Runx3，Smad4及Ki-67蛋白在皮肤恶性黑色素瘤中的表达和意义
摘要
目的
皮肤恶性黑色素瘤(cutaneous malignant melanoma, CMM)是一种恶性程度极高的皮肤肿瘤,细胞无限增殖与凋亡抑制是其主要发病机制。目前对CMM尚无完全有效的预防与治疗措施。
国内外对于CMM发生机制的研究已经取得了较大的进步。研究表明,CMM是正常细胞染色体多重损伤的复杂过程,它的发生、发展以及转归的研究涉及了癌基因的激活、抑癌基因的失活、凋亡基因紊乱、DNA转录表达失控等。人类runt相关转录因子3(runt-related transcription factor 3 gene, Runx3)作为一种抑癌基因,其蛋白可指导转化生长因子-β(transforming growth factor-β, TGF-β)信号转导过程中激活的Smad复合物从细胞质内转入细胞核内的特定靶位点,加强Smad复合物与靶位点的结合强度并激活靶基因,从而对细胞的分化、细胞周期调控、凋亡和恶性转化起作用。Ki-67是与增殖细胞相关的核抗原。本实验通过检测Runx3、Smad4及Ki-67蛋白在CMM中的表达水平并分析它们之间的相互关系,从细胞增殖和凋亡方面探讨Runx3、Smad4及Ki-67对CMM发生发展所起的作用。
材料与方法
本研究选用郑州大学第一附属医院病理科2006年7月～2008年12月切除且病历资料完整、病理诊断明确的CMM38例及皮肤交界痣20例；所有患者均无合并皮肤病、结缔组织病、免疫系统疾病及其他重要脏器疾病；患者术前均未进行放疗、化疗、免疫、冷冻、激光及其他治疗。20例正常皮肤组织(对照组)为郑州大学第一附属医院整形外科非癌及未合并皮肤病的患者行全厚皮片移植时修剪所剩全厚皮片与美容手术者切除的多余全厚皮肤。运用免疫组织化学SP法,探讨在CMM中Runx3, Smad4和与细胞增殖相关的Ki-67之间的关系。统计方法采用SPSS13.0软件对数据进行处理,运用卡方检验和相关性分析,检验标准α=0.05,X2分割检验标准α’=0.0167。
结果
1.Runx3蛋白主要表达于黑色素细胞的细胞核,在CMM中低表达。在CMM中其阳性表达率为23.68%(9／38),与皮肤交界痣80%(16／20)、正常皮肤90%(18／20)相比差异具有统计学意义(P<0.0167),皮肤交界痣与正常皮肤相比差异无统计学意义(P>0.0167)
2.Smad4蛋白主要表达于黑色素细胞的细胞质,在CMM中低表达。在CMM中其阳性表达率为34.21%(13／38),与皮肤交界痣85%(17／20)、正常皮肤95%(19／20)相比差异具有统计学意义(P<0.0167),皮肤交界痣与正常皮肤相比差异无统计学意义(P>0.0167)。
3.Ki-67蛋白主要表达于黑色素细胞即增殖细胞的细胞核。在CMM中其阳性表达率为78.95%(30／38),与皮肤交界痣30.00%(6／20)、正常皮肤25%(5／20)相比差异具有统计学意义(P<0.0167),皮肤交界痣与正常皮肤相比差异无统计学意义(P>0.0167)。
4.相关性分析显示,Runx3、Smad4蛋白表达与细胞增殖核抗原Ki-67蛋白表达呈负相关(P<0.05),Runx3与Smad4蛋白表达呈正相关(r=0.455)。
结论
1.Runx3与Smad4蛋白在CMM黑色素细胞中的表达明显较皮肤交界痣及正常皮肤低,它们的低表达可能通过某种细胞信号转导途径促进黑色素细胞的增殖,抑制黑色素细胞的凋亡,从而促进CMM的形成与发展。提示它们在CMM中可能起抑癌作用。
2.Smad4参与的TGF-β信号转导途径最终作用是促进细胞凋亡,抑制细胞增殖。Runx3是其上游的作用分子,在CMM中其蛋白低表达可阻碍该信号途径的顺利进行,从而促进CMM的发展。
3.Ki-67是一种细胞增殖核抗原,在CMM中可反映黑色素细胞的增殖活性,为CMM的发展程度和预后提供可靠的分子依据。
Objective
Cutaneous malignant melanoma (CMM) is a highly malignant tumor of skin. Its main pathogenesis are cell proliferation and infinite apoptosis. There is no effective prevention and therapy measurement for CMM at present.
