紫杉醇抑制HepG2细胞生长的作用机理研究
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摘要
肝癌是一种发生于肝脏的恶性肿瘤,它包括原发性(原发部位在肝脏)与转移性(由其他部位转移而来)肝癌两种,一般的肝癌指的是原发性肝癌。原发性肝癌以细胞分型可分为胆管细胞型肝癌、肝细胞型肝癌和混合型肝癌。全球每年新发肝癌患者约六十万,居恶性肿瘤第五位。有关肝癌的治疗方法较多,但尚无有效的治疗,化学治疗被广泛应用于肝癌。
紫杉醇被应用于临床肿瘤治疗,它红豆杉属的植物中有一种具有抗癌作用十分明显的结构相当复杂次生代谢产物。紫杉醇能通过与微管蛋白相结合,阻止微管解聚,阻断了细胞的正常分裂,使细胞分裂停止。因此,阻滞了细胞周期的正常运转,抑制癌细胞生长,从而起到抗癌作用。然而,紫杉醇治疗癌症的分子机理还不十分清楚。
硫氧还蛋白结合蛋白-2(Thioredoxin Binding Protein-2,TBP-2)是一种广泛存在于人体各组织中的分子量为46kD蛋白质。维生素D3能够诱导TBP-2在细胞中的表达量增加,所以,TBP-2又名叫维生素D3上调蛋白1(vitamin D3up regulated protein-1,VDUP-1), TBP-2与还原型硫氧还蛋白结合从而抑制其活性。当硫氧还蛋白半胱氨酸活性位点发生突变时硫氧还蛋白结合蛋白-2便不能与之相结合。因此,TBP-2是硫氧还蛋白的一个负性调节蛋白。通过在一些癌细胞中导入外源TBP-2表达质粒使其表达量增加,则可抑制细胞生长,促进细胞凋亡。TBP-2与糖代谢密切相关,葡萄糖可以诱导TBP-2的表达。
硫氧还蛋白(thioredoxin,Trx)是一种广泛分部在各种细胞和各种组织中的分子量为12kDa的多功能蛋白分子,硫氧还蛋白大约含有110个氨基酸这些氨基酸中有一个功能活性区域-Cys-Gly-Pro-Cys-,硫氧还蛋白同硫氧还蛋白还原酶(Thioredoxin reductase, TrxR)烟酰胺腺嘌呤二核苷酸磷酸(NADPH)三者一起组成硫氧还蛋白还原系统,一起调节某些转录因子的活性,调节细胞中的氧化还原反应的平衡。硫氧还蛋白还能与凋亡信号调节激酶1(apoptosis signal-regulating kinase1, ASK1)相结合,抑制细胞凋亡的途径。硫氧还蛋白还具有促进肿瘤血管再生和加速细胞生长等作用,与缺氧诱导因子及细胞周期蛋白关系密切。
本课题将使用将使用高浓度葡萄糖及紫杉醇刺激HepG2细胞,观察细胞活性变化,并且观察高糖是否促进紫杉醇诱导HepG2细胞凋亡；并研究紫杉醇作用后硫氧还蛋白,TBP-2,低氧诱导因子-1α,以及cyclinD1的表达；并使用SiRNA降低TBP-2的表达,检测在TBP-2低表达的情况下HepG2细胞对紫杉醇的敏感性。
研究结果：紫杉醇降低HepG2细胞活性,但高浓度葡萄糖并不促进紫杉醇的作用。紫杉醇抑制硫氧还蛋白及cyclinD1的表达。紫杉醇对TBP-2的表达无影响。TBP-2低表达的情况下对紫杉醇的抗癌作用具有抵抗作用性。
以上结果表明：紫杉醇通过抑制硫氧还蛋白,低氧诱导因子-1α以及cyclinD1的表达降低HepG2的活性,抑制其生长,促进凋亡。紫杉醇不具有诱导TBP-2的作用,但TBP-2低表达可以部分抵抗紫杉醇的作用。因此,硫氧还蛋白和TBP-2均可以作为紫杉醇的一个新的作用靶点。
Liver cancer is the malignant tumor in the liver, including primary liver cancer and metastatic liver cancer, liver cancer refers to people' daily said more than primary liver cancer. Primary liver cancer is one of the most common malignancies in the clinical, Primary liver cancer cells can be divided into the hepatocellular carcinoma, cholangiocarcinoma and mixed tumors. According to the latest statistics, around the world every year about six hundred thousand new cases of liver cancer patients, ranking fifth in the malignant tumor. There are many treatment methods on liver cancer,however,the effective mothod is few, chemotherapy is a widely used in treatment of liver cancer.
Paclitaxel is used as tumors in clinical, is a complex secondary metabolites in the genus Taxus, and is currently understood only one that can promote microtubules polymerization and stability of the polymerized microtubule drugs. Isotope tracing showed that Paclitaxel only binds to the polymerization of microtubules, inhibits its depolymerization, blocking the normal cell division, especially makes cell division stopped in mitosis, blocks the cell cycle, inhibits the cell growth, then plays role in anticancer. However, its molecular mechanisms on treating cancer are still unknown.
Thioredoxin binding protein-2is also called vitamin D3up regulated protein-1, or thioredoxin interacting protein, the molecular weight is46kDa, it was first isolated in HL-60cells, vitamin D3can induce the expression. TBP-2is the Trx endogenous inhibitor of Trx activities. TBP-2binds with the reduced state of Trx only, not bind with the oxidation state or activity site cysteine mutant, therefore, TBP-2is an inhibitor of Trx. TBP-2were transfected into some cells, the cell growth was inhibited, the apoptosis occurred. TBP-2is closely relate with glycometabolism. Glucose could induce the expression of TBP-2.
Thioredoxin is a molecular weight of12kDa protein, it has a highly conservative active site that is-Cys-Gly-Pro-Cys-, which has a variety of biological functions. Trx, NADPH and Thioredoxin reductase composed thioredoxin system, adjusting the intracellular oxidation-reduction equilibrium, regulating the activity of transcription factors. At the same time, Trx also combine with apoptosis signal-regulating kinase1,(ASK1), and also can promote the growth of cells and blood vessels regeneration. It is also associated with hypoxia-inducible factor-la (HIF-la) and cyclinD1.
This study will use high concentration of glucose and paclitaxel treated HepG2cells and detected cell viability, and observed whether high concentration of glucose enhanced the susceptibility of Paclitaxel in HepG2cells and promoted apoptosis by Paclitaxel. To examine the expressions of Thioredoxin, TBP-2HIFla and cyclinDl induced by Paclitaxel. By using SiRNA to decrease TBP-2expression, at lower expression of TBP-2, detected the HepG2cells susceptibility to Paclitaxel.
Results:Paclitaxel decreased cell viability in HepG2, but high concentration of glucose did not enhance the role of Paclitaxel. Paclitaxel inhibited the expressions of thioredoxin, cyclinDl and HIF-1α but not TBP-2. Lower expression of TBP-2had resistance to Paclitaxel.
Above results showed:Paclitaxel decreased cell viability, inhibited cell growth and induce apoptosis in HepG2by inhibiting the expressions of Thioredoxin, HIF-la and cyclinDl. Paclitaxel did not induce the expression of TBP-2, but the Lower expression of TBP-2had resistance to Paclitaxel.Thus, thioredoxin and TBP-2is a new target of Paclitaxel.
紫杉醇被应用于临床肿瘤治疗,它红豆杉属的植物中有一种具有抗癌作用十分明显的结构相当复杂次生代谢产物。紫杉醇能通过与微管蛋白相结合,阻止微管解聚,阻断了细胞的正常分裂,使细胞分裂停止。因此,阻滞了细胞周期的正常运转,抑制癌细胞生长,从而起到抗癌作用。然而,紫杉醇治疗癌症的分子机理还不十分清楚。
硫氧还蛋白结合蛋白-2(Thioredoxin Binding Protein-2,TBP-2)是一种广泛存在于人体各组织中的分子量为46kD蛋白质。维生素D3能够诱导TBP-2在细胞中的表达量增加,所以,TBP-2又名叫维生素D3上调蛋白1(vitamin D3up regulated protein-1,VDUP-1), TBP-2与还原型硫氧还蛋白结合从而抑制其活性。当硫氧还蛋白半胱氨酸活性位点发生突变时硫氧还蛋白结合蛋白-2便不能与之相结合。因此,TBP-2是硫氧还蛋白的一个负性调节蛋白。通过在一些癌细胞中导入外源TBP-2表达质粒使其表达量增加,则可抑制细胞生长,促进细胞凋亡。TBP-2与糖代谢密切相关,葡萄糖可以诱导TBP-2的表达。
硫氧还蛋白(thioredoxin,Trx)是一种广泛分部在各种细胞和各种组织中的分子量为12kDa的多功能蛋白分子,硫氧还蛋白大约含有110个氨基酸这些氨基酸中有一个功能活性区域-Cys-Gly-Pro-Cys-,硫氧还蛋白同硫氧还蛋白还原酶(Thioredoxin reductase, TrxR)烟酰胺腺嘌呤二核苷酸磷酸(NADPH)三者一起组成硫氧还蛋白还原系统,一起调节某些转录因子的活性,调节细胞中的氧化还原反应的平衡。硫氧还蛋白还能与凋亡信号调节激酶1(apoptosis signal-regulating kinase1, ASK1)相结合,抑制细胞凋亡的途径。硫氧还蛋白还具有促进肿瘤血管再生和加速细胞生长等作用,与缺氧诱导因子及细胞周期蛋白关系密切。
本课题将使用将使用高浓度葡萄糖及紫杉醇刺激HepG2细胞,观察细胞活性变化,并且观察高糖是否促进紫杉醇诱导HepG2细胞凋亡；并研究紫杉醇作用后硫氧还蛋白,TBP-2,低氧诱导因子-1α,以及cyclinD1的表达；并使用SiRNA降低TBP-2的表达,检测在TBP-2低表达的情况下HepG2细胞对紫杉醇的敏感性。
研究结果：紫杉醇降低HepG2细胞活性,但高浓度葡萄糖并不促进紫杉醇的作用。紫杉醇抑制硫氧还蛋白及cyclinD1的表达。紫杉醇对TBP-2的表达无影响。TBP-2低表达的情况下对紫杉醇的抗癌作用具有抵抗作用性。
以上结果表明：紫杉醇通过抑制硫氧还蛋白,低氧诱导因子-1α以及cyclinD1的表达降低HepG2的活性,抑制其生长,促进凋亡。紫杉醇不具有诱导TBP-2的作用,但TBP-2低表达可以部分抵抗紫杉醇的作用。因此,硫氧还蛋白和TBP-2均可以作为紫杉醇的一个新的作用靶点。
Liver cancer is the malignant tumor in the liver, including primary liver cancer and metastatic liver cancer, liver cancer refers to people' daily said more than primary liver cancer. Primary liver cancer is one of the most common malignancies in the clinical, Primary liver cancer cells can be divided into the hepatocellular carcinoma, cholangiocarcinoma and mixed tumors. According to the latest statistics, around the world every year about six hundred thousand new cases of liver cancer patients, ranking fifth in the malignant tumor. There are many treatment methods on liver cancer,however,the effective mothod is few, chemotherapy is a widely used in treatment of liver cancer.
Paclitaxel is used as tumors in clinical, is a complex secondary metabolites in the genus Taxus, and is currently understood only one that can promote microtubules polymerization and stability of the polymerized microtubule drugs. Isotope tracing showed that Paclitaxel only binds to the polymerization of microtubules, inhibits its depolymerization, blocking the normal cell division, especially makes cell division stopped in mitosis, blocks the cell cycle, inhibits the cell growth, then plays role in anticancer. However, its molecular mechanisms on treating cancer are still unknown.
Thioredoxin binding protein-2is also called vitamin D3up regulated protein-1, or thioredoxin interacting protein, the molecular weight is46kDa, it was first isolated in HL-60cells, vitamin D3can induce the expression. TBP-2is the Trx endogenous inhibitor of Trx activities. TBP-2binds with the reduced state of Trx only, not bind with the oxidation state or activity site cysteine mutant, therefore, TBP-2is an inhibitor of Trx. TBP-2were transfected into some cells, the cell growth was inhibited, the apoptosis occurred. TBP-2is closely relate with glycometabolism. Glucose could induce the expression of TBP-2.
Thioredoxin is a molecular weight of12kDa protein, it has a highly conservative active site that is-Cys-Gly-Pro-Cys-, which has a variety of biological functions. Trx, NADPH and Thioredoxin reductase composed thioredoxin system, adjusting the intracellular oxidation-reduction equilibrium, regulating the activity of transcription factors. At the same time, Trx also combine with apoptosis signal-regulating kinase1,(ASK1), and also can promote the growth of cells and blood vessels regeneration. It is also associated with hypoxia-inducible factor-la (HIF-la) and cyclinD1.
This study will use high concentration of glucose and paclitaxel treated HepG2cells and detected cell viability, and observed whether high concentration of glucose enhanced the susceptibility of Paclitaxel in HepG2cells and promoted apoptosis by Paclitaxel. To examine the expressions of Thioredoxin, TBP-2HIFla and cyclinDl induced by Paclitaxel. By using SiRNA to decrease TBP-2expression, at lower expression of TBP-2, detected the HepG2cells susceptibility to Paclitaxel.
Results:Paclitaxel decreased cell viability in HepG2, but high concentration of glucose did not enhance the role of Paclitaxel. Paclitaxel inhibited the expressions of thioredoxin, cyclinDl and HIF-1α but not TBP-2. Lower expression of TBP-2had resistance to Paclitaxel.
Above results showed:Paclitaxel decreased cell viability, inhibited cell growth and induce apoptosis in HepG2by inhibiting the expressions of Thioredoxin, HIF-la and cyclinDl. Paclitaxel did not induce the expression of TBP-2, but the Lower expression of TBP-2had resistance to Paclitaxel.Thus, thioredoxin and TBP-2is a new target of Paclitaxel.
引文
[1]Flatter SJ.Bennelt GJ. Studies of peripheral sensory nerves in paclitaxel-induced painful peripheral neuropathy;Evidence for mitochondrial dysfunction [J].Pain,2006,122(3):245-252
[2]Regina A,Demeule M,Che C,et al Antitumour activity of ANG1005,a conjugate beween paclitaxel and the new brain delivery vector Angiopep-2 [j].Br J Pharmacol,2008,155 (2);185-190
[3]Nakayama S,Torikoshi Y,Takahashi T,et al.Prediction of paclitaxel sensitivity by CDK1 and CDK2 activity in human breast cancer cells [J].Breast Cancer Res,2009,11(1);R12
[4]Takahashi T,Yamasaki F,Sudo T,et al.CyclinA-associated kinase activity is needed for paclitaxel sensitivity [J].Mol Cancer Ther,2005,4(7):248-256
[5]Kgu-Ho-Han E, Gehrke L, Tahir SK, et ai. Moduiation of drug resistance by-tubuiin in paciitaxei-resistant human iung cancer ceii iines [J]. Eur J Cancer,2000, 36:1565-1571
[6]Ranganathan S, Dexter DW, Benetatos CA, et al. Cloning and seguencing of human-tubulin cDNA:induction of isotype in human prostate carcinoma cells by acute exposure to antimicrotubule agents [J]. Biochim Biophys Acta,1998,1395: 237-245.
[7]Ranganathan S, Benetatos CA,CoIarusso PJ, et al. Altered-tubulinisotype expression in paclitaxel-resistanthuman prostate carcinoma cells [J]. Br J Cancer, 1998,77(4):562-566.
[8]Kavallaris M, Burkhart CA, Horwitz SB.Antisense oligonucleotides to class-tubulin sensitize drug-resistant cells toTaxoI [J]. Br J Cancer,1999,80 (7):1020-1025.
[9]Kavallaris M, Kuo DYS, Burkhart CA, et al. Taxol-resistant epithelial ovarian tumors are associated with altered expression of specific-tubulin isotypes [J]. J Clin Invest,1997,100 (5):1282-1293.
[10]Banerjee A, Roach MC, Trcka P, et al. Increased microtubuie assembiy in bovine brain tubuiin iacking the type isotype of-tubuiin [J]. J Bioi Chem,1990, 265(3):1794-9 [11] Haber M, Burkhart CA, Regi DL, et ai.Altered expression of M 2, the ciass(32-tubuiin isotype in a murine J774.2 ceii iine with a high ievei of taxoi resistance [J]. J Bioi Chem,1995,270 (52):31269-31275.
[12]Jamieson L, Carpenter L, Biden TJ, etai. Protein kinese C activity is necessary for Bcr-Abi-mediated resistence to drug-induced apoptosis [J]. J Bioi Chem,1999,274 (7):3927-3930.
[13]Murray NR, Fieids AP. Atypicai protein kinase C iota protects human ieukemia ceiis against drug-induced apoptosis [J]. J Bioi Chem,1997,272 (44): 27521-27524.
[14]Cesaro P, Raiteri E, Demoz M, et ai.Experession of protein kinase C betai confers resistance to TNF aipha-and paciitaxei-induced apoptosis in HT-29 coion carcinoma ceiis [J]. Int J Cancer,2001,93 (2):179-184.
[15]Huang Y, Ibrado AM, Reed JC, et al.Co-expression of severai moiecuiar mechanisms of muitidrug resistance and their significance for paciitaxei cytotoxicity in human AML HL-60 ceiis [J]. Leukemia,1997,11:253-257.
[16]Chen K. S. and H. F. DeLuca, et al. Isolation and characterization of a novel cDNA from HL-60 cells treated with 1,25-dihydroxyvitamin D-3 [J]. Biochim Biophys Acta,1994,1219:26-32
[17]Bodnar J. S., et al. Positional cloning of the combined hyperlipidemia gene Hyplipl [J]. Nat Genet,2002,30:110-116
[18]Junn E., et al. Vitamin D-3 up-regulated protein 1 mediates oxidative stress via suppressing the thioredoxin function [J]. Journal of Immunology,2000,164: 6287-6295
[19]Champliaud M. F., et al. The expression of vitamin D-upregulated protein 1 in skin and its interaction with sciellin in cultured keratinocytes [J]. J Invest Dermatol, 2003,121:781-785
[20]Escrich E., et al. Identification of novel differentially expressed genes by the effect of a high-fat n-6 diet in experimental breast cancer [J]. Mol Carcinog,2004,40: 73-78
[21]Takahashi Y., et al. Localization of thioredoxin-interacting protein (TXNIP) mRNA in epithelium of human gastrointestinal tract [J]. J Histochem Cytochem,2003, 51:973-976
[22]Pahl H. L. and P. A. Baeuerle, et al. Oxygen and the control of gene expression [J]. Bioessays,1994,16:497-502
[23]Abate C., et al. Redox regulation of fos and jun DNA-binding activity in vitro [J]. Science,1990,249:1157-1161
[24]Toledano M. B. and W. J. Leonard, et al. Modulation of transcription factor NF-kappa B binding activity by oxidation-reduction in vitro [J]. Proc Natl Acad Sci U S A,1991,88:4328-4332
[25]Kagoshima H., et al. Functional dissection of the alpha and beta subunits of transcription factor PEBP2 and the redox susceptibility of its DNA binding activity [J]. J Biol Chem,1996,271:33074-33082
[26]Makino Y., et al. Thioredoxin:a redox-regulating cellular cofactor for glucocorticoid hormone action. Cross talk between endocrine control of stress response and cellular antioxidant defense system [J]. J Clin Invest,1996,98: 2469-2477
[27]Hayashi S., et al. Functional modulation of estrogen receptor by redox state with reference to thioredoxin as a mediator [J]. Nucleic Acids Res,1997,25: 4035-4040
[28]Kroemer G., et al. Mitochondrial control of apoptosis [J]. Immunol Today, 1997,18:44-51
[29]Ueda S., et al. Redox regulation of caspase-3(-like) protease activity: regulatory roles of thioredoxin and cytochrome c [J]. J Immunol,1998,161: 6689-6695
[30]Nishiyama A., et al. Identification of thioredoxin-binding protein-2/vitamin D(3) up-regulated protein 1 as a negative regulator of thioredoxin function and expression [J]. J Biol Chem,1999,274:21645-21650
[31]Yamanaka H., et al. A possible interaction of thioredoxin with VDUP1 in HeLa cells detected in a yeast two-hybrid system [J]. Biochemical and Biophysical Research Communications,2000,271:796-800
[32]Dutta K. K., et al. Two distinct mechanisms for loss of thioredoxin-binding protein-2 in oxidative stress-induced renal carcinogenesis [J]. Lab Invest,2005,85: 798-807
[33]HAN SH, Jeon JH, Ju HR et al VDUP1.upregulateed by TGF-betal and 1,25-dihydorxyvitamin D-3 inhibits tumor cell growth by blocking cell-cycle progressuon [J]. Oncogene,2003,22(26):4035-4046
[34]Nishinaka Y., et al. Loss of thioredoxin-binding protein-2/vitamin D3 up-regulated protein 1 in human T-cell leukemia virus type I-dependent T-cell transformation:implications for adult T-cell leukemia leukemogenesis [J]. Cancer Res, 2004,64:1287-1292
[35]G Chen, X., Tang, W., Liu, S., Yu, L., and Chen, Z. Thioredoxin-1 phosphorylated at T100 is needed for its anti-apoptotic activity in HepG2cancer cells [J]. Life Sci.2010,87:254-260.
[36]Oh, J.H., Chung, A.S., Steinbrenner, H., Sies, H., and Brenneisen, P.2004. Thioredoxin secreted upon ultraviolet A irradiation modulates activities of matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase-2 in human dermal fibroblasts [J]. Arch Biochem Biophys.2004.423:218-226.
[37]Lee, E., Jeong, J., Kim, S.E., Song, E.J., Kang, S.W., and Lee, K.J.. Multiple functions of Nm23-Hl are regulated by oxido-reduction system [J]. PLoS One.2009,4:e7949.
[38].Kase, S., Osaki, M., Honjo, S., Hashimoto, K., Adachi, H., Tsujitani, S., and Ito, H.. Expression of cyclooxygenase-1 and cyclooxygenase-2 in human esophageal mucosa, dysplasia and carcinoma [J]. Pathobiology.2004,71:84-92.
[39]Bi, N., Yang, M., Zhang, L., Chen, X., Ji, W., Ou, G., Lin, D., and Wang, L. Cyclooxygenase-2 genetic variants are associated with survival in unresectable locally advanced non-small cell lung cancer [J]. Clin Cancer Res.2010,16:2383-2390.
[40]Sudbo, J., Ristimaki, A., Sondresen, J.E., Kildal, W., Boysen, M., Koppang, H.S., Reith, A., Risberg, B., Nesland, J.M., and Bryne, M.. Cyclooxygenase-2 (COX-2) expression in high-risk premalignant oral lesions [J]. Oral Oncol,2003,39: 497-505.
[41]von Rahden, B.H., Stein, H.J., Puhringer, F., Koch, I., Langer, R., Piontek, G., Siewert, J.R., Hofler, H., and Sarbia, M. Coexpression of cyclooxygenases (COX-1, COX-2) and vascular endothelial growth factors (VEGF-A, VEGF-C) in esophageal adenocarcinoma [J]. Cancer Res,2005.65:5038-5044.
[42]Csiki, I., Yanagisawa, K., Haruki, N., Nadaf, S., Morrow, J.D., Johnson, D.H., and Carbone, D.P. Thioredoxin-1 modulates transcription of cyclooxygenase-2 via hypoxia-inducible factor-1 alpha in non-small cell lung cancer [J]. Cancer Res 2006,66:143-150.
[43]Welsh, S.J., Williams, R.R., Birmingham, A., Newman, D.J., Kirkpatrick,D.L., and Powis, G.. The thioredoxin redox inhibitors 1-methylpropyl 2-imidazolyl disulfide and pleurotin inhibit hypoxia-induced factor 1 alpha and vascular endothelial growth factor formation[J]. Mol Cancer Ther,2003 2:235-243.
[44]Milovanova, T.N., Bhopale, V.M., Sorokina, E.M., Moore, J.S., Hunt, T.K., Hauer-Jensen, M., Velazquez, O.C., and Thorn, S.R.. Lactate stimulates vasculogenic stem cells via the thioredoxin system and engages an autocrine activation loop involving hypoxia-inducible factor 1 [J]. Mol Cell Biol,2008,28:6248-6261.
[45]Kaga, S., Zhan, L., Matsumoto, M., and Maulik, N., Resveratrol enhances neovascularization in the infarcted rat myocardium through the induction of thioredoxin-1, heme oxygenase-1 and vascular endothelial growth factor [J]. Mol Cell Cardiol,2005,39:813-822.
[46]Persad S, Attwell S, Gray V, et al. Regulation of protein kinase B/ Akt-serine 473 phosphorylation by integrin·linked kinase:criticalroles for kinase activity and amino acids arginine 211 and serine 343[J]. J Biol Chem,2001,276(29): 27462-27469.
[47]Reif S, Lang A, Lindquist JN, et al. The role of focal adhesion ki-Base-phosphatidylinositol 3-kinase-Akt signaling in hepatic stellate cell proliferation and type I collagen expression [J].J Biol Chem,2003,278(10):8083-8090.
[48]Jetzt A, Howe JA, Horn MT, et al.Adenoviral-mediated expression of a kinase—dead mutant of Akt induces apoptosis selectively itumor cells and suppresses tumor growth in mice [J]. Cancer Res,2003,63(20):6697-6706.
[49]. West KA, Brognard J, Clark AS, et al. Rapid Akt activation by nicotine and a tobacco carcinogen modulates the phenotype of normal human airway epithelial cells [J]. J Clin Invest,2003, 111(1):81-90.
[50]Kim D, Kim S, Koh H, et al. Akt/PKB promotes cancer cell invasion via increased motility and metallopmteinase production [J]. FASEB J,2001,15(11): 1953-1962.
[51]Grille SJ, Bellacosa A, Upson J, et al. The protein kinase Akt induces epithelial mesenchymal transition and promotes enhanced motility and invasiveness of squamous cell carcinoma lines [J]. Cancer Res,2003,63(9):2172-2178.
[52]Lindberg LE, Hedjazifar S, Baeckstrom D. c-erbB2-induced disrudtion of matrix adhesion and morphogenesis reveals anovel role for protein kinaseB as anegative regulator of alPha(2)beta(1)integrin function [J]. Mol Biol cell,2002; 13: 2894-2908
[53]Hartmann C, Bartels G, Gehlhaar C, Holtkamp N, von Deimling A. PIK3CA mutations in glioblas.toma multiforme [J]. Acta Neuropathol(befl),2005; 109:639-642
[54]Sliva D, Rizzo MT, English D. Phosphatidylinositol-3-kinase and NF—KaPPaB regulate motility of invasive MDA-MB-231 human breast cancer cells bv the secretion of urokinase-type plasminogen activator [J]. Biof chem,2002,277: 3150-3157
[55]Hiendlmeyer E, Regus S, Wassermann s, Hlubek F, Haynl A, Dimnler A, Koch C, Knoll C, van Beest M, Reuning U, Brabletz T, Kirchner, Jung A. Beta-catenin up-regulates the expression of the Ilrokinase plasminogen activator in human colorectal tumors [J]. Cancer Res,2004,64:1209-1214.
[56]. Li P, Gao Y, Ji Z, Zhang X, Xu Q, Li G, Guo Z, Zheng B, Guo X. Role of urokinase plasminogen activator and its receptor in metastasis and invasion of neuroblastoma [J]. Pediatr surg,2004,39:1512-1519.
[57]Chan Cf, Yau To, Jin DY, Wong cM, Fan St, NgIO. Evaluation of nuclear factor-kappaB, urokinasetype plasminogen activator and HBx and their clinicopathological significance in hepatocellular carcinoma [J]. Clin Cancer Res, 2004,10:4140-4149.
[58]Agarwal A, Das K, Lerner N, Sathe S, Cicek M, Casey G, sizemore N. The Akt/I kappa B kinase pathway Promotes angiogenic/metastatic gene expression in colorectalcancer by activating nuclear factor-kappa B and beta-catenin [J]. Oncogene,2005; 24:1021-1031.
[59]Chandrasekar N, Mohanam S, Gujrati M, OliveroWC, Dinh DH, Rao JS. Downregulation of uPA inhibits migration and P13k/Akt signaling in glioblastoma cells [J]. Oncogene,2003,22:392-400.
[60]KobayashiH, Suzuki M, Kanayama N, Terao T. Genetic down regulation of phosphoinositide-3-kinase by bil(unin correlates with suppression of invasion and metastasis in human ovarian cancer HRA cells [J]. Biol Chem,2004,279:6371-6379.
[61]Wang M, Vogel I, Kalthoff H. Correlation beween metastati potential and variants from cOlOrectal tumor ceuline HT-29 [J]. World Gastroenterol,2003, 9:2627-2631.
[62]Sliva D. Sigmling Pathways responsible for cancer cell invasion as targets for cancer therapv [J]. Curr Cancer Drug Targets,2004,4:327-336.
[63]Kobayashi H, Suzuki M, Kanayama N, Terao T. Genetic down-regulation of phosphoinositide-3-kinase by bikunin correlates with suppression of invasion and metastasis in human ovarian cancer HRA cells [J]. Biol Chem,2004,279:6371-6379.
[64]Katso R, okkenhaug K, Ahmadi K, White S, Tjmms J, Waterfield MD. Cellular function of phosphoinositide-3-kinases:implications for development, homeostasis and cancer [J]. Annu Rev Cell Den Biol,2001,17:615-675.
[65]Namiecinska M, Marciniak K, Nowak IZ. VEGF as an angiogenic, neurotrophic and neuroprotective factor [J]. Postepy Hig Med Dosw,2005,59: 573-583
[66]Tan PH, Xue SA, Manunta M, Beutelspacher SC, Fazekasova H, Shamsul Alam AK, McClure Mo, George AJ. Effect of vectors on human endothelial cell signal transduction implications for cardiovascular gene therapy [J]. Arterioscler Thromb Vase Biol,2005,265(15):134-142
[67]. Adanls JM. Cory S. The Bcl-2 proteinfamily:arbiter of cell survival [J]. Science,1998,281(5381):1322—1326
[68]. Yi X Yin XM Dong Z Inhibition of Bid-induced ap(tptosis hy Bcl-2:tBid insertion, Bax transloeation and Bax / Bak oligemerization suppressed [J]. Biol Chem,2003,278(19):16992-16999.
[69]. Xin M G, Deng X M. Nicotine inactivation of the proapoptotic function of bax through phosphorylation[J]. J Biol Chem,2005,280(11):10781—10789.
[70]Song G, Ouyang G L, Bao S D. The activation of Akt/PKB signaling pathway and cell survival [J]. J Cell Mol Med,2005,9(1):59-71.
[71]Sells SF, Wood DP, Jr, et al, Smokeless tobacco, oxidative sterss, apoptosis, and antioxidants in androgen-dependent prostatecells [J]. Cell Growth Differ,1994, 5:457-466
[72]Culmsee C, zhu Y, Krieglstein J, et al. Evidence for the involvement of Par-4 in ischemeic neurou cell death [J]. J Cereb Blood Flow Metab,2001,21(4): 334-343
[73]Sangeeta KC, Sandip KM, Vivek MR, et al. Par-4 transcripcionally regulates Bcl-2 though a WT1-binding sitc on the bcl-2 promoter [J]. J Biol chem, 2003.278(22):19995-20005
[74]Knobbe CB, Kieslich AT, Reifenberger G. Genetic alteration and expression of the phosphoinositol-3-kinase/Akt pathway genes PIK3CA and PIKE in human glioblastomas [J]. Neuropathol Appl Neurobiol,2005,31(5):486-490.
[75]Vandermocre F, El Yazidi-Belkoural, Adriaenssens E, et al.The antiapoptotic effect of fibroblast growth factor-2 is mediated through nuclear factur-r,B activation induced via interaction between Akt and IkB kinase-β in breast cancer cells [J]. Oncngene,2005,24(35):5482-5491.
[76]Asnaghi L, Bmno P, Priulla M, et al. mTOR:a pmtein kinase switching between life and death [J]. Pllamacol Res,2004,50(6):545-549.
[77]郭琳,王强.PB1-Akt-mToR信号传导通路与恶性肿瘤浸润和转移的研究进展[J].现代肿瘤医学,2009,17(8)：1585-1588.
[78]Ouglas AL, Failla B, cindy JY, et al. Frequent mutation of the PIK3CA gene in ovarian and breast cancer [J]. Clin Cancer Res,2005, 11(8):2875-2878.
[79]MicⅢ P, Downward J. MechamismsiOf diseasea:P13K/Akt signaling in gastmintestinal cancers [J]. J Gastroenterol,2005,43(10):1133-1139.
[80]Sourbier C, Lindner V, Lang H, et al. The phosphoinositide-3-kinase/Akt pathway:a new target in human renal cell carcinoma therapy [J]. Cancer Res.2006, 66(10):5130-5142.
[81]Zhang R, Xu Y, Ekman N, et al. Etk/Bmx Transactivates Vascular endothelial Growth Factor 2 and Recruits Phosphatidylinositol-3-kinase to Mediate the Tumor Necrosis Factor-induced Angiogenic Pathway [J]. J Biol chen,2003,278: 51267-51276
[82]Gdich JR, De P, Dey N, et al. A vasular targeted pan phosphoinositide-3-kinase inhibitor prodmg SF1126 with afltilumor and tiallgiogenic activity [J]. Cancer Res,2008,68(1):206-215.
[83]Fukazawa H, Nouguchi K, Murakammi Y, et al. Mitogen activated protein/extracellular signal—regulated kinase kinase(MEK)inhibitors restore anoikis sensitivity in human breast criticer cell lines with a constitutively activated extracellular-regulated kinase(ERK)pathway [J]. Mol Cancer Ther,2002,1(5): 303-309.
[84]Cherian MG. Inhibition of extracellular signal regulated kinase(ERK)leads to apoptosis inducing factor(AIF)mediated apoptosis in epithelial breast caacer cells: the lack of effect of ERK in p53 mediated copper induced apoptosis [J]. J Cell Biochem,2005,95(6):1120-1134.
[85]Siddiqa A, Long LM, UL, et al. Expression of HER-2 in MCF-7 breast cancer cells modulates anti-apoptotic proteinsSurvivin and Bcl-2 via the extracellular signal-related kinase(ERK) and phosphoinositide 3-kinase (P13K) signalling pathways [J]. BMC Cancer,2008,8(8):129-138
[86]. Upadnyay AK, Ajak AK, Sing S, et al. Cell cycle regulatory protein 5 (Cdk5) is a novel downstream target of ERK in Carboplatin induced death of breast cancer cells [J]. Curt Cancer Drug Targets,2008,8(8):741-752.
[87]Yamamoto T, Ebisuya M, Ashida F, et al·Continuous ERK activation downregulates antiprolifemtive genes throughout G1 phase to allow cell cycle progression [J]. Curt Bid,2006,16(12):1171-1182.
[88]Chiu LC, Kong CK. The elllorophyllin—induced cell cycle arrest and apoptosis in human breast cancer MCF-7 cells is associated with ERK deactivation and Cyclin DI depletion [J]. Int J Mol Med,2005,16(4):735-740
[89]Eadahr SS, Dipp S, Baricos H. Bradykinin stimulates the ERK-EIk-1-Fos/AP-1 pathway in mesangial cells Am [J]. Physiol,1998,275(3 Pt 2): F343-352.
[90]Lester RD, Jom, Campana WM, et al. Erythropoietin promotes MCF-7 breast cancel cell migration by 811 ERK/mitogen-actlvated proteln kinase-dependent pathway and 1s priIrlanly responslbie for the Increase in migrration observed In hypoxia [J]. J Biol chem,2005,280(47):39273-39277.
[91]Hsiao PW, Chang CC, Liu HF, et al. Activation of p38 mitogen-activated protein kinase by celecoxib oppositely regulates aurvivin and gamma-H2AX in human eolorectal cancer cells [J].Toxicol Appl Pharmacol,2007,222(l):97-104
[92]Iyada K, Sasaki Y, et al. Involvement of the p38 mitogen-activated Drotein kinase cascade in hepatoceUular careinoma [J]. Cancer,2003,97(12):3017-3026.
[93]Paine E, Palmantier R, Akiyama SK, et al. Arachidonic acid activates miton activated prolein (MAP) kinase-activated protein kinase 2 and mediates adhesion of a human breast carcinma cell line to coullagen type IV through a p38 MAP kinase-dependent pathway [J]. J Biol Chem,2000,275(15):11284-11290.
[94]Kim MS, Lee EJ, Kim HR, et al. Bmkl/Erk5 is required for cell proliferation induced by epidermal growth factor [J].Nature,1998,395(6703):713-716
[95]Goncharova EA, Vorotnikov AV, Gracheva EO. Activation of p38 MAPkinase and caldesmon phosphorylation are essential for urokinase-induced human smooth muscle cell migration [J]. Bio Chem,2002,383(1):115
[96]Yu J, Bian D, Mahanivong C,et al. p38 Mitogen-activted protein kinase regulation of endothelial cell migration depends on urokinase plasminogen actvator expression [J]. Biol Chem,2000; 279, (48):50446
[97]Uu Si, Huang CC, Huang CJ, et al. Thimerosal induced apoptosis in human SCMI gastric cancer ceils:activation of p38 MAP kinase and caspase-3 pathways without involvement of [Ca2+]i elevation [J].Toxieol Sci,2007,100(1):109-117.
[98]Saeki K, Kobayshi N, Inazawa Y, et al. Oxidation triggered c-Jun N-terminal kinase (JNK)and p38 mitogenactivated protein(MAP)kinase pathways for apoptosis in human leukaemic cells stimulated by epigallocatechin-3-gallate(EGCG): a distinct pathway from those of chemically induced and receptor-mediated apaptosis [J]. Biochem J,2002,368:705-720.
[99]Abdel-latif MM, Athi-Morales V, et al. Deoxycholate induces COX-2 expression via Erkl/2, p38-MAPK and AP-1-dependent mechanisms in esophageal cancer cells[J]. BMC Cancer,2009,9:190
[100]Haddad J J. VX-745. Vertex pharmaceuticals [J]. Curt Opin Investig Drugs, 2001,2(8):1070-1076.
[101]Malemud C J. Inhibitora of stress-activated protein/mitogen-activel-ed protein kinase pathways [J].Curr Opin Pharmacol,2007,7(3):339-343.
[102]Lu Q, Qiu T Q, Yang H. Lignstilide inhibits vascular smooth musele cells proliferation [J]. Eur J Phannacol,2006,542(Ⅰ-3):136-140
[103]Wang J, Huang H, Liu P, et al. Inhibition of phesphorylation of p38 MAPK involved in the protection of nephropathy by emodin in diabetic rats [J]. Eur J Pharmacol,2006,553(1-3):297-303.
[104]Morel C, Ibarz G, Oiry C, et al. Cross-interactions of two p38 mitogen activated protein (MAP) kinase inhibitors and two choleeystokinin (CCK) receptor antagonists with the CCKI receptor and p38 MAP kinase [J]. J Biol Chem,2005, 280(22):21384-21393.
[105]Ozdemir C, Akdis C A. Discontinued drugs in 2006:pulmonaryllergy, dermelologieal, gastrointestinal and arthritis drugs [J]. Expertpin Investig Drugs,2007. 16(9):1327-1344.
[106]Angell RM, Bamborough P, Cleaaby A, et al. Biphenyl amide p38 kinase inhibitors 1:discovery and binding mode [J]. Bioorg Medhem Lett,2008,18(1): 318-323.
[2]Regina A,Demeule M,Che C,et al Antitumour activity of ANG1005,a conjugate beween paclitaxel and the new brain delivery vector Angiopep-2 [j].Br J Pharmacol,2008,155 (2);185-190
[3]Nakayama S,Torikoshi Y,Takahashi T,et al.Prediction of paclitaxel sensitivity by CDK1 and CDK2 activity in human breast cancer cells [J].Breast Cancer Res,2009,11(1);R12
[4]Takahashi T,Yamasaki F,Sudo T,et al.CyclinA-associated kinase activity is needed for paclitaxel sensitivity [J].Mol Cancer Ther,2005,4(7):248-256
[5]Kgu-Ho-Han E, Gehrke L, Tahir SK, et ai. Moduiation of drug resistance by-tubuiin in paciitaxei-resistant human iung cancer ceii iines [J]. Eur J Cancer,2000, 36:1565-1571
[6]Ranganathan S, Dexter DW, Benetatos CA, et al. Cloning and seguencing of human-tubulin cDNA:induction of isotype in human prostate carcinoma cells by acute exposure to antimicrotubule agents [J]. Biochim Biophys Acta,1998,1395: 237-245.
[7]Ranganathan S, Benetatos CA,CoIarusso PJ, et al. Altered-tubulinisotype expression in paclitaxel-resistanthuman prostate carcinoma cells [J]. Br J Cancer, 1998,77(4):562-566.
[8]Kavallaris M, Burkhart CA, Horwitz SB.Antisense oligonucleotides to class-tubulin sensitize drug-resistant cells toTaxoI [J]. Br J Cancer,1999,80 (7):1020-1025.
[9]Kavallaris M, Kuo DYS, Burkhart CA, et al. Taxol-resistant epithelial ovarian tumors are associated with altered expression of specific-tubulin isotypes [J]. J Clin Invest,1997,100 (5):1282-1293.
[10]Banerjee A, Roach MC, Trcka P, et al. Increased microtubuie assembiy in bovine brain tubuiin iacking the type isotype of-tubuiin [J]. J Bioi Chem,1990, 265(3):1794-9 [11] Haber M, Burkhart CA, Regi DL, et ai.Altered expression of M 2, the ciass(32-tubuiin isotype in a murine J774.2 ceii iine with a high ievei of taxoi resistance [J]. J Bioi Chem,1995,270 (52):31269-31275.
[12]Jamieson L, Carpenter L, Biden TJ, etai. Protein kinese C activity is necessary for Bcr-Abi-mediated resistence to drug-induced apoptosis [J]. J Bioi Chem,1999,274 (7):3927-3930.
[13]Murray NR, Fieids AP. Atypicai protein kinase C iota protects human ieukemia ceiis against drug-induced apoptosis [J]. J Bioi Chem,1997,272 (44): 27521-27524.
[14]Cesaro P, Raiteri E, Demoz M, et ai.Experession of protein kinase C betai confers resistance to TNF aipha-and paciitaxei-induced apoptosis in HT-29 coion carcinoma ceiis [J]. Int J Cancer,2001,93 (2):179-184.
[15]Huang Y, Ibrado AM, Reed JC, et al.Co-expression of severai moiecuiar mechanisms of muitidrug resistance and their significance for paciitaxei cytotoxicity in human AML HL-60 ceiis [J]. Leukemia,1997,11:253-257.
[16]Chen K. S. and H. F. DeLuca, et al. Isolation and characterization of a novel cDNA from HL-60 cells treated with 1,25-dihydroxyvitamin D-3 [J]. Biochim Biophys Acta,1994,1219:26-32
[17]Bodnar J. S., et al. Positional cloning of the combined hyperlipidemia gene Hyplipl [J]. Nat Genet,2002,30:110-116
[18]Junn E., et al. Vitamin D-3 up-regulated protein 1 mediates oxidative stress via suppressing the thioredoxin function [J]. Journal of Immunology,2000,164: 6287-6295
[19]Champliaud M. F., et al. The expression of vitamin D-upregulated protein 1 in skin and its interaction with sciellin in cultured keratinocytes [J]. J Invest Dermatol, 2003,121:781-785
[20]Escrich E., et al. Identification of novel differentially expressed genes by the effect of a high-fat n-6 diet in experimental breast cancer [J]. Mol Carcinog,2004,40: 73-78
[21]Takahashi Y., et al. Localization of thioredoxin-interacting protein (TXNIP) mRNA in epithelium of human gastrointestinal tract [J]. J Histochem Cytochem,2003, 51:973-976
[22]Pahl H. L. and P. A. Baeuerle, et al. Oxygen and the control of gene expression [J]. Bioessays,1994,16:497-502
[23]Abate C., et al. Redox regulation of fos and jun DNA-binding activity in vitro [J]. Science,1990,249:1157-1161
[24]Toledano M. B. and W. J. Leonard, et al. Modulation of transcription factor NF-kappa B binding activity by oxidation-reduction in vitro [J]. Proc Natl Acad Sci U S A,1991,88:4328-4332
[25]Kagoshima H., et al. Functional dissection of the alpha and beta subunits of transcription factor PEBP2 and the redox susceptibility of its DNA binding activity [J]. J Biol Chem,1996,271:33074-33082
[26]Makino Y., et al. Thioredoxin:a redox-regulating cellular cofactor for glucocorticoid hormone action. Cross talk between endocrine control of stress response and cellular antioxidant defense system [J]. J Clin Invest,1996,98: 2469-2477
[27]Hayashi S., et al. Functional modulation of estrogen receptor by redox state with reference to thioredoxin as a mediator [J]. Nucleic Acids Res,1997,25: 4035-4040
[28]Kroemer G., et al. Mitochondrial control of apoptosis [J]. Immunol Today, 1997,18:44-51
[29]Ueda S., et al. Redox regulation of caspase-3(-like) protease activity: regulatory roles of thioredoxin and cytochrome c [J]. J Immunol,1998,161: 6689-6695
[30]Nishiyama A., et al. Identification of thioredoxin-binding protein-2/vitamin D(3) up-regulated protein 1 as a negative regulator of thioredoxin function and expression [J]. J Biol Chem,1999,274:21645-21650
[31]Yamanaka H., et al. A possible interaction of thioredoxin with VDUP1 in HeLa cells detected in a yeast two-hybrid system [J]. Biochemical and Biophysical Research Communications,2000,271:796-800
[32]Dutta K. K., et al. Two distinct mechanisms for loss of thioredoxin-binding protein-2 in oxidative stress-induced renal carcinogenesis [J]. Lab Invest,2005,85: 798-807
[33]HAN SH, Jeon JH, Ju HR et al VDUP1.upregulateed by TGF-betal and 1,25-dihydorxyvitamin D-3 inhibits tumor cell growth by blocking cell-cycle progressuon [J]. Oncogene,2003,22(26):4035-4046
[34]Nishinaka Y., et al. Loss of thioredoxin-binding protein-2/vitamin D3 up-regulated protein 1 in human T-cell leukemia virus type I-dependent T-cell transformation:implications for adult T-cell leukemia leukemogenesis [J]. Cancer Res, 2004,64:1287-1292
[35]G Chen, X., Tang, W., Liu, S., Yu, L., and Chen, Z. Thioredoxin-1 phosphorylated at T100 is needed for its anti-apoptotic activity in HepG2cancer cells [J]. Life Sci.2010,87:254-260.
[36]Oh, J.H., Chung, A.S., Steinbrenner, H., Sies, H., and Brenneisen, P.2004. Thioredoxin secreted upon ultraviolet A irradiation modulates activities of matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase-2 in human dermal fibroblasts [J]. Arch Biochem Biophys.2004.423:218-226.
[37]Lee, E., Jeong, J., Kim, S.E., Song, E.J., Kang, S.W., and Lee, K.J.. Multiple functions of Nm23-Hl are regulated by oxido-reduction system [J]. PLoS One.2009,4:e7949.
[38].Kase, S., Osaki, M., Honjo, S., Hashimoto, K., Adachi, H., Tsujitani, S., and Ito, H.. Expression of cyclooxygenase-1 and cyclooxygenase-2 in human esophageal mucosa, dysplasia and carcinoma [J]. Pathobiology.2004,71:84-92.
[39]Bi, N., Yang, M., Zhang, L., Chen, X., Ji, W., Ou, G., Lin, D., and Wang, L. Cyclooxygenase-2 genetic variants are associated with survival in unresectable locally advanced non-small cell lung cancer [J]. Clin Cancer Res.2010,16:2383-2390.
[40]Sudbo, J., Ristimaki, A., Sondresen, J.E., Kildal, W., Boysen, M., Koppang, H.S., Reith, A., Risberg, B., Nesland, J.M., and Bryne, M.. Cyclooxygenase-2 (COX-2) expression in high-risk premalignant oral lesions [J]. Oral Oncol,2003,39: 497-505.
[41]von Rahden, B.H., Stein, H.J., Puhringer, F., Koch, I., Langer, R., Piontek, G., Siewert, J.R., Hofler, H., and Sarbia, M. Coexpression of cyclooxygenases (COX-1, COX-2) and vascular endothelial growth factors (VEGF-A, VEGF-C) in esophageal adenocarcinoma [J]. Cancer Res,2005.65:5038-5044.
[42]Csiki, I., Yanagisawa, K., Haruki, N., Nadaf, S., Morrow, J.D., Johnson, D.H., and Carbone, D.P. Thioredoxin-1 modulates transcription of cyclooxygenase-2 via hypoxia-inducible factor-1 alpha in non-small cell lung cancer [J]. Cancer Res 2006,66:143-150.
[43]Welsh, S.J., Williams, R.R., Birmingham, A., Newman, D.J., Kirkpatrick,D.L., and Powis, G.. The thioredoxin redox inhibitors 1-methylpropyl 2-imidazolyl disulfide and pleurotin inhibit hypoxia-induced factor 1 alpha and vascular endothelial growth factor formation[J]. Mol Cancer Ther,2003 2:235-243.
[44]Milovanova, T.N., Bhopale, V.M., Sorokina, E.M., Moore, J.S., Hunt, T.K., Hauer-Jensen, M., Velazquez, O.C., and Thorn, S.R.. Lactate stimulates vasculogenic stem cells via the thioredoxin system and engages an autocrine activation loop involving hypoxia-inducible factor 1 [J]. Mol Cell Biol,2008,28:6248-6261.
[45]Kaga, S., Zhan, L., Matsumoto, M., and Maulik, N., Resveratrol enhances neovascularization in the infarcted rat myocardium through the induction of thioredoxin-1, heme oxygenase-1 and vascular endothelial growth factor [J]. Mol Cell Cardiol,2005,39:813-822.
[46]Persad S, Attwell S, Gray V, et al. Regulation of protein kinase B/ Akt-serine 473 phosphorylation by integrin·linked kinase:criticalroles for kinase activity and amino acids arginine 211 and serine 343[J]. J Biol Chem,2001,276(29): 27462-27469.
[47]Reif S, Lang A, Lindquist JN, et al. The role of focal adhesion ki-Base-phosphatidylinositol 3-kinase-Akt signaling in hepatic stellate cell proliferation and type I collagen expression [J].J Biol Chem,2003,278(10):8083-8090.
[48]Jetzt A, Howe JA, Horn MT, et al.Adenoviral-mediated expression of a kinase—dead mutant of Akt induces apoptosis selectively itumor cells and suppresses tumor growth in mice [J]. Cancer Res,2003,63(20):6697-6706.
[49]. West KA, Brognard J, Clark AS, et al. Rapid Akt activation by nicotine and a tobacco carcinogen modulates the phenotype of normal human airway epithelial cells [J]. J Clin Invest,2003, 111(1):81-90.
[50]Kim D, Kim S, Koh H, et al. Akt/PKB promotes cancer cell invasion via increased motility and metallopmteinase production [J]. FASEB J,2001,15(11): 1953-1962.
[51]Grille SJ, Bellacosa A, Upson J, et al. The protein kinase Akt induces epithelial mesenchymal transition and promotes enhanced motility and invasiveness of squamous cell carcinoma lines [J]. Cancer Res,2003,63(9):2172-2178.
[52]Lindberg LE, Hedjazifar S, Baeckstrom D. c-erbB2-induced disrudtion of matrix adhesion and morphogenesis reveals anovel role for protein kinaseB as anegative regulator of alPha(2)beta(1)integrin function [J]. Mol Biol cell,2002; 13: 2894-2908
[53]Hartmann C, Bartels G, Gehlhaar C, Holtkamp N, von Deimling A. PIK3CA mutations in glioblas.toma multiforme [J]. Acta Neuropathol(befl),2005; 109:639-642
[54]Sliva D, Rizzo MT, English D. Phosphatidylinositol-3-kinase and NF—KaPPaB regulate motility of invasive MDA-MB-231 human breast cancer cells bv the secretion of urokinase-type plasminogen activator [J]. Biof chem,2002,277: 3150-3157
[55]Hiendlmeyer E, Regus S, Wassermann s, Hlubek F, Haynl A, Dimnler A, Koch C, Knoll C, van Beest M, Reuning U, Brabletz T, Kirchner, Jung A. Beta-catenin up-regulates the expression of the Ilrokinase plasminogen activator in human colorectal tumors [J]. Cancer Res,2004,64:1209-1214.
[56]. Li P, Gao Y, Ji Z, Zhang X, Xu Q, Li G, Guo Z, Zheng B, Guo X. Role of urokinase plasminogen activator and its receptor in metastasis and invasion of neuroblastoma [J]. Pediatr surg,2004,39:1512-1519.
[57]Chan Cf, Yau To, Jin DY, Wong cM, Fan St, NgIO. Evaluation of nuclear factor-kappaB, urokinasetype plasminogen activator and HBx and their clinicopathological significance in hepatocellular carcinoma [J]. Clin Cancer Res, 2004,10:4140-4149.
[58]Agarwal A, Das K, Lerner N, Sathe S, Cicek M, Casey G, sizemore N. The Akt/I kappa B kinase pathway Promotes angiogenic/metastatic gene expression in colorectalcancer by activating nuclear factor-kappa B and beta-catenin [J]. Oncogene,2005; 24:1021-1031.
[59]Chandrasekar N, Mohanam S, Gujrati M, OliveroWC, Dinh DH, Rao JS. Downregulation of uPA inhibits migration and P13k/Akt signaling in glioblastoma cells [J]. Oncogene,2003,22:392-400.
[60]KobayashiH, Suzuki M, Kanayama N, Terao T. Genetic down regulation of phosphoinositide-3-kinase by bil(unin correlates with suppression of invasion and metastasis in human ovarian cancer HRA cells [J]. Biol Chem,2004,279:6371-6379.
[61]Wang M, Vogel I, Kalthoff H. Correlation beween metastati potential and variants from cOlOrectal tumor ceuline HT-29 [J]. World Gastroenterol,2003, 9:2627-2631.
[62]Sliva D. Sigmling Pathways responsible for cancer cell invasion as targets for cancer therapv [J]. Curr Cancer Drug Targets,2004,4:327-336.
[63]Kobayashi H, Suzuki M, Kanayama N, Terao T. Genetic down-regulation of phosphoinositide-3-kinase by bikunin correlates with suppression of invasion and metastasis in human ovarian cancer HRA cells [J]. Biol Chem,2004,279:6371-6379.
[64]Katso R, okkenhaug K, Ahmadi K, White S, Tjmms J, Waterfield MD. Cellular function of phosphoinositide-3-kinases:implications for development, homeostasis and cancer [J]. Annu Rev Cell Den Biol,2001,17:615-675.
[65]Namiecinska M, Marciniak K, Nowak IZ. VEGF as an angiogenic, neurotrophic and neuroprotective factor [J]. Postepy Hig Med Dosw,2005,59: 573-583
[66]Tan PH, Xue SA, Manunta M, Beutelspacher SC, Fazekasova H, Shamsul Alam AK, McClure Mo, George AJ. Effect of vectors on human endothelial cell signal transduction implications for cardiovascular gene therapy [J]. Arterioscler Thromb Vase Biol,2005,265(15):134-142
[67]. Adanls JM. Cory S. The Bcl-2 proteinfamily:arbiter of cell survival [J]. Science,1998,281(5381):1322—1326
[68]. Yi X Yin XM Dong Z Inhibition of Bid-induced ap(tptosis hy Bcl-2:tBid insertion, Bax transloeation and Bax / Bak oligemerization suppressed [J]. Biol Chem,2003,278(19):16992-16999.
[69]. Xin M G, Deng X M. Nicotine inactivation of the proapoptotic function of bax through phosphorylation[J]. J Biol Chem,2005,280(11):10781—10789.
[70]Song G, Ouyang G L, Bao S D. The activation of Akt/PKB signaling pathway and cell survival [J]. J Cell Mol Med,2005,9(1):59-71.
[71]Sells SF, Wood DP, Jr, et al, Smokeless tobacco, oxidative sterss, apoptosis, and antioxidants in androgen-dependent prostatecells [J]. Cell Growth Differ,1994, 5:457-466
[72]Culmsee C, zhu Y, Krieglstein J, et al. Evidence for the involvement of Par-4 in ischemeic neurou cell death [J]. J Cereb Blood Flow Metab,2001,21(4): 334-343
[73]Sangeeta KC, Sandip KM, Vivek MR, et al. Par-4 transcripcionally regulates Bcl-2 though a WT1-binding sitc on the bcl-2 promoter [J]. J Biol chem, 2003.278(22):19995-20005
[74]Knobbe CB, Kieslich AT, Reifenberger G. Genetic alteration and expression of the phosphoinositol-3-kinase/Akt pathway genes PIK3CA and PIKE in human glioblastomas [J]. Neuropathol Appl Neurobiol,2005,31(5):486-490.
[75]Vandermocre F, El Yazidi-Belkoural, Adriaenssens E, et al.The antiapoptotic effect of fibroblast growth factor-2 is mediated through nuclear factur-r,B activation induced via interaction between Akt and IkB kinase-β in breast cancer cells [J]. Oncngene,2005,24(35):5482-5491.
[76]Asnaghi L, Bmno P, Priulla M, et al. mTOR:a pmtein kinase switching between life and death [J]. Pllamacol Res,2004,50(6):545-549.
[77]郭琳,王强.PB1-Akt-mToR信号传导通路与恶性肿瘤浸润和转移的研究进展[J].现代肿瘤医学,2009,17(8)：1585-1588.
[78]Ouglas AL, Failla B, cindy JY, et al. Frequent mutation of the PIK3CA gene in ovarian and breast cancer [J]. Clin Cancer Res,2005, 11(8):2875-2878.
[79]MicⅢ P, Downward J. MechamismsiOf diseasea:P13K/Akt signaling in gastmintestinal cancers [J]. J Gastroenterol,2005,43(10):1133-1139.
[80]Sourbier C, Lindner V, Lang H, et al. The phosphoinositide-3-kinase/Akt pathway:a new target in human renal cell carcinoma therapy [J]. Cancer Res.2006, 66(10):5130-5142.
[81]Zhang R, Xu Y, Ekman N, et al. Etk/Bmx Transactivates Vascular endothelial Growth Factor 2 and Recruits Phosphatidylinositol-3-kinase to Mediate the Tumor Necrosis Factor-induced Angiogenic Pathway [J]. J Biol chen,2003,278: 51267-51276
[82]Gdich JR, De P, Dey N, et al. A vasular targeted pan phosphoinositide-3-kinase inhibitor prodmg SF1126 with afltilumor and tiallgiogenic activity [J]. Cancer Res,2008,68(1):206-215.
[83]Fukazawa H, Nouguchi K, Murakammi Y, et al. Mitogen activated protein/extracellular signal—regulated kinase kinase(MEK)inhibitors restore anoikis sensitivity in human breast criticer cell lines with a constitutively activated extracellular-regulated kinase(ERK)pathway [J]. Mol Cancer Ther,2002,1(5): 303-309.
[84]Cherian MG. Inhibition of extracellular signal regulated kinase(ERK)leads to apoptosis inducing factor(AIF)mediated apoptosis in epithelial breast caacer cells: the lack of effect of ERK in p53 mediated copper induced apoptosis [J]. J Cell Biochem,2005,95(6):1120-1134.
[85]Siddiqa A, Long LM, UL, et al. Expression of HER-2 in MCF-7 breast cancer cells modulates anti-apoptotic proteinsSurvivin and Bcl-2 via the extracellular signal-related kinase(ERK) and phosphoinositide 3-kinase (P13K) signalling pathways [J]. BMC Cancer,2008,8(8):129-138
[86]. Upadnyay AK, Ajak AK, Sing S, et al. Cell cycle regulatory protein 5 (Cdk5) is a novel downstream target of ERK in Carboplatin induced death of breast cancer cells [J]. Curt Cancer Drug Targets,2008,8(8):741-752.
[87]Yamamoto T, Ebisuya M, Ashida F, et al·Continuous ERK activation downregulates antiprolifemtive genes throughout G1 phase to allow cell cycle progression [J]. Curt Bid,2006,16(12):1171-1182.
[88]Chiu LC, Kong CK. The elllorophyllin—induced cell cycle arrest and apoptosis in human breast cancer MCF-7 cells is associated with ERK deactivation and Cyclin DI depletion [J]. Int J Mol Med,2005,16(4):735-740
[89]Eadahr SS, Dipp S, Baricos H. Bradykinin stimulates the ERK-EIk-1-Fos/AP-1 pathway in mesangial cells Am [J]. Physiol,1998,275(3 Pt 2): F343-352.
[90]Lester RD, Jom, Campana WM, et al. Erythropoietin promotes MCF-7 breast cancel cell migration by 811 ERK/mitogen-actlvated proteln kinase-dependent pathway and 1s priIrlanly responslbie for the Increase in migrration observed In hypoxia [J]. J Biol chem,2005,280(47):39273-39277.
[91]Hsiao PW, Chang CC, Liu HF, et al. Activation of p38 mitogen-activated protein kinase by celecoxib oppositely regulates aurvivin and gamma-H2AX in human eolorectal cancer cells [J].Toxicol Appl Pharmacol,2007,222(l):97-104
[92]Iyada K, Sasaki Y, et al. Involvement of the p38 mitogen-activated Drotein kinase cascade in hepatoceUular careinoma [J]. Cancer,2003,97(12):3017-3026.
[93]Paine E, Palmantier R, Akiyama SK, et al. Arachidonic acid activates miton activated prolein (MAP) kinase-activated protein kinase 2 and mediates adhesion of a human breast carcinma cell line to coullagen type IV through a p38 MAP kinase-dependent pathway [J]. J Biol Chem,2000,275(15):11284-11290.
[94]Kim MS, Lee EJ, Kim HR, et al. Bmkl/Erk5 is required for cell proliferation induced by epidermal growth factor [J].Nature,1998,395(6703):713-716
[95]Goncharova EA, Vorotnikov AV, Gracheva EO. Activation of p38 MAPkinase and caldesmon phosphorylation are essential for urokinase-induced human smooth muscle cell migration [J]. Bio Chem,2002,383(1):115
[96]Yu J, Bian D, Mahanivong C,et al. p38 Mitogen-activted protein kinase regulation of endothelial cell migration depends on urokinase plasminogen actvator expression [J]. Biol Chem,2000; 279, (48):50446
[97]Uu Si, Huang CC, Huang CJ, et al. Thimerosal induced apoptosis in human SCMI gastric cancer ceils:activation of p38 MAP kinase and caspase-3 pathways without involvement of [Ca2+]i elevation [J].Toxieol Sci,2007,100(1):109-117.
[98]Saeki K, Kobayshi N, Inazawa Y, et al. Oxidation triggered c-Jun N-terminal kinase (JNK)and p38 mitogenactivated protein(MAP)kinase pathways for apoptosis in human leukaemic cells stimulated by epigallocatechin-3-gallate(EGCG): a distinct pathway from those of chemically induced and receptor-mediated apaptosis [J]. Biochem J,2002,368:705-720.
[99]Abdel-latif MM, Athi-Morales V, et al. Deoxycholate induces COX-2 expression via Erkl/2, p38-MAPK and AP-1-dependent mechanisms in esophageal cancer cells[J]. BMC Cancer,2009,9:190
[100]Haddad J J. VX-745. Vertex pharmaceuticals [J]. Curt Opin Investig Drugs, 2001,2(8):1070-1076.
[101]Malemud C J. Inhibitora of stress-activated protein/mitogen-activel-ed protein kinase pathways [J].Curr Opin Pharmacol,2007,7(3):339-343.
[102]Lu Q, Qiu T Q, Yang H. Lignstilide inhibits vascular smooth musele cells proliferation [J]. Eur J Phannacol,2006,542(Ⅰ-3):136-140
[103]Wang J, Huang H, Liu P, et al. Inhibition of phesphorylation of p38 MAPK involved in the protection of nephropathy by emodin in diabetic rats [J]. Eur J Pharmacol,2006,553(1-3):297-303.
[104]Morel C, Ibarz G, Oiry C, et al. Cross-interactions of two p38 mitogen activated protein (MAP) kinase inhibitors and two choleeystokinin (CCK) receptor antagonists with the CCKI receptor and p38 MAP kinase [J]. J Biol Chem,2005, 280(22):21384-21393.
[105]Ozdemir C, Akdis C A. Discontinued drugs in 2006:pulmonaryllergy, dermelologieal, gastrointestinal and arthritis drugs [J]. Expertpin Investig Drugs,2007. 16(9):1327-1344.
[106]Angell RM, Bamborough P, Cleaaby A, et al. Biphenyl amide p38 kinase inhibitors 1:discovery and binding mode [J]. Bioorg Medhem Lett,2008,18(1): 318-323.