AG490对人宫颈癌Hela细胞增殖、凋亡及Bcl-2表达的影响
摘要
目的:观察JAK激酶抑制剂AG490对人宫颈癌细胞株Hela细胞增殖、凋亡及Bcl-2表达的影响。
方法:(1)培养人宫颈癌细胞系Hela细胞并在光学显微镜下观察活细胞生长情况并摄片。(2)Hela细胞在含有10μmol/l、20μmol/l、40μmol/l、80μmol/l的AG490中培养24h、48h、72h,应用光学显微镜动态观察细胞形态学变化;(3)通过四甲基偶氮唑蓝(MTT)比色法观察AG490对细胞增殖活力的影响,计算细胞生长抑制率和IC_(50)。(4)采用流式细胞仪(FCM)定量检测终浓度为20μmol/l、40μmol/l、80μmol/l的AG490作用Hela细胞48h后不同浓度组细胞的凋亡率,并分析AG490对细胞周期分布的影响。(5)采用终浓度为20μmol/l、40μmol/l、80μmol/l的AG490处理Hela48h,之后应用免疫细胞化学染色技术观察Hela细胞的凋亡相关基因Bcl-2表达的变化。各实验对照组均加入同等浓度的DMSO,未加入其他干预因素。
结果:(1)AG490以时间—剂量依赖性方式显著地抑制人宫颈癌细胞系Hela细胞体外的增殖活性(p＜0.05)。(2)经终浓度分别为10μmol/l、20μmol/l、40μmol/lAG490处理Hela细胞36h后,细胞被阻滞于G1期,S期细胞比例(SFP)明显降低(p＜0.05)。AG490作用24小时后,细胞bcl-2蛋白的表达降低,并出现典型的细胞凋亡的形态学表现,作用时间到48小时,Hela细胞凋亡达高峰。
结论:1.JAK激酶抑制剂AG490能有效抑制人宫颈癌Hela细胞的体外增殖,其作用呈时间—量效关系。
2.AG490下调Bcl-2蛋白表达,促进Hela细胞凋亡,凋亡率呈浓度依赖性。
Objective: To investigate the effect and its mechanisms of AG490 on the proliferation, apoptosis and Bcl-2 expression of human cervical carcinoma cell lines Hela cells in vitro.
Methods: Human cervical carcinoma cell lines Hela cells were cultured with different concentration of AG490 from 24 to 72 hours. The light microscopy was used to observe the changes morphologies of Hela cells.Methyl thiozolyl tetrazolium(MTT) was used to observe the growth suppressive rate of Hela. The flow cytometry (FCM) was used to investigate the phase of the proliferative cycles of Hela cells. The expression of Bcl-2 genes related to apoptosis were determinded by immunohistochemistry.
Results: The proliferation of Hela cells were suppressed by AG490 in time-dose dependent fashion in vitro(p<0.05).Hela cells were blocked at Glphase,and the S-phase fraction(SPF) of the cells were decreased. When the concentration of AG490 was increased to 20umol/l for 24 hours,the expression of bcl-2 proteins decreased and the typical morphological changes of apoptosis appeared at 48 hours by AG490 treatment.
方法:(1)培养人宫颈癌细胞系Hela细胞并在光学显微镜下观察活细胞生长情况并摄片。(2)Hela细胞在含有10μmol/l、20μmol/l、40μmol/l、80μmol/l的AG490中培养24h、48h、72h,应用光学显微镜动态观察细胞形态学变化;(3)通过四甲基偶氮唑蓝(MTT)比色法观察AG490对细胞增殖活力的影响,计算细胞生长抑制率和IC_(50)。(4)采用流式细胞仪(FCM)定量检测终浓度为20μmol/l、40μmol/l、80μmol/l的AG490作用Hela细胞48h后不同浓度组细胞的凋亡率,并分析AG490对细胞周期分布的影响。(5)采用终浓度为20μmol/l、40μmol/l、80μmol/l的AG490处理Hela48h,之后应用免疫细胞化学染色技术观察Hela细胞的凋亡相关基因Bcl-2表达的变化。各实验对照组均加入同等浓度的DMSO,未加入其他干预因素。
结果:(1)AG490以时间—剂量依赖性方式显著地抑制人宫颈癌细胞系Hela细胞体外的增殖活性(p＜0.05)。(2)经终浓度分别为10μmol/l、20μmol/l、40μmol/lAG490处理Hela细胞36h后,细胞被阻滞于G1期,S期细胞比例(SFP)明显降低(p＜0.05)。AG490作用24小时后,细胞bcl-2蛋白的表达降低,并出现典型的细胞凋亡的形态学表现,作用时间到48小时,Hela细胞凋亡达高峰。
结论:1.JAK激酶抑制剂AG490能有效抑制人宫颈癌Hela细胞的体外增殖,其作用呈时间—量效关系。
2.AG490下调Bcl-2蛋白表达,促进Hela细胞凋亡,凋亡率呈浓度依赖性。
Objective: To investigate the effect and its mechanisms of AG490 on the proliferation, apoptosis and Bcl-2 expression of human cervical carcinoma cell lines Hela cells in vitro.