The researches on the mechanism of CMM showed that CMM is a complex process of multiple injuries in the normal chromosome'. The development, growth and turnover refer to the oncogene activation, tumor suppressor gene inactivation, apoptosis gene disorders, uncontrolled DNA transcription and so on. As a suppressor gene of tumor, its protein can direct the Smad complex from the cytoplasm translate into the nucleus in the TGF-P signal transduction to activate the specific target sites, enhance Smad complex together with the target site of adhesion strength and cell differentiation, cell cycle regulation, apoptosis and malignant transformation of work. Ki-67 is a nuclear antigen that associated with the proliferation. In this research immunohistochemical technique is used to study the expression of Runx3, Smad4, and their relations with the Ki-67 which reflect the proliferation activity, in order to explore Runx3, Smad4 and Ki-67'effection during the generation and development of CMM.
Materials and methods
38 cases CMM and 20 cases normal skin with complete resection and clear pathological diagnosis were selected which came from the department of pathologyinthe first affiliated hospital of Zhengzhou University. All patients had no merger skin, connective tissue disease, immune disorders and other important organs diseases; Patients were not carried out prior to radiotherapy, chemotherapy, immune, freezing, laser and other treatment.20 cases of normal skin which came from full thick skin grafts were chosen as control groups. The SP immunohistochemical technique was used to detect the expression of Runx3, Smad4 with cell proliferation in the development of CMM. The statistics data were processed by SPSS 13.0, using Chi-square test and Dependability analysis. The test standard is 0.05 and the standard of x2 division is 0.0167.
Results
1. Runx3 expressed mainly on the nucleus of melanoma cells. There was a statical significance between the CMM and mole skin junction or normal skin(P<0.0167) There is no statistical significance between mole skin junction and normal skin (P> 0.0167)
2. Smad4 expressed mainly on the cytoplasm of melanoma cells. There was a statical significance between the CMM and mole skin junction or normal skin (P<0.0167) There was no statistical significance between mole skin junction and normal skin (P >0.0167)
3. Ki-67 expressed mainly on the nucleus of melanoma cells. There was a statical significance between the CMM and mole skin junction or normal skin(P<0.0167) There was no statistical significance between mole skin junction and normal skin (P >0.0167)
4. The dependability analysis shoewed that there was a significant negative correlation between Runx3 and Smad4 and Ki-67(P<0.05). There was a significant positive correlation between Runx3 and Smad4(r=0.455).
Conclusions
1. It is speculated that Runx3 and Smad4 might play a tumor suppressor role in CMM for their low expressions. The low expression may promote the formation and development of CMM by promoting cell proliferation and inhibiting cell apoptosis through the cell signaling pathways.
2. The final effect of the TGF-βsignaling system is to promote cell apoptosis and inhibit cell proliferation. As an upstream element, Runx3 can inhibit the development of CMM by guiding the signaling pathway successfully.
3. Ki-67 can reflect the proliferation of melanocytes and provide the molecular basis of the degree for the prognosis and development of CMM.
皮肤恶性黑色素瘤(cutaneous malignant melanoma, CMM)是一种恶性程度极高的皮肤肿瘤,细胞无限增殖与凋亡抑制是其主要发病机制。目前对CMM尚无完全有效的预防与治疗措施。
国内外对于CMM发生机制的研究已经取得了较大的进步。研究表明,CMM是正常细胞染色体多重损伤的复杂过程,它的发生、发展以及转归的研究涉及了癌基因的激活、抑癌基因的失活、凋亡基因紊乱、DNA转录表达失控等。人类runt相关转录因子3(runt-related transcription factor 3 gene, Runx3)作为一种抑癌基因,其蛋白可指导转化生长因子-β(transforming growth factor-β, TGF-β)信号转导过程中激活的Smad复合物从细胞质内转入细胞核内的特定靶位点,加强Smad复合物与靶位点的结合强度并激活靶基因,从而对细胞的分化、细胞周期调控、凋亡和恶性转化起作用。Ki-67是与增殖细胞相关的核抗原。本实验通过检测Runx3、Smad4及Ki-67蛋白在CMM中的表达水平并分析它们之间的相互关系,从细胞增殖和凋亡方面探讨Runx3、Smad4及Ki-67对CMM发生发展所起的作用。
材料与方法
本研究选用郑州大学第一附属医院病理科2006年7月～2008年12月切除且病历资料完整、病理诊断明确的CMM38例及皮肤交界痣20例；所有患者均无合并皮肤病、结缔组织病、免疫系统疾病及其他重要脏器疾病；患者术前均未进行放疗、化疗、免疫、冷冻、激光及其他治疗。20例正常皮肤组织(对照组)为郑州大学第一附属医院整形外科非癌及未合并皮肤病的患者行全厚皮片移植时修剪所剩全厚皮片与美容手术者切除的多余全厚皮肤。运用免疫组织化学SP法,探讨在CMM中Runx3, Smad4和与细胞增殖相关的Ki-67之间的关系。统计方法采用SPSS13.0软件对数据进行处理,运用卡方检验和相关性分析,检验标准α=0.05,X2分割检验标准α’=0.0167。
结果
1.Runx3蛋白主要表达于黑色素细胞的细胞核,在CMM中低表达。在CMM中其阳性表达率为23.68%(9／38),与皮肤交界痣80%(16／20)、正常皮肤90%(18／20)相比差异具有统计学意义(P<0.0167),皮肤交界痣与正常皮肤相比差异无统计学意义(P>0.0167)
2.Smad4蛋白主要表达于黑色素细胞的细胞质,在CMM中低表达。在CMM中其阳性表达率为34.21%(13／38),与皮肤交界痣85%(17／20)、正常皮肤95%(19／20)相比差异具有统计学意义(P<0.0167),皮肤交界痣与正常皮肤相比差异无统计学意义(P>0.0167)。
3.Ki-67蛋白主要表达于黑色素细胞即增殖细胞的细胞核。在CMM中其阳性表达率为78.95%(30／38),与皮肤交界痣30.00%(6／20)、正常皮肤25%(5／20)相比差异具有统计学意义(P<0.0167),皮肤交界痣与正常皮肤相比差异无统计学意义(P>0.0167)。
4.相关性分析显示,Runx3、Smad4蛋白表达与细胞增殖核抗原Ki-67蛋白表达呈负相关(P<0.05),Runx3与Smad4蛋白表达呈正相关(r=0.455)。
结论
1.Runx3与Smad4蛋白在CMM黑色素细胞中的表达明显较皮肤交界痣及正常皮肤低,它们的低表达可能通过某种细胞信号转导途径促进黑色素细胞的增殖,抑制黑色素细胞的凋亡,从而促进CMM的形成与发展。提示它们在CMM中可能起抑癌作用。
2.Smad4参与的TGF-β信号转导途径最终作用是促进细胞凋亡,抑制细胞增殖。Runx3是其上游的作用分子,在CMM中其蛋白低表达可阻碍该信号途径的顺利进行,从而促进CMM的发展。
3.Ki-67是一种细胞增殖核抗原,在CMM中可反映黑色素细胞的增殖活性,为CMM的发展程度和预后提供可靠的分子依据。
Objective
Cutaneous malignant melanoma (CMM) is a highly malignant tumor of skin. Its main pathogenesis are cell proliferation and infinite apoptosis. There is no effective prevention and therapy measurement for CMM at present.
The researches on the mechanism of CMM showed that CMM is a complex process of multiple injuries in the normal chromosome'. The development, growth and turnover refer to the oncogene activation, tumor suppressor gene inactivation, apoptosis gene disorders, uncontrolled DNA transcription and so on. As a suppressor gene of tumor, its protein can direct the Smad complex from the cytoplasm translate into the nucleus in the TGF-P signal transduction to activate the specific target sites, enhance Smad complex together with the target site of adhesion strength and cell differentiation, cell cycle regulation, apoptosis and malignant transformation of work. Ki-67 is a nuclear antigen that associated with the proliferation. In this research immunohistochemical technique is used to study the expression of Runx3, Smad4, and their relations with the Ki-67 which reflect the proliferation activity, in order to explore Runx3, Smad4 and Ki-67'effection during the generation and development of CMM.
Materials and methods
38 cases CMM and 20 cases normal skin with complete resection and clear pathological diagnosis were selected which came from the department of pathologyinthe first affiliated hospital of Zhengzhou University. All patients had no merger skin, connective tissue disease, immune disorders and other important organs diseases; Patients were not carried out prior to radiotherapy, chemotherapy, immune, freezing, laser and other treatment.20 cases of normal skin which came from full thick skin grafts were chosen as control groups. The SP immunohistochemical technique was used to detect the expression of Runx3, Smad4 with cell proliferation in the development of CMM. The statistics data were processed by SPSS 13.0, using Chi-square test and Dependability analysis. The test standard is 0.05 and the standard of x2 division is 0.0167.
Results
1. Runx3 expressed mainly on the nucleus of melanoma cells. There was a statical significance between the CMM and mole skin junction or normal skin(P<0.0167) There is no statistical significance between mole skin junction and normal skin (P> 0.0167)
2. Smad4 expressed mainly on the cytoplasm of melanoma cells. There was a statical significance between the CMM and mole skin junction or normal skin (P<0.0167) There was no statistical significance between mole skin junction and normal skin (P >0.0167)
3. Ki-67 expressed mainly on the nucleus of melanoma cells. There was a statical significance between the CMM and mole skin junction or normal skin(P<0.0167) There was no statistical significance between mole skin junction and normal skin (P >0.0167)
4. The dependability analysis shoewed that there was a significant negative correlation between Runx3 and Smad4 and Ki-67(P<0.05). There was a significant positive correlation between Runx3 and Smad4(r=0.455).
Conclusions
1. It is speculated that Runx3 and Smad4 might play a tumor suppressor role in CMM for their low expressions. The low expression may promote the formation and development of CMM by promoting cell proliferation and inhibiting cell apoptosis through the cell signaling pathways.
2. The final effect of the TGF-βsignaling system is to promote cell apoptosis and inhibit cell proliferation. As an upstream element, Runx3 can inhibit the development of CMM by guiding the signaling pathway successfully.
3. Ki-67 can reflect the proliferation of melanocytes and provide the molecular basis of the degree for the prognosis and development of CMM.
引文
[1]Slominski A, Wortsman J, Carlson AJ, et al. Malignant melanoma [J]. Arch PatholLabMed, 2001,125(10):1295-306
[2]RiesLAG,EisnerMP,Kosary,etaleditors.SEERCancerStatisticsREVIEW,1973-2001.Bethesda, MD.NationalCancerInstitute,2004.Availablefrom:http://seer,cancer.gov/csr/1975-2001.Accesses May,2004
[3]Kang GH, Lee S, Lee HJ, Hwang KS. Aberrant CpG island hypermethyaltion ofmultiple genes in prostatic cacenr and prostatic intraepithelial neoplasia [J]. J Pathol 2004;202:233-240
[4]Kjm TY, Lee HJ, Hwang KS, Lee M.Kim JW, Bang YJ, Kang GH. Methylation of Runx3 in various types of human cancers premalignant, stages of gastric carcinoma [J] Lab Invest 2004;84:479-484
[5]Wada M, Yazumi S, Takaishi S, Hasegawa K, Sawada M, Tanaka H, Ida H, SakakuraC, Ito Y, Chiba T. Frequent loss of Runx3 gene expression in human bile duct andPancreatic cancer cell lincs [J]. Oncogene 2004;23:2401-2407
[6]Li QL, Kim WJ, Choi JK, Lee YH, Kim HM, Li LS, Kim H Chang J, ItoY, Youl LeeK.Bae SC. TranscriPtional silencing of the Runx3 gene by CpG hypermethylation isAssociated with lung cancer [J]. Biochem Biophys Res Commun 2004;314:223-228
[7]Schulze-Bergkamen H, Krammer PH. Apoptosis in cancer implication for therapy [J]. Semin Oncol 2004;31:90-119
[8]Li QL, Ito K, Sakakura C, et al. Causal relationship between the loss of Runx3 expression and gastric cancer [J].Cell,2002,109(1):113-124
[9]Baylin SB, Belinsky SA, Herman JG. Aberrant methylation of gene promoters inCancer concepts, misconcepts and, Promise [J].JNatl Cancer, Inst,2000,92:1406-1461
[10]Ito Y, Miyazono K. Runx3 transcription factors as key targets of TGF-beta superfamily signaling [J]. Currop in GenetDev,2003,13:34-47
[11]Zaidi SK, Sullivan AJ, van Wijnen AJ, et al. Integration of Runx and smad regulatorySignals at tannscriptionally active subnuclear sites [J] ProcNatlAcad SciUSA,2002,99(12):8048-8053
[12]Kato N, Tamura G, Fukase M, et al.HyPermethylation of the Runx3 gene Promoterin testicular yolk sactumor of infants [J], Am J Pathol,2003,163(2):387-398
[13]Schwarte-Waldhoff I, Schmiegel W. Smad4 transcriptional path-ways and angiogenesis[J]. Int J Gastrointest Cancer,2002,31(1):47-59
[14]Schwarte-Waldhoff 1, Klein S, Blass-Kampmann S, et al. DPC4/Smad4 mediated tumor supp-ression of colon carcinorma cells is associated with reduced urokinses expression[J]. Oncogene,1999,18(26):3152-3158
[15]Asakura T, Takano Y, Iki M, et al. Prognostic value of Ki-67 for recurrence andprogression of superficial bladder cancer[J]. J Urol,1997,158(2):385-388
[16]孙妍,曹玉文,陆天才,等.宫颈上皮癌变过程中细胞分化的研究[J].癌症,2005,24(10)：1184-1190
[17]Verheijen R, Kuijpers HJ. Sehlingemann R, et al. Ki-67 detects a nuclear matrix-associated antigen. Localization in mitotic cells and association withchromosomes[J], J Cell Sci,1989,92(5):531-540
[18]Khalifeh I, Munkarah A R, Schimp V, et al. The impact of c-kit and Ki-67 expression on patients prognosis in advanced ovarian serouscarcinoma. Int JGynecol Pathol,2005,24:228-234
[19]Bruno S, Darzynkiewicz ZP, Van Diest PJ, et al. Cell cycle dependent expression and stability of the nuclear protein detected hy Ki-67 antibody in HL-60cell[J]. CellProlif.1992,25(1):31-40
[20]Miyazono K, Suzuki H, Imamura T. Regulation of TGF2beta signalingand its roles in progression of tumors.[J]. Cancer Sci 2003,94(3):230-234
[21]Balch CM, Buzaid AC, Atkins MB, et al.A new American joint committee on cancer staging system for cutaneous melanoma. Cancer,2000,88:1484-1491
[22]HugBA, Ahmed N, Robbins JA, et al. A chromatin immunop recip ita2tion screen reveals p rotein kinase Cbeta as a direct RUNX1 targetgene. [J].J Biol Chem,2004,279 (2):825-830
[23]Ito Y, Miyazono K. Runx transcrip tion factors as key targets of TGF2beta superfamily signaling[J]. CurrOp in Genet Dev,2003,13(1):43-47
[24]Zhang YW, Yasui N, Ito K, et al. A Runx2/PEBP2 alpha A/CB2FA1 mutation disp laying impaired transactivation and Smad interac2tion in cleidocranial dysp lasia[J]. Proc NatlAcad SciUSA,2000,97(19):10549-10554
[25]Bae SC, Choi JK. Tumor suppressor activity of Runx3.[J]. Onco gene,2004,23(24):4336-4340
[26]Bangsow C, Rubins N, Glusman G, et al. The Runx3 gene2se2quence,structure and regulated exp ression[J].Gene 2001,279(2):221-232
[27]LiQL, Ito K, Sakaura C, et al. Causal relationship between the loss of Runx3 expression and gastric cancer[J]. Cell,2002,109(1):113-124
[28]Groneberg DA, Witt H, Adcock IM, et al. Smads as intracellularmediators of airway inflammation[J]. Exp Lung Res,2004,30(3):223
[29]IJiu F, Pouponnot C, Massague J. Dual role of the Smad4/DPC4tumor suppressor in TGF-βinducible transcriptional complex [J]. Genes Dev.1997.11(23):3157
[30]Shi Y, Massague J. Mechanisms of TGF-P signaling from cell men-brane to the nucleus[J]. Cell,2003,113(6):685
[31]Dijke P, Hill CS, New insiights into TGF-β/Smadsignaling[J]. Trends Biochemical Sci.2004.29(5):265
[32]Derynck R, Zhang YE. Smad-depended and Smad-independedpathways in TGF-β family signaling[J]. Nature,2003,425(6958):577
[33]Shi Y, Massague J. Mechanisms of TGF-β signaling from cell menbrane to the nucleus[J]. Cell,2003,113(6):685
[34]Xu L, Kang Y, Col S, et al. Smad2 nucleocy to plasmic shuttling by nucleoporins CAN/ Nup214 and Nup153 feeds TGF-P signaling complexes in the cytoplasm and nucleus[J]. Mol Cell,2002,10(2):271
[35]Inman GJ, Nicolas FJ, Hil CS. Nucleocytoplasmic shuttling ofSmad2,3 and 4 permits sensing of TGF-β receptor activity[J]. Mol Cell,2002,10(2):283
[36]Reinacher-Schick A, Baldus SE, Romdhana B, et al. Loss of Smad4 correlates with loss of the invasion suppressor E-cadherin in advanced colorectal carcinomas[J].J Pathol, 2004.202(4):412
[37]Volmer MW, Radacz Y, Hahn SA, et al. Tumor suppressor Smad4mediates dowrrregn lation of the anti-adhesive invasion-promoting matricellular protein SPARC:Landscaping activity of Smad4 as revealed by asecretome analysis[J]. Proteomics,2004,4(5):1324
[38]Schwarte-Waldhof I, Volpert OV, Bouck NP, et al.Smad4/DPC4-mediated tumor suppression through suppression of angiogenesis[J]. Proc Natl Acad Sci USA,2000,97(17):9624
[39]刘鹏熙,陈前军,钟少文,等.新辅助化疗对乳腺癌P170、Ki67、p53的影响[J].肿瘤.2006,26(2)：168
[40]邱吉庆,闫世军,赵刚,等.Ki-67表达对人脑胶质瘤恶性度的评价[J].中风与神经疾病杂志,2001,18(2)：81-83
[41]Porter, PL, Malone. KE, Heagerty PJ. et al. Expression of cell-cycle Regulators P27kiP1 and cyclin E. Alone and in combination,combination withSurvival in young breast cancer patients [J]. Natt Med,1997,3(2):222-225
[42]Cardone, MH. Roy N. Stenniche HR, et al. Regulation of cell deathProtease caspase 9 byphasphory-lation [J]. Scienee,1998,282:1318-1325
[43]Evan GI, Vousden KH. Proliferation, cell cycle and apoptosis in cancer[J]. Nature,200 1,411(4):342
[44]Sherr CJ. The Pezcoller Lecture:Cancer cell cyclerevisited[J]. Cancer Res,2000.60(29):3689
[45]詹启敏.分子肿瘤学[M].第1版.北京：人民卫生出版社,2005：25-39
[1]Ries LAG, Eisner MP, Kosary, et al editors. SEER Cancer Statistics Re VIEW,1973-2001 Bethesda,MD.NationalCancerInstitute,2004.Availablefrom:http://seer,cancer.gov/csr/1975-2001. Accessed May,2004
[2]Slominski A, Wortsman J, Carlson AJ, et al. Malignant melanoma [J]. Arch Pathol LabMed, 2001,125(10):1295-306
[3]王娜,陆洪光.黑色素瘤相关基因的研究新进展.国外医学肿瘤学分册,2004,31(增)：43-47
[4]刘巍峰,牛晓辉.黑色素瘤发病机制初步探讨.中国骨肿瘤骨病,2007,5(6)：294
[5]Yogeeswaran G, Gronberg A, Hansson M, et al. Correlation ofglycosphingolipids and sialicacid in Yac-1 lymphoma variants withtheirsensitivitytonaturalkill-cell-mediatedlysis. IntJCancer,1981, 28:517
[6]Bangsow C, Rubins N, Glusman G, et al. The Runx3 gene-sequence, structure and regulated expression [J]. Gene 2001,279 (2):221-232
[7]Kato N, Tamura G, Fukase M, et al. Hypermethylation of the Runx3 gene promoter in testicular yolk sactumor of infants [J]. Am JP athol,2003,163(2):387-391
[8]Coffman JA. Runx transcription factors and the developmental balance between cell proliferation and differentiation [J]. Cell Biol Int2003,27(4):315-324
[9]Tang YY, Shi J, Zhang L, et al. Energetic and functional contributionof residues in the core binding factor beta (CBFbeta) subunit to heterodimerization with CBFalpha [J]. J Biol Chem 2000,275(50):39579-39588
[10]Yang N, Zhang L, Zhang Y, et al. An important role for Runx3 inhuman L1 transcription and retro transposition [J]. Nucleic Acids Res2003,31(16):4929-4940
[11]Ito Y. Molecular basis of tissue-specific gene expression mediated bythe runt domain transcription factor PEBP2/CBF[J]. Genes Cells1999,4(12):685-696
[13]Lund AH, van Lohuizen M. Runx atrilogy of cancer genes [J]. Cancer Cell 2002,1(3):213-215
[14]HugBA, Ahmed N, Robbins JA, et al. A chromatin immunoprecipitation screen reveals protein kinase Cbeta as a direct Runxl targetgene[J]. J Biol Chem,2004,279(2):825-830
[15]Ito Y, Miyazono K. Runx transcription factors as key targets of TGF-beta super family signaling[J]. Curropin Genet Dev,2003,13(1):43-47
[16]Zhang YW, Yasui N, Ito K, et al. A Runx2/PEBP2 alpha A/CB-FA1 mutation disp laying impaired transactivation and Smad interaction in cleidocranial dysplasia[J]. Proc NatlAcad SciUSA,2000,97(19):10549-10554
[17]Bae SC, Choi JK. Tumor supp ressoractivity of Runx3[J]. Onco-gene,2004,23(24):4336-4340
[18]Miyazono K, Suzuki H, Imamura T. Regulation of TGF-beta signalingand its roles in progression of tumors[J]. Cancer Sci 2003,94(3):230-234
[19]Miyazono K, Suzuki H, Imamura T. Regulation of TGF-beta signal in and its roles in progression of tumors[J]. Cancer Sci,2003,94:230-234
[20]Zaidi SK, Sullivon AJ, Wijnen AJV, et al. Integration of Runx and Smad regulatory signals at transcriptionally active subnuclear sites[J]. Proc Natl Acad Sci USA,2002,99(12):8048-8053
[21]LiQL, Ito K, Sakaura C, et al.Causal relationship between the loss ofRunx3 expression and gastric Cancer[J]. Cell,2002,109(1):113-124
[22]Miyazono K, Suzuki H, Imamura etal. Regulation of TGF-beta signaling and itsroles in progssion oftumors.Cancer Sci.2003,94(31):230-234
[23]Li E. Chromatin modification and epigenetic reprogramming in mammalian development[J]. Nat Rev Genet,2003,3:662-673
[24]Ehrlich M. The controversial denouement of vertebrate DNA methylation research[J]. Biochemistry,2005,70:568-575
[25]Courtier B, Heard E, Avner P, et al. Xce hap lotypes show modi2fied methylation in a region of the active X chromosome lying-3'to Xist[J]. Procnatl Acad Sci USA,1995,92:3531-3535
[26]Momparler RL, Bovenzi V. DNA methylation and cancer[J].J Cell Physiol,2000,183:145-154
[27]Li Q L, Ito K, Sakakura C, et al. Causal relatiaonship between the loss of Runx3 expression and gastric cancer [J]. Cell,2002,109(1):113-124
[28]Guo WH, Weng LQ, Ito K, et al.Inhibition of growth of mouse gastric cancer cells by Runx3, a novel tumor suppressor[J]. Onco-gene.2002,21:8351-8355
[29]Wei D, Gong W, Oh SC, et al.Loss of Runx3 expression significantly affect the clinical outcome of gastric cancer patients and its restoration causes drastic suppression of tumor growth and metastasis [J]. Cancer Res,2005,65(11):4809-4816
[30]Waki T, Tamura G, Sato M. et aL Promoter methylation status of DAP kinaseand Runx3 genes in neoplastic and nonneoplastic gastric epithelia CancerSci.2003,94(4):360-364
[31]Li QL, Kim HR, Kim WJ, et al. Transcriptional silencing of the Runx3 gene by CpG hyper methyl lation is associated with lung cancer. BiochemBiophysResCommun.2004,314(1):223-228
[32]Kato N, Tamura G, Fu kase M, et al. Hypermethylation of the Runx3 genepromoter in testicular yolk 8ac tumor of infants. Am J Patho 1.2003,163(2)387-391
[33]Wada M, Yasumi S, Takaishi S, et al. Frequent loss of Runx3 gene expression in human bile duct and pancreatic cancer ceil lines. Oncogene.2004,23(13):2401-2407
[35]Xiao WH, Liu WW. Hemizygous deletion and hypermethylation of Runx3 genein hepato cellular carcinoma. World J Gastroenteml,2004,10(3):376-380
[36]Ku J L, Kang S B, Shin YK. et al. Promoter hypermethylation downregulatesRunx3 gene expression in colorectal cancer cell lines[J]. Oncogene,2004,23(40):6736-6742
[37]Kang GH, Lee S, et al. Aberrant CpG island hypermethylation of multiple genesin prostate cancer and prostatic intraepithelial neoplasia[J]. Path01.2002(2):233-240
[38]Kim TY, Lee HJ, Hwang KS, et al. Methylation of Runx3 in various typesof human cancers and premalignant stages of gastric carcinoma[J]. Lab Invest,2004,84(4):479-484
[39]Salto. Tellez M, Peh BK, Ito K, Tan SH et al. Runx3 protein is overexpressed inhuman basal cell carcinomas.Oncogene.2006 Dec 7;25(58):7646-9
[2]RiesLAG,EisnerMP,Kosary,etaleditors.SEERCancerStatisticsREVIEW,1973-2001.Bethesda, MD.NationalCancerInstitute,2004.Availablefrom:http://seer,cancer.gov/csr/1975-2001.Accesses May,2004
[3]Kang GH, Lee S, Lee HJ, Hwang KS. Aberrant CpG island hypermethyaltion ofmultiple genes in prostatic cacenr and prostatic intraepithelial neoplasia [J]. J Pathol 2004;202:233-240
[4]Kjm TY, Lee HJ, Hwang KS, Lee M.Kim JW, Bang YJ, Kang GH. Methylation of Runx3 in various types of human cancers premalignant, stages of gastric carcinoma [J] Lab Invest 2004;84:479-484
[5]Wada M, Yazumi S, Takaishi S, Hasegawa K, Sawada M, Tanaka H, Ida H, SakakuraC, Ito Y, Chiba T. Frequent loss of Runx3 gene expression in human bile duct andPancreatic cancer cell lincs [J]. Oncogene 2004;23:2401-2407
[6]Li QL, Kim WJ, Choi JK, Lee YH, Kim HