Int J Cancer. 2008,123(2):372-9.
[7] Linsley S., Schelter J, Burchard J,et al. Transcripts Targeted by the MicroRNA-16 Family Cooperatively Regulate Cell Cycle Progression[J]. Molecular and biology. 2007,3(27): 2240–2252.
[8] He X, He L, Hannon G J. The guardian's little helper: microRNAs in the p53 tumor suppressor network. [J]. Cancer Res, 2007,67(23):11099-101.
[9] Gironella M, Seux M, Xie M J, et al. Tumor protein53-induced nuclear protein 1 expression is repressed by miR-155, and its restoration inhibits pancreatic tumor development[J]. Proc Natl Acad Sci U S A, 2007 ,104(41):16170-5.
[10] Xi Y, Formentini A, Chien M,et al. Prognostic Values of microRNAs in Colorectal Cancer[J]. Biomark Insights. 2006,2: 113-121
[11] Ovcharenko D, Kelnar K, Johnson C, et al. Genome-scale microRNA and small interfering RNA screens identify small RNA modulators of TRAIL-induced apoptosis pathway[J]. Cancer Res, 2007,67(22):10782-8.
[12] Corsten M F, Miranda R, Kasmieh R, et al. MicroRNA-21 knockdown disrupts glioma growth in vivo and displays synergistic cytotoxicity with neural precursor cell delivered S-TRAIL in human gliomas[J]. Cancer Res, 2007,67 (19): 8994 -9000.
[13]王贞丽,李金莲,王春波.肿瘤坏死因子相关凋亡诱导配体及扇贝多肽在UVA诱导HaCaT细胞凋亡中的作用[J].中国药理学通报,2007,23(10):1375～9.
[13] Wang Zh L, Li J L, Wang Ch B. The role of tumor necrosis factor related apoptosis inducing ligand and Polypeptide from Chlamys farreri in UVA-induced apoptosis in HaCaT cells[J]. Chin Pharmacol Bull, 2007 ,23(10):1375～9.
[14] Sampson V B, Rong N H, Han J, et al. MicroRNA let-7a down-regulates MYC and reverts MYC-induced growth in Burkitt lymphoma cells[J]. Cancer Res, 2007,67(20):9762-70.
[15] Akao Y, Nakagawa Y, Naoe T. Let-7 microRNA functions as a potential growth suppressor in human colon cancer cells[J]. Biol Pharm Bull,2006,29(5):903-6.
[16] O'Donnell K A, Wentzel E A, Zeller K I, et al. C-Myc-regulated microRNAs modulate E2F1 expression[J]. Nature, 2005,435 (7043):839-43.
[17] He L, Thomson J., Hemann M,et al. A microRNA polycistron as a potential human Oncogene[J]. Nature, 2005,435:828-833.
[18] Sylvestre Y, De Guire V, Querido E, et al. An E2F/miR-20a autoregulatory feedback loop[J]. J Biol Chem, 2007,282 (4): 2135-43.
[19] Tazawa H, Tsuchiya N, Izumiya M, et al. Tumor-suppressive miR-34a induces senescence-like growth arrest through modulation of the E2F pathway in human colon cancer cells[J]. Proc Natl Acad Sci USA,2007,104(39):15472-7.
[20] Yu F Y, Yao H, Zhu P C, et al. Let-7 Regulates self Renewal and Tumorigenicity of Breast Cancer Cells[J]. Cell,2007,12 (6): 1109 -1123.
[21] Hilah G, Gopal P, Andrew A., et al. MIR-451 and Imatinib mesylate inhibit tumor growth of Glioblastoma stem cells[J]. Biochemical and Biophysical Research Communications,2008,8 (31):86–9.
[22] Silber J,Lim D,Petritsch C,et al. MiR-124 and miR-137 inhibit proliferation of glioblastoma multiforme cells and induce differentiation of brain tumor stem cells[J]. BMC Medicine, 2008,6:14.
[23] Neumuller R, Betschinger J, Fischer A,et al. Mei-P26 regulates microRNAs and cell growth in the Drosophila ovarian stem cell lineage[J]. Nature,2008, 454(7201):241-54.