Methods: Human cervical carcinoma cell lines Hela cells were cultured with different concentration of AG490 from 24 to 72 hours. The light microscopy was used to observe the changes morphologies of Hela cells.Methyl thiozolyl tetrazolium(MTT) was used to observe the growth suppressive rate of Hela. The flow cytometry (FCM) was used to investigate the phase of the proliferative cycles of Hela cells. The expression of Bcl-2 genes related to apoptosis were determinded by immunohistochemistry.
Results: The proliferation of Hela cells were suppressed by AG490 in time-dose dependent fashion in vitro(p<0.05).Hela cells were blocked at Glphase,and the S-phase fraction(SPF) of the cells were decreased. When the concentration of AG490 was increased to 20umol/l for 24 hours,the expression of bcl-2 proteins decreased and the typical morphological changes of apoptosis appeared at 48 hours by AG490 treatment.
引文
[1]Schindle C,Shual K,Prezioso VR,et al.Interferon-dependent tyrosine phosphorylation of a latent cytoplasmic transcription factors.Science.1992,5071:809-813.
[2]宋伦.JAK/STAT信号转导途径研究新进展.免疫学杂志,2000,16:68-71.
[3]Hsiehf C,Cheng G;Lin J,et al.Evaluation of potential Stat3-regulated genes in human breast cancer.Biochem Biophys Res Commun,2005,335(2):292-299.
[4]Lin Q,Lai R,Chiriealc LR,et al.Constitutive activation ofJAK3/STAT3in colon carcinoma tumors and cell lines:Inhibition of JAK3/STAT3 signaling induces apoptosis and cell cycle arrest of colon carcinoma cells.Am J Pathol.2005,167(4):969-980.
[5]Huang C,Cao J,Zhang F,et al.Inhibition of STAT3 Activity with AG490decreases the invasion of human pancreatic cancer cells in vitro.Cancer Sci,2006,97(12):1417-1423.
[6]Dagvadorj A,Collins S,Jomain JB,et al.Autocrine prolactin promotes prostate cancer cell growth via Janus kinase-2 signal transducer and activator of transcription-5a/b signaling pathway.Endocrinology,2007,148(7):3089-3101.
[7]Lacour S,Micheau Q,Hammann A,Drouineaud V,et al.Chemotherapy enhances TNF-related apoptosis-inducing ligand DISC assembly in HT29 human colon cells.Oncogene,2003,22:1807-1816.
[8]Ferrajoli A,Faderl S,Van Q,et al.WP1066 disrupts Janus kinase-2 and induces caspase-dependent apoptosis in acute myelogenous leukemia cells.Cancer Res.2007,67(23):11291-11299.
[9]Catlett-Falcone R,Landowski TH,Oshiro MM,et al.Constitutive activation of Stat3 singnaling confers resistance to apoptosis in human U266 myeloma cell.Immunity,1999,10(1):105-115.
[10]Yang SF,Yuan SS,YehYT,et al.Positive association between STAT3 and Ki67 in cervical intraepithelial neoplasia.Kaohsiung JMed Sci,2006,22(11):539-546.
[11] Chen CL, Hsieh FC, Lieblein JC,et al. Stat3 activation in human endometrial and cervical cancers. Br J Cancer, 2007,96 (4):591-599.
[12] Rosen DG, Mercado UI, Yang G, et al. The role of constitutive active signal transducer and activator of transcription3 in ovarian tumorigenesis and prognosis. Cancer, 2006,107(11):2730-2740.
[13]SilverD L, Naora H, Liu J, et al. Activated signal transducer and activator of transcription (STAT3) : Localization in focal adhesion and function in ovarian cancer cell molitity. Cancer Res, 2004,64(10):3550-3558.
[14] Moral B, Buettner R, Seigne J, et al. Constitutive activation of Stat3 in human prostate tumors and cell lines: direct inhibition of Stat3 signaling induces apoptosis of prostate cancer cells. Cancer Res, 2002,62(22):6659-6666.
[15] Eric BH, Zheng Z, Song LX, et al. Activated epidermal growth factor receptor-Stat3 signaling promotes tumor survival in vivo in non-small cell lung cancer. Clin Cancer Res, 2005,11 (23):8288-8294.
[16] Song L, Turkson J, Karras JG, et al. Activation of STAT3 by receptor tyrosine kinases and cytokines regulates survival in human non—small cell carcinoma cells. Oncogene, 2003,22(27):4150-4165.
[17] Miyamoto N, Sugita K, Goi K, et al. The JAK2 inhibitor AG490 predominantly abrogates the growth of human B-precursor leukemic cells with 11q23 translocation or Philadelphia chromosome. Leukemia, 2001,15:1758-1768.
[18]Burdelya L, Catlett-Falcone R, Levitzki A, et al. Combination therapy with AG490 and interleukin 12 achieves greater antitumor effects than either agent alone. Mol Cancer Ther, 2002,1:893-899.