[24]黎丹戎,张汉英,张玮等.汉黄芩素对人卵巢癌细胞株SKOV3裸鼠移植瘤生长及端粒酶活性的抑制作用[J].中国药理学通报, 2007,23(4):119-123.
[24] LI D R,ZHANG H Y,ZHANG W, et al. The inhibitory role of wogonin on tumor growth and telomerase activity of human ovarian cancer SKOV3 cell line xenograf in nude mice[J]. Chinese Pharmacological Bulletin,2007, 23(4): 119-123.
[25]林红英,殷润婷,奚涛等.在肿瘤研究中转基因小鼠模型的研究进展[J].中国药理学通报,2007,23(1):4-8.
[25] LIN H Y, YIN R T,XI T ,et al. Transgenic mouse models of inhibition of tumor angiogenesis[J]. Chinese Pharmacological Bulletin, 2007,23(1):4-8.
[26] Liang Z X , Wu H, Santosh Reddy , et al. Blockade of invasion and metastasis of breast cancer cells via targeting CXCR4 with an artificial microRNA[J]. Biochemical and Biophysical Research Communications, 2007,363:542–546.
[2]. Farquhar D, Pan BF, Sakurai M, Ghost A, Mullen CA, Nelson JA. Suicide gene therapy using E.coliβ-galactosidase [J]. Cancer Chemother Pharmacol, 2002; 50(1): 65-70.
[3].姜华,李支援,张东君.大脑胶质瘤病[J].国际内科学杂志,2009,36(6):343-345.
[4].马国峡.脑胶质瘤的手术治疗探讨[J].临床探讨, 2009,47(14).
[5].付尚志.脑胶质瘤放射治疗实验及临床研究进展[J].临床军医杂志,2008,36(2).
[6].高岩升,王凯娟.胶质瘤治疗策略[J].医学信息(内·外科版),2009,22(4).
[7]. Hess KR, Broglio KR, Bondy ML. Adult glioma incidence in the United State, 1977-2000 [J]. Cancer, 2004; 101(10): 2293-2299.
[8].赵继宗.颅脑肿瘤外科学[M].北京人民卫生出版社,2004:208-234.
[9]. Faith GD, Sally F, Tames G, et al. Survival rates in patients withprimary malignant brain tumors stratified by patient age and tu-mor histological type:an analysis based on Surveillance, epidemicology, and end results (SEER) data [J]. J Neurosurg, 1998,88(1):1973-1991.
[10]. Bonnet D, Dick JE. Human acute myeloid leukemia is organized as a hieraccchy that originates from a primitive hematopoietic cell [J]. Nat Med., 1997, 3(7):730-737.
[11]. Cox CV, Evely RS, Oakhill A, et al. Characterization of acute lymphoblastic leukemia progenitor cell [J]. Blood,2004,104(9):2919.
[12]. Aguado T, Carracedo A, Julien B, et al. Cannabinoids induce glioma stem-like cell differentiation and inhibit glimagenesis[J]. J Biol Chem., 2007, 282 (9):6854-6862.
[13].于春泳,杨辉.神经干细胞与脑胶质瘤干细胞[J].中国组织工程研究与临床康复,2008.
[14]. Zhu Q, Zhang JW, Zhu HQ, et al. Synergic effects of arsenic trioxide and cAMP during acute promyelocytic leukemia cell maturation subtends a novel signaling cross-talk [J/OL]. Blood, 2002,99(3):1014-1022.
[15]. Cai X, Shen YL, Zhu Q, et al. Arsenic trioxide-induced apoptosis and differentiation are associated respectively with mitochondrial transmembrane potential collapse and retinoic acid signaling pathways in acute promyelocytic leukemia [J]. Leukemia, 2000,14(2):262-270.
[16]. Singh SK, Hawkins C, Clarke ID, et al. Identification of human brain tumour initiating cells[J]. Nature,2004, 432(7015): 396-401.
[17]. Kondo T, Setoguchi T, Taga T, et al. Persistence of a small subpopulation of cancer stem-like cells in the C6 glioma cell line[J]. Proc Natl Acad Sci, 2004, 101(12):781-786.
[18].赵耀东,黄强,张天一等.胶质瘤干-祖细胞超微结构分析[J].中华肿瘤杂志,2008,30(9):663-667.
[19]. Yaodong Zhao, Qiang Huang, Tianyi Zhang, et al. Ultrastructural Studies of Glioma Stem Cells/Progenitor Cells [J]. Ultrastructural Pathology,2008,32: 241–245.
[20]. Bao SD ,Wu QL ,Mclendon R , et al. Glioma stem cells promote radioresistance by preferential activation of the DNA damage response [ J ] . Nature,2006,444 (7120 ):756.
[21]. Ragione FD, Borriello A, Giordani L, et al. Cell division cycle alterations in human malignancies[J]. Cancer,1997,10(1):151-156.
[22]. Jia p, Chen G, Huang X, et al. Arsenic trioxide induces multiple myeloma cell apoptosis via disruption of mitochondrial transmembrane potentials and activation of capase-3 [J]. Chin Med J (Engl), 2001,114(1):19-24.
[23].邢茂,张思娟,叶鑫等.三氧化二砷诱导肿瘤细胞凋亡途径的研究[J].中国药理学通报, 2002,18(1):87-90.
[24].王丽,陈治文,夏俊等.三氧化二砷对黑色素瘤B16细胞端粒酶活性的影响[J].蚌埠医学院学报, 2004,29(3):202-204.
[25]. Salven P.Angiogenesis in lymphoproliferative disorders [J]. Acta Haematol, 2001,106(4):184-189.
[26]. Roboz GJ, Dias S, Lam G, et al. Arsenic trioxide induces dose-and time-dependent apoptosis of endothelium and may exert an antileukemic effect via inhibition of angiogenesis[J]. Blood, 2000, 96(4):1525-1530.
[27]. LOOK A T. Aresnic and apoptosis in the treatment of acute promyelocytic leukemia[J]. J Natl Cancer Inst, 1998,90(2):86.
[28]. Seol JG, Park WH, Kim ES, et al. Effect of arsenic trioxide on cell cycle arrest in head and neck cancer cell line PCI-1[J]. Biochem Biophys Res Commun, 1999,265(2):400-404.
[29]. Bazarbachi A, El Sabban ME, Nasr R, et al. Arsenic trioxide and interferon-alpha synergize to induce cell cycle arrest and apoptosis inhuman T-cell lymphotropic virus type I-transformed cells[J]. Blood,1999,93(1): 278 -283.
[30].张鹏,王树叶,胡龙虎等.三氧化二砷注射液治疗72例急性早幼粒细胞白血病[J].中华血液学杂志, 1996,17(2):58-60.
[31].詹梦熊,林秋泉,冯华,朱刚.胶质瘤干细胞研究进展[J].中国临床神经外科杂志,2007,14(5):315-318.
[32].于春泳,杨辉.神经干细胞与脑胶质瘤干细胞[J].山东医药,2008,48(37): 112-113.
[33]. Shen ZY, Shen WY, ChenMH, et al. Mitochondria,calcium and nitric oxide in the apoptotic pathway of esophageal carcinoma cells induced by As2O3[J]. Int,88.
[34].黄强.脑肿瘤干细胞分化抑制分子机制研究[J].Chin J Neurosurg Dis Res,2007,6(4).
[35].Antonyak MA, Kenyon LC, Godwin AK, et al. Elevated JNK activation contributes to the pathogenesis of human brain tumors[J]. Oncogene.,2002, 21(33):5038-46.
[36]. Ouafik L, Berenguer-Daize C, Berthois Y. Adrenomedullin promotes cell cycle transit and up-regulates cyclin D1 protein level in human glioblastoma cells throughthe activation of c-Jun/JNK/AP-1 signal transduction pathway[J]. Cell Signal.,2009,21(4):597-608.
[37]. WILSON H. MILLER, JR. Molecular Targets of Arsenic Trioxide in Malignant Cells[J]. Oncologist., 2002,7(1):14-19.
[38]. Kanzawa T, Iwado E, Aoki H., et al. Ionizing radiation induces apoptosis and inhibits neuronal differentiation in rat neural stem cells via the c-Jun NH2-terminal kinase (JNK) pathway[J]. Oncogene. 2006,22;25(26):3638-3648.
[39].李玉林,唐建武等,病理学,第六版,人民卫生出版社.
[40].王静,王尉平,顾振纶,梁中琴,周文轩,郭次仪.熊果酸诱导肺腺癌SPC-A-1细胞凋亡及其机制[J].苏州大学学报:医学版,2008,2:207-210.
[41]. Qiang Huang, Quan-Bin Zhang, Jun Dong, and et al. Glioma stem cells are more aggressive in recurrent tumors with malignant progression than in the primary tumor, and both can be maintained long-term in vitro[J]. BMC Cancer 2008, 8:304.
[42].肖玉强.三氧化二砷诱导C6胶质瘤细胞凋亡实验研究[J].中华神经外科疾病研究杂志,2007,4.
[43].魏玲,王兴武.三氧化二砷抑制人神经胶质瘤细胞生长及其机制[J].中国病理生理杂志,2006,4.
[44]. Zhao S, Tsuchida T, Kawakami K, et al. Effect of As2O3 on cell cycle progression and cyclins D1 and B1 expression in two glioblastoma cell lines differing in p53 status[J]. Int J Oncol., 2002,21(1):49-55.
[45]. Wei Y, Liu D, Ge Y, et al. Down-regulation of beta1,4GalT V at protein level contributes to arsenic trioxide-induced glioma cell apoptosis[J]. Cancer Lett., 2008,18;267(1):96-105.
[46]. Zhao S, Zhang J, Zhang X, et al. Arsenic trioxide induces different gene expression profiles of genes related to growth and apoptosis in glioma cells dependent on the p53 status[J]. 2008, 35(3):421-9.
[47]. Cheng Y, Chang LW, Tsou L. Mitogen-activated protein kinases mediatedarsenic-induced down-regulation of survivin in human lung adenocarcinoma cells. Arch Toxicol. 2006; 80 (6):310-8.
[48]. Han YH, Kim SZ, Kim SH, et al. Induction of apoptosis in arsenic trioxide-treated lung cancer A549 cells by buthiomine sulfoximine. Mol Cells. 2008; 26 (2): 158-64.
[49]. Houtgraaf JH, Versmissen J, van der Giessen WJ. A concise review of DNA damage checkpoints and repair in mammalian cells. Cardiovasc Revasc Med. 2006,7 (3): 165-72.
[50]. Schwartz GK, Shah MA. Targeting the cell cycle: a new approach to cancer therapy. J Clin Oncol. 2005, 23(36):9408-9421.
[51]. Nakamura Y, Miyoshi N. Cell death induction by isothiocyanate and their underlying molecular mechanisms. Biofactors. 2006,26 (2):123-134.
[52]. Zhao S, Tsuchida T, Kawakami K, et al. Effect of As2O3 on cell cycle progression and cyclins D1 and B1 expression in two glioblastoma cell lines differing in p53 status. Int J Oncol.,2002,21(1): 49-55.
[53]. Shao QS, Ye ZY, Ling ZQ, et al. Cell cycle arrest and apoptotic cell death in cultured human gastric carcinoma cells mediated by arsenic trioxide. World J Gastroenterol. 2005,11(22):3451-3456.
[54]. Houtgraaf JH, Versmissen J, van der Giessen WJ. A concise review of DNA damage checkpoints and repair in mammalian cells. Cardiovasc Revasc Med. 2006,7 (3): 165-172.
[55].张秀娟,李秋实,季宇彬.细胞死亡方式研究进展[J].哈尔滨医药, 2007,06.
[56]. Williams GT. Programmed cell deah: apoptosis and oncogenesis [J]. Cell,1991, 65(7):1097-1098.
[57].叶冬青,高维娟. c-jun氨基末端激酶信号通路与细胞凋亡[J].中国老年学杂志,2009,29:894-896.
[58]. Minden A,LinA,Claret Fx,et a1.Selective activation of the jnk signaling cascade and c-jun transcriptional activity by the small GTPases Rac and cdc-42Hs [J].cell,1995,81(7):1147.
[59]. Weitzman JB.JNK [J].Curr Biol,2000,10(8):R290.
[60]. Fuchs SY,Adler V,Buschmann T,et a1.JNK targets p53 ubiquitination and degradation in nonstressed cells [J].Gene Dev,1998,12(1):2658-2663,
[61]. Kennedy NJ, Davis RJ: Role of JNK in tumor development [J]. Cell Cycle, 2003,2:199-201.
[62]. Uhlirova M, Jasper H, Bohmann D: Non-cell-autonomous induction of tissue overgrowth by JNK/Ras cooperation in a Drosophila tumor model [J]. Proc Natl Acad Sci USA,2005,102:13123-13128.
[63]. Nateri AS, Spencer-Dene B, Behrens A: Interaction of phosphorylated c-Jun with TCF4 regulates intestinal cancer development [J]. Nature,2005,437:281 -285.
[64].李国荣,朱婉儿. Fas/FasL系统与中枢神经系统疾病[J].中国实用神经系统疾病杂志,2007,10(8): 133-137.
[65].熊小平,杨毅细胞凋亡及Fas/FasL、Bcl-2与重症肌无力[J].中国临床保健杂志,2008,11(5):542-544.
[66]. A.E.Konstantinidou, P.Korkolopoulou, E. Patsouris. Apoptotic markers for primary brain tumor prognosis [J]. Journal of Neuro-Oncologu.,2005,72: 151-156.
[67]. Bertrand J, Begaud-Grimaud G, Bessette B, et al. Cancer stem cells from human glioma cell line are resistant to Fas-induced apoptosis[J]. Int J Oncol., 2009,34(3):717-27.
[68]. Mazar J, Thomas M, Bezrukov L, et al. Cytotoxicity mediated by the Fas ligand (FasL)-activated apoptotic pathway in stem cells [J]. J Biol Chem.,2009,284 (33):22022-22028.
[1]秦玉新,蒙凌华,丁健. RNA干扰技术的研究进展[J].中国药理学通报,2007,23(4):421-4.
[1] QIN Y X,MENG L H,Ding J. Research progresses of RNA interference[J]. Chin Pharmacol Bull, 2007,21(8):901-4.
[2] Rosalind C L , Rhonda L F, Victor A. The C. elegans heterochronic gene lin-4 encodessmall RNAs with antisense complementarity to lin-14[J]. Cell,1993, 12(5):843-854.
[3] Reinhart B J, Slack F J, Basson M, et al. The 21-nucleotide let-7 RNA regulates developmental timing in Caenorhabditis elegans[J]. Nature, 2000,403 :901–906.
[4] Griffiths J S, Grocock R J, Van D S, et al. MiRBase: microRNA sequences, targets and gene nomenclature[J]. Nucleic Acids Res,2006 ,1(34):D140-4.
[5] Cimmino A, Calin GA, Fabbri M, et al. MiR-15 and miR-16 induce apoptosis by targeting BCL2[J]. Proc Natl Acad Sci U S A. 2005,102(39):13944-9.
[6] Xia L, Zhang D, Du R, et al. miR-15b and miR-16 modulate multidrug resistance by targeting BCL2 in human gastric cancer cells[J]. Int J Cancer. 2008,123(2):372-9.
[7] Linsley S., Schelter J, Burchard J,et al. Transcripts Targeted by the MicroRNA-16 Family Cooperatively Regulate Cell Cycle Progression[J]. Molecular and biology. 2007,3(27): 2240–2252.
[8] He X, He L, Hannon G J. The guardian's little helper: microRNAs in the p53 tumor suppressor network. [J]. Cancer Res, 2007,67(23):11099-101.
[9] Gironella M, Seux M, Xie M J, et al. Tumor protein53-induced nuclear protein 1 expression is repressed by miR-155, and its restoration inhibits pancreatic tumor development[J]. Proc Natl Acad Sci U S A, 2007 ,104(41):16170-5.
[10] Xi Y, Formentini A, Chien M,et al. Prognostic Values of microRNAs in Colorectal Cancer[J]. Biomark Insights. 2006,2: 113-121
[11] Ovcharenko D, Kelnar K, Johnson C, et al. Genome-scale microRNA and small interfering RNA screens identify small RNA modulators of TRAIL-induced apoptosis pathway[J]. Cancer Res, 2007,67(22):10782-8.
[12] Corsten M F, Miranda R, Kasmieh R, et al. MicroRNA-21 knockdown disrupts glioma growth in vivo and displays synergistic cytotoxicity with neural precursor cell delivered S-TRAIL in human gliomas[J]. Cancer Res, 2007,67 (19): 8994 -9000.
[13]王贞丽,李金莲,王春波.肿瘤坏死因子相关凋亡诱导配体及扇贝多肽在UVA诱导HaCaT细胞凋亡中的作用[J].中国药理学通报,2007,23(10):1375～9.
[13] Wang Zh L, Li J L, Wang Ch B. The role of tumor necrosis factor related apoptosis inducing ligand and Polypeptide from Chlamys farreri in UVA-induced apoptosis in HaCaT cells[J]. Chin Pharmacol Bull, 2007 ,23(10):1375～9.
[14] Sampson V B, Rong N H, Han J, et al. MicroRNA let-7a down-regulates MYC and reverts MYC-induced growth in Burkitt lymphoma cells[J]. Cancer Res, 2007,67(20):9762-70.
[15] Akao Y, Nakagawa Y, Naoe T. Let-7 microRNA functions as a potential growth suppressor in human colon cancer cells[J]. Biol Pharm Bull,2006,29(5):903-6.
[16] O'Donnell K A, Wentzel E A, Zeller K I, et al. C-Myc-regulated microRNAs modulate E2F1 expression[J]. Nature, 2005,435 (7043):839-43.
[17] He L, Thomson J., Hemann M,et al. A microRNA polycistron as a potential human Oncogene[J]. Nature, 2005,435:828-833.
[18] Sylvestre Y, De Guire V, Querido E, et al. An E2F/miR-20a autoregulatory feedback loop[J]. J Biol Chem, 2007,282 (4): 2135-43.
[19] Tazawa H, Tsuchiya N, Izumiya M, et al. Tumor-suppressive miR-34a induces senescence-like growth arrest through modulation of the E2F pathway in human colon cancer cells[J]. Proc Natl Acad Sci USA,2007,104(39):15472-7.
[20] Yu F Y, Yao H, Zhu P C, et al. Let-7 Regulates self Renewal and Tumorigenicity of Breast Cancer Cells[J]. Cell,2007,12 (6): 1109 -1123.
[21] Hilah G, Gopal P, Andrew A., et al. MIR-451 and Imatinib mesylate inhibit tumor growth of Glioblastoma stem cells[J]. Biochemical and Biophysical Research Communications,2008,8 (31):86–9.
[22] Silber J,Lim D,Petritsch C,et al. MiR-124 and miR-137 inhibit proliferation of glioblastoma multiforme cells and induce differentiation of brain tumor stem cells[J]. BMC Medicine, 2008,6:14.
[23] Neumuller R, Betschinger J, Fischer A,et al. Mei-P26 regulates microRNAs and cell growth in the Drosophila ovarian stem cell lineage[J]. Nature,2008, 454(7201):241-54.
[24]黎丹戎,张汉英,张玮等.汉黄芩素对人卵巢癌细胞株SKOV3裸鼠移植瘤生长及端粒酶活性的抑制作用[J].中国药理学通报, 2007,23(4):119-123.
[24] LI D R,ZHANG H Y,ZHANG W, et al. The inhibitory role of wogonin on tumor growth and telomerase activity of human ovarian cancer SKOV3 cell line xenograf in nude mice[J]. Chinese Pharmacological Bulletin,2007, 23(4): 119-123.
[25]林红英,殷润婷,奚涛等.在肿瘤研究中转基因小鼠模型的研究进展[J].中国药理学通报,2007,23(1):4-8.
[25] LIN H Y, YIN R T,XI T ,et al. Transgenic mouse models of inhibition of tumor angiogenesis[J]. Chinese Pharmacological Bulletin, 2007,23(1):4-8.
[26] Liang Z X , Wu H, Santosh Reddy , et al. Blockade of invasion and metastasis of breast cancer cells via targeting CXCR4 with an artificial microRNA[J]. Biochemical and Biophysical Research Communications, 2007,363:542–546.