[19]Gesbert F , Griffin JD. Bcr/Abl activates transcription of the bcl-x genes through STAT5. Blood, 2000,96:2269-2267.
[20]Epling-Burnette PK, Liu JH, Catlett-Falcone R, et al. Inhibition of STAT3 signaling leads to apoptosis of Leukemic large granular lymphocytes and decreasesd Mcl-1 expression. J Clin Investing, 2001,107:351-362.
[21] Bowman T, Broome MA, Sinibaldi D, et al. Stat3-mediated Myc expression is required for Src transformation and PDGF-induced mitogenesis. Proc Natl Acad Sci,2001,98:7319-7324.
[22]Matsui T,Kinoshita T,Hirano T,et al.STAT3 down-regulates the expression of cyclin D during liver development.J Biol Chem,2002,277:36167-36173.
[23]Hodge DR,Hurt EM,arrar WL.The role of IL-6 and STAT3 in inflammation and cancer.Eur J Cancer,2005,41(16):2502-2512.
[24]Yu H,Jove R.The STATs of cancer-new molecular targets come of age.Nat Rev Cancer,2004,4(2):97-105.
[25]Buettner R,Mora LB,Jove R.Activated STAT signaling in human tumors provides novel molecular targets for therapeutic intervention.Clin Cancer Res,2002,8(4):945-954.
[26]Toyonaga T,Nakano K,Nagano M,et al.Blockade of constitutively activated Janus kinase/signal transducer and activator of transcription pathway inhibits growth of human pancreatic cancer.Cancer Lett,2003,201(1):107-116.
[27]黄陈,裘正军,刘辰,等.阻断STAT3信号转导通路对人胰腺癌细胞生长的影响.肿瘤,2006,26(5):414-417.
[28]Marico T,Hirochika K,Jun-nosuke U,et al.Inhibition of constitutively active Jak-stat pathway suppression cell growth of human T-cell leukemia virus type-infected T-cell lines and primary adult T-cell leukemia celia.Retrovirology,2006,3(22):1742-1746.
[29]司徒镇强,吴军正,等.细胞培养(第1版).世界图书出版公司.2004,3:197.
[30]Fuke H,Shirakik K,Suqimoto K,et al.Jak inhibitor induces S phase cell-cycle arrest and augments TRAIL-induced apoptosis in human hepatocellular carcinoma cells.Biochem Biophys Res Commun,2007,363(3):738-744.
[31]Iwamaru A,Szymanski S,Iwado S,et al.A novel inhibitor of the stst3 pathway induces apoptosis in malignant glioma cellsboth in vitro and in vivo.Oncogene,2007,26(17):2435-2444.
[32]黎明红,姚震.妇科恶性肿瘤.医学综述.2002,8(1):5-7.
[33]Ferrandina G,Moazztti S,Marone M,et al.bcl-2,bax,bcl-x(L)and bcl-x(S)expression in neoplastic and normal cervical tissue.Cancer Lett,2000,155(1):19-27.
[34]Ozalp SS, Yalcin OT, Tanir HM, et al. Bcl-2 expression in Preinvasive and in vasive cervical lesions. Eur J Gynaecol Oncol. 2002,23(5):419-422.
[35]Shao YF, Gao ZH, Paul A, et al. Apoptosis and autophagic cell death induced by histone deacetylase inhibitors. Cell Biology. 2004,101(52): 18030-18035.
[36]Xiong H, Zhang ZG, Tian XQ, et al. Inhibition of Jak1,2/stat3 signaling induces apoptosis, cell cycle arrest, and reduced tumor cell invasion in Colorectal cancer cell. Neoplasia, 2008,10(3):287-297.
[37] Steve A, Benjamin B, et al. Inhibition of constitutive STAT3 activity sentisizes resistant non-hodgkin's lymphoma and multiple myeloma to chemotherapeutic drug-mediated apoptosis. Clinical Cancer Res. 2003,9:316-326.
[1] Krebs DL, Hilton DJ. SOCS proteins: Negative regulators of cytokine signaling. Stem Cells, 2001,19(5):378-387.
[2] Copeland NG, Gilbert DJ, Schindler C, et al. Distribution of the mammalian Stat gene family in mouse chromosomes. Genomics. 1995,29:225-228.
[3] Hsiehf C, Cheng G, Lin J, et al. Evaluation of potential Stat3-regulated genes in human breast cancer. Biochem Biophys Res Commun, 2005,335(2):292-299.
[4] Lin Q, Lai R, Chiriealc LR, et al. Constitutive activation of JAK3/STAT3 in colon carcinoma tumors and cell lines: Inhibition of JAK3/STAT3 signaling induces apoptosis and cell cycle arrest of colon carcinoma cells. Am J Pathol. 2005, 167(4): 969-980.
[5] Huang C, Cao J, Zhang F, et al. Inhibition of STAT3 Activity with AG490 decreases the invasion of human pancreatic cancer cells in vitro. Cancer Sci, 2006,97(12):1417-1423.
[6] Dagvadorj A, Collins S, Jomain JB, et al. Autocrine prolactin promotes prostate cancer cell growth via Janus kinase-2-signal transducer and activator of transcription-5a/b signaling pathway. Endocrinology, 2007,148(7):3089-3101.
[7] Lacour S, Micheau Q, Hammann A, Drouineaud V, et al. Chemotherapy enhances TNF-related apoptosis-inducing ligand DISC assembly in HT29 human colon cells. Oncogene, 2003,22:1807-1816.
[8] Ferrajoli A, Faderl S, Van Q, et al. WP1066 disrupts Janus kinase-2 and induces caspase-dependent apoptosis in acute myelogenous leukemia cells. Cancer Res. 2007,67(23):11291-11299.
[9] Chen CL, Hsieh FC, Lieblein JC, et al. Stat3 activation in human endometrial and cervical cancers. Br J Cancer, 2007,96 (4):591-599.
[10]Yang SF, Yuan SS, Yeh Y T, et al. Positive association between STAT3 and Ki67 in cervical intraepithelial neoplasia. Kaohsiung JMed Sc, 2006,22(11):539-546.
[11] Hodge DR, Hurt EM, Farrar WL, et al. The role of IL-6 and STAT3 in inflammation and cancer. Eur J Cancer, 2005,41 (16):2502-2512.
[12] Yu H, Jove R. The STATs of cancer-new molecular targets come of age. Nat Rev Cancer, 2004,4(2): 97-105.
[13]Buettner R, Mora LB, Jove R. Activated STAT signaling in human tumors provides novel molecular targets for therapeutic intervention. Clin Cancer Res, 2002,8(4):945-954.
[14]Korty L, Yu H, et al. Stat3 as a potential target for cancer immunotherapy, Journal of immunotherapy, 2007,30(2):131-139.
[15]Krebs DL, Hilton DJ. SOCS proteins: Negative regulalors of cytokine signaling. Stem Cells, 2001,19 (5): 378-387.
[16]Yamamura Y, Senda H, Noda M, et al. Leukemia, 1997,11(Suppl 3):432.
[17]Danial NN, Osman JA, Lu T, et al. Mol Cell Biol, 1998,18:67-95.
[18]Migone TS, Lin JX, Cereseto A, et al. Science, 1995,269-279.
[19]Niesen M, kaestel CG, Eriksen KW, et al. Leukemia, 1999,13:735.
[20] Miyamoto N, Sugita K, Goi K, et al. The JAK2 inhibitor AG490 predominantly abrogates the growth of human B-precursor leukemic cells with 11q23 translocation or Philadelphia chromosome. Leukemia, 2001,15:1758-1768.
[21]Thyrell L, Hjortsberg L, Arulampalam V, et al. Inter-feron alpha-induced apoptosis in tumor cells ismediated through the phosphoinositide 3-kinase/mammalian target of rapamycin signaling pathway. J Biol Chem, 2004, 279: 24152-24162.
[22]Kisseleva T, Bhattacharya S, Braunstein J, et al. Signaling through the JAK/STAT pathway, recent advances and future challenges. Gene, 2002,285:12-24.
[23]Kochendoerfer SK, Krishnan N, Buckley DJ, et al. Prolactin regulation of Bcl-2 family members: increased expression of Bcl-xL but not mcl-1 or bad in Nb2-T cells. J Endocrinol, 2003,178:265-273.
[24] Zhang X, Zhang H, Ye L. Effects of hepatitis B virus X protein on development of liver cancer. J Lab Clin Med, 2006,147(2):58-66.
[25]Waris G, Huh KW, and Siddiqui A. Mitochondrially Associated Hepatitis B Virus X protein constitutively activates transcription factors STAT3 and NF-κb via Oxidative stress.Molecular and cellular biology,2001,21(22):7721-7730.
[26]Waris G,Siddiqui A.Regulatory mechanisms of viral hepatitis B and C.J Biosci,2003,28(3):311-321.
[27]Yoshikawa H,Matsubara K,Qian GS,et al.SOCS-1,a negative regulator of the JAK/STAT pathway is silenced by methylation in human hepatocellular carcinoma and shows growth suppression activity.Nat Genet,2001,28(1):29-35.
[28]Toyonaga T,Nakano K,Nagano M,et al.Blockade of constitutively activated Janus kinase/signal transducer and activator of transcription pathway inhibits growth of human pancreatic cancer.Cancer Lett,2003,201(1):107-116.
[29]Huang C,Cao J,Zhang F,et al.Inhibition of STAT3 Activity with AG490decreases the invasion of human pancreatic cancer ceils in vitro.Cancer Sci,2006,97(12):1417-1423.
[30]姚林方,叶章群,陈志强,等.信号传导阻滞剂AG490抑制膀胱癌细胞生长的研究.肿瘤防治研究.2007,34(3):195-198.
[31]Kovanen PE,Junttila I,Takaluoma K,et al.Regulation of JAK2 tyrosine kinase by protein kinase C during macrophage differentiation of IL-3-dependeng myeloid progenitor cells.Blood,2000,95(5):1626-1632.
[32]董燕,梅柱中,钱俊杰,等.人外周血活化淋巴细胞中Survivin蛋白表达的分子机制研究.细胞与分子免疫学杂志,2008,24(1):16-19.
[33]王彬,冯新顺,薛武军,等.AG490对人淋巴细胞增殖过程中细胞因子分泌的作用.中华器官移植杂志,2005,26(2):72-74.
[34]王艳春,黄进明,朱明光,等.AG490阻断肝癌细胞Stat3信号通路逆转免疫抑制的实验研究.细胞与分子免疫学杂志,2007,23(12):1170-1171.
[35]Stroll G,Jander S,Schroeter M.Determental and beneficial effects of injury-induced inflammation and cytokine expression in the nervous system.Adv Exp Med Biol,2002,513:87-113.
[36]Iadecola C,Alexander M.Cerebral ischemic and inflammation.Curr Opin Neurol,2001,14(1):89-94.
[37]Yamashita T,Sawamoto K,Suzuki S,et al.Blockade of interleukin-6 signaling aggravates ischemic cerebral damage in mice:possible involvement of STAT3activation in the protection of neurons.J Neurochem,2005,94(2):459-468.
[38]Satriotomo I,Bowen kk,Vemuganti R.JAK2 and STAT3 activation contributes to neuronal damage following transient focal cerebral ischemia.J Neurochem,2006,98(5):1353-1368.
[39]Smith RM,Suleman N,Lacerda L,et al.Genetic depletion of cardiac myocyte STAT-3 abolishes classical preconditioning.Cardiovasc Res,2004,63:611-616.
[40]Yamaura G,Turoczi T,Yamamoto F,et al.STAT signaling in ischemic heart:a role of STAT5A in ischemic preconditioning.Am J Physiol Heart Circ Physiol,2003,5:476-482.
[41]Mascareno E,El- Shafei M,Maulik N,et al.JAK/STAT signaling is associated with cardiac dysfunction during ischemia and reperfusion.Circulation,2001,104:325-329.
[42]Hattori R,Maulik N,Otani H,et al.Role of STAT3 in ischemic preconditioning.J Mol Cell Cardiol,2001,33:1929-1936.
[43]唐梅,张钲,伏静媛,JAK2/STAT3信号通路介导大鼠血管平滑肌细胞的增殖与凋亡.基础医学与临床,2006,2:1354-1358.
[44]王松柏,姚咏明,等.Janus激酶/信号传导和转录激活因子途径与脓毒症的研究进展.中国危重病急救医学,2003,15:690-693.
[45]王松柏,翟秀珍,姚咏明,等.AG490预先给药对脓毒血症大鼠血清和肾组织肿瘤坏死因子-a水平的影响.中华麻醉学杂志,2005,25(9):699-700.
[46]姚胜,姚咏明,李红云,等.抑制JA K/STAT通路对烫伤后金黄色葡萄球菌脓毒症大鼠肝损害的影响.中国危重病急救医学,2002,14(6):336-339.
[2]宋伦.JAK/STAT信号转导途径研究新进展.免疫学杂志,2000,16:68-71.
[3]Hsiehf C,Cheng G;Lin J,et al.Evaluation of potential Stat3-regulated genes in human breast cancer.Biochem Biophys Res Commun,2005,335(2):292-299.
[4]Lin Q,Lai R,Chiriealc LR,et al.Constitutive activation ofJAK3/STAT3in colon carcinoma tumors and cell lines:Inhibition of JAK3/STAT3 signaling induces apoptosis and cell cycle arrest of colon carcinoma cells.Am J Pathol.2005,167(4):969-980.
[5]Huang C,Cao J,Zhang F,et al.Inhibition of STAT3 Activity with AG490decreases the invasion of human pancreatic cancer cells in vitro.Cancer Sci,2006,97(12):1417-1423.
[6]Dagvadorj A,Collins S,Jomain JB,et al.Autocrine prolactin promotes prostate cancer cell growth via Janus kinase-2 signal transducer and activator of transcription-5a/b signaling pathway.Endocrinology,2007,148(7):3089-3101.
[7]Lacour S,Micheau Q,Hammann A,Drouineaud V,et al.Chemotherapy enhances TNF-related apoptosis-inducing ligand DISC assembly in HT29 human colon cells.Oncogene,2003,22:1807-1816.
[8]Ferrajoli A,Faderl S,Van Q,et al.WP1066 disrupts Janus kinase-2 and induces caspase-dependent apoptosis in acute myelogenous leukemia cells.Cancer Res.2007,67(23):11291-11299.
[9]Catlett-Falcone R,Landowski TH,Oshiro MM,et al.Constitutive activation of Stat3 singnaling confers resistance to apoptosis in human U266 myeloma cell.Immunity,1999,10(1):105-115.
[10]Yang SF,Yuan SS,YehYT,et al.Positive association between STAT3 and Ki67 in cervical intraepithelial neoplasia.Kaohsiung JMed Sci,2006,22(11):539-546.
[11] Chen CL, Hsieh FC, Lieblein JC,et al. Stat3 activation in human endometrial and cervical cancers. Br J Cancer, 2007,96 (4):591-599.
[12] Rosen DG, Mercado UI, Yang G, et al. The role of constitutive active signal transducer and activator of transcription3 in ovarian tumorigenesis and prognosis. Cancer, 2006,107(11):2730-2740.
[13]SilverD L, Naora H, Liu J, et al. Activated signal transducer and activator of transcription (STAT3) : Localization in focal adhesion and function in ovarian cancer cell molitity. Cancer Res, 2004,64(10):3550-3558.
[14] Moral B, Buettner R, Seigne J, et al. Constitutive activation of Stat3 in human prostate tumors and cell lines: direct inhibition of Stat3 signaling induces apoptosis of prostate cancer cells. Cancer Res, 2002,62(22):6659-6666.
[15] Eric BH, Zheng Z, Song LX, et al. Activated epidermal growth factor receptor-Stat3 signaling promotes tumor survival in vivo in non-small cell lung cancer. Clin Cancer Res, 2005,11 (23):8288-8294.
[16] Song L, Turkson J, Karras JG, et al. Activation of STAT3 by receptor tyrosine kinases and cytokines regulates survival in human non—small cell carcinoma cells. Oncogene, 2003,22(27):4150-4165.
[17] Miyamoto N, Sugita K, Goi K, et al. The JAK2 inhibitor AG490 predominantly abrogates the growth of human B-precursor leukemic cells with 11q23 translocation or Philadelphia chromosome. Leukemia, 2001,15:1758-1768.
[18]Burdelya L, Catlett-Falcone R, Levitzki A, et al. Combination therapy with AG490 and interleukin 12 achieves greater antitumor effects than either agent alone. Mol Cancer Ther, 2002,1:893-899.
[19]Gesbert F , Griffin JD. Bcr/Abl activates transcription of the bcl-x genes through STAT5. Blood, 2000,96:2269-2267.
[20]Epling-Burnette PK, Liu JH, Catlett-Falcone R, et al. Inhibition of STAT3 signaling leads to apoptosis of Leukemic large granular lymphocytes and decreasesd Mcl-1 expression. J Clin Investing, 2001,107:351-362.
[21] Bowman T, Broome MA, Sinibaldi D, et al. Stat3-mediated Myc expression is required for Src transformation and PDGF-induced mitogenesis. Proc Natl Acad Sci,2001,98:7319-7324.
[22]Matsui T,Kinoshita T,Hirano T,et al.STAT3 down-regulates the expression of cyclin D during liver development.J Biol Chem,2002,277:36167-36173.
[23]Hodge DR,Hurt EM,arrar WL.The role of IL-6 and STAT3 in inflammation and cancer.Eur J Cancer,2005,41(16):2502-2512.
[24]Yu H,Jove R.The STATs of cancer-new molecular targets come of age.Nat Rev Cancer,2004,4(2):97-105.
[25]Buettner R,Mora LB,Jove R.Activated STAT signaling in human tumors provides novel molecular targets for therapeutic intervention.Clin Cancer Res,2002,8(4):945-954.
[26]Toyonaga T,Nakano K,Nagano M,et al.Blockade of constitutively activated Janus kinase/signal transducer and activator of transcription pathway inhibits growth of human pancreatic cancer.Cancer Lett,2003,201(1):107-116.
[27]黄陈,裘正军,刘辰,等.阻断STAT3信号转导通路对人胰腺癌细胞生长的影响.肿瘤,2006,26(5):414-417.
[28]Marico T,Hirochika K,Jun-nosuke U,et al.Inhibition of constitutively active Jak-stat pathway suppression cell growth of human T-cell leukemia virus type-infected T-cell lines and primary adult T-cell leukemia celia.Retrovirology,2006,3(22):1742-1746.
[29]司徒镇强,吴军正,等.细胞培养(第1版).世界图书出版公司.2004,3:197.
[30]Fuke H,Shirakik K,Suqimoto K,et al.Jak inhibitor induces S phase cell-cycle arrest and augments TRAIL-induced apoptosis in human hepatocellular carcinoma cells.Biochem Biophys Res Commun,2007,363(3):738-744.
[31]Iwamaru A,Szymanski S,Iwado S,et al.A novel inhibitor of the stst3 pathway induces apoptosis in malignant glioma cellsboth in vitro and in vivo.Oncogene,2007,26(17):2435-2444.
[32]黎明红,姚震.妇科恶性肿瘤.医学综述.2002,8(1):5-7.
[33]Ferrandina G,Moazztti S,Marone M,et al.bcl-2,bax,bcl-x(L)and bcl-x(S)expression in neoplastic and normal cervical tissue.Cancer Lett,2000,155(1):19-27.
[34]Ozalp SS, Yalcin OT, Tanir HM, et al. Bcl-2 expression in Preinvasive and in vasive cervical lesions. Eur J Gynaecol Oncol. 2002,23(5):419-422.
[35]Shao YF, Gao ZH, Paul A, et al. Apoptosis and autophagic cell death induced by histone deacetylase inhibitors. Cell Biology. 2004,101(52): 18030-18035.
[36]Xiong H, Zhang ZG, Tian XQ, et al. Inhibition of Jak1,2/stat3 signaling induces apoptosis, cell cycle arrest, and reduced tumor cell invasion in Colorectal cancer cell. Neoplasia, 2008,10(3):287-297.
[37] Steve A, Benjamin B, et al. Inhibition of constitutive STAT3 activity sentisizes resistant non-hodgkin's lymphoma and multiple myeloma to chemotherapeutic drug-mediated apoptosis. Clinical Cancer Res. 2003,9:316-326.
[1] Krebs DL, Hilton DJ. SOCS proteins: Negative regulators of cytokine signaling. Stem Cells, 2001,19(5):378-387.
[2] Copeland NG, Gilbert DJ, Schindler C, et al. Distribution of the mammalian Stat gene family in mouse chromosomes. Genomics. 1995,29:225-228.
[3] Hsiehf C, Cheng G, Lin J, et al. Evaluation of potential Stat3-regulated genes in human breast cancer. Biochem Biophys Res Commun, 2005,335(2):292-299.
[4] Lin Q, Lai R, Chiriealc LR, et al. Constitutive activation of JAK3/STAT3 in colon carcinoma tumors and cell lines: Inhibition of JAK3/STAT3 signaling induces apoptosis and cell cycle arrest of colon carcinoma cells. Am J Pathol. 2005, 167(4): 969-980.
[5] Huang C, Cao J, Zhang F, et al. Inhibition of STAT3 Activity with AG490 decreases the invasion of human pancreatic cancer cells in vitro. Cancer Sci, 2006,97(12):1417-1423.
[6] Dagvadorj A, Collins S, Jomain JB, et al. Autocrine prolactin promotes prostate cancer cell growth via Janus kinase-2-signal transducer and activator of transcription-5a/b signaling pathway. Endocrinology, 2007,148(7):3089-3101.
[7] Lacour S, Micheau Q, Hammann A, Drouineaud V, et al. Chemotherapy enhances TNF-related apoptosis-inducing ligand DISC assembly in HT29 human colon cells. Oncogene, 2003,22:1807-1816.
[8] Ferrajoli A, Faderl S, Van Q, et al. WP1066 disrupts Janus kinase-2 and induces caspase-dependent apoptosis in acute myelogenous leukemia cells. Cancer Res. 2007,67(23):11291-11299.
[9] Chen CL, Hsieh FC, Lieblein JC, et al. Stat3 activation in human endometrial and cervical cancers. Br J Cancer, 2007,96 (4):591-599.
[10]Yang SF, Yuan SS, Yeh Y T, et al. Positive association between STAT3 and Ki67 in cervical intraepithelial neoplasia. Kaohsiung JMed Sc, 2006,22(11):539-546.
[11] Hodge DR, Hurt EM, Farrar WL, et al. The role of IL-6 and STAT3 in inflammation and cancer. Eur J Cancer, 2005,41 (16):2502-2512.
[12] Yu H, Jove R. The STATs of cancer-new molecular targets come of age. Nat Rev Cancer, 2004,4(2): 97-105.
[13]Buettner R, Mora LB, Jove R. Activated STAT signaling in human tumors provides novel molecular targets for therapeutic intervention. Clin Cancer Res, 2002,8(4):945-954.
[14]Korty L, Yu H, et al. Stat3 as a potential target for cancer immunotherapy, Journal of immunotherapy, 2007,30(2):131-139.
[15]Krebs DL, Hilton DJ. SOCS proteins: Negative regulalors of cytokine signaling. Stem Cells, 2001,19 (5): 378-387.
[16]Yamamura Y, Senda H, Noda M, et al. Leukemia, 1997,11(Suppl 3):432.
[17]Danial NN, Osman JA, Lu T, et al. Mol Cell Biol, 1998,18:67-95.
[18]Migone TS, Lin JX, Cereseto A, et al. Science, 1995,269-279.
[19]Niesen M, kaestel CG, Eriksen KW, et al. Leukemia, 1999,13:735.
[20] Miyamoto N, Sugita K, Goi K, et al. The JAK2 inhibitor AG490 predominantly abrogates the growth of human B-precursor leukemic cells with 11q23 translocation or Philadelphia chromosome. Leukemia, 2001,15:1758-1768.
[21]Thyrell L, Hjortsberg L, Arulampalam V, et al. Inter-feron alpha-induced apoptosis in tumor cells ismediated through the phosphoinositide 3-kinase/mammalian target of rapamycin signaling pathway. J Biol Chem, 2004, 279: 24152-24162.
[22]Kisseleva T, Bhattacharya S, Braunstein J, et al. Signaling through the JAK/STAT pathway, recent advances and future challenges. Gene, 2002,285:12-24.
[23]Kochendoerfer SK, Krishnan N, Buckley DJ, et al. Prolactin regulation of Bcl-2 family members: increased expression of Bcl-xL but not mcl-1 or bad in Nb2-T cells. J Endocrinol, 2003,178:265-273.
[24] Zhang X, Zhang H, Ye L. Effects of hepatitis B virus X protein on development of liver cancer. J Lab Clin Med, 2006,147(2):58-66.
[25]Waris G, Huh KW, and Siddiqui A. Mitochondrially Associated Hepatitis B Virus X protein constitutively activates transcription factors STAT3 and NF-κb via Oxidative stress.Molecular and cellular biology,2001,21(22):7721-7730.
[26]Waris G,Siddiqui A.Regulatory mechanisms of viral hepatitis B and C.J Biosci,2003,28(3):311-321.
[27]Yoshikawa H,Matsubara K,Qian GS,et al.SOCS-1,a negative regulator of the JAK/STAT pathway is silenced by methylation in human hepatocellular carcinoma and shows growth suppression activity.Nat Genet,2001,28(1):29-35.
[28]Toyonaga T,Nakano K,Nagano M,et al.Blockade of constitutively activated Janus kinase/signal transducer and activator of transcription pathway inhibits growth of human pancreatic cancer.Cancer Lett,2003,201(1):107-116.
[29]Huang C,Cao J,Zhang F,et al.Inhibition of STAT3 Activity with AG490decreases the invasion of human pancreatic cancer ceils in vitro.Cancer Sci,2006,97(12):1417-1423.
[30]姚林方,叶章群,陈志强,等.信号传导阻滞剂AG490抑制膀胱癌细胞生长的研究.肿瘤防治研究.2007,34(3):195-198.
[31]Kovanen PE,Junttila I,Takaluoma K,et al.Regulation of JAK2 tyrosine kinase by protein kinase C during macrophage differentiation of IL-3-dependeng myeloid progenitor cells.Blood,2000,95(5):1626-1632.
[32]董燕,梅柱中,钱俊杰,等.人外周血活化淋巴细胞中Survivin蛋白表达的分子机制研究.细胞与分子免疫学杂志,2008,24(1):16-19.
[33]王彬,冯新顺,薛武军,等.AG490对人淋巴细胞增殖过程中细胞因子分泌的作用.中华器官移植杂志,2005,26(2):72-74.
[34]王艳春,黄进明,朱明光,等.AG490阻断肝癌细胞Stat3信号通路逆转免疫抑制的实验研究.细胞与分子免疫学杂志,2007,23(12):1170-1171.
[35]Stroll G,Jander S,Schroeter M.Determental and beneficial effects of injury-induced inflammation and cytokine expression in the nervous system.Adv Exp Med Biol,2002,513:87-113.
[36]Iadecola C,Alexander M.Cerebral ischemic and inflammation.Curr Opin Neurol,2001,14(1):89-94.
[37]Yamashita T,Sawamoto K,Suzuki S,et al.Blockade of interleukin-6 signaling aggravates ischemic cerebral damage in mice:possible involvement of STAT3activation in the protection of neurons.J Neurochem,2005,94(2):459-468.
[38]Satriotomo I,Bowen kk,Vemuganti R.JAK2 and STAT3 activation contributes to neuronal damage following transient focal cerebral ischemia.J Neurochem,2006,98(5):1353-1368.
[39]Smith RM,Suleman N,Lacerda L,et al.Genetic depletion of cardiac myocyte STAT-3 abolishes classical preconditioning.Cardiovasc Res,2004,63:611-616.
[40]Yamaura G,Turoczi T,Yamamoto F,et al.STAT signaling in ischemic heart:a role of STAT5A in ischemic preconditioning.Am J Physiol Heart Circ Physiol,2003,5:476-482.
[41]Mascareno E,El- Shafei M,Maulik N,et al.JAK/STAT signaling is associated with cardiac dysfunction during ischemia and reperfusion.Circulation,2001,104:325-329.
[42]Hattori R,Maulik N,Otani H,et al.Role of STAT3 in ischemic preconditioning.J Mol Cell Cardiol,2001,33:1929-1936.
[43]唐梅,张钲,伏静媛,JAK2/STAT3信号通路介导大鼠血管平滑肌细胞的增殖与凋亡.基础医学与临床,2006,2:1354-1358.
[44]王松柏,姚咏明,等.Janus激酶/信号传导和转录激活因子途径与脓毒症的研究进展.中国危重病急救医学,2003,15:690-693.
[45]王松柏,翟秀珍,姚咏明,等.AG490预先给药对脓毒血症大鼠血清和肾组织肿瘤坏死因子-a水平的影响.中华麻醉学杂志,2005,25(9):699-700.
[46]姚胜,姚咏明,李红云,等.抑制JA K/STAT通路对烫伤后金黄色葡萄球菌脓毒症大鼠肝损害的影响.中国危重病急救医学,2002,14(6):336-339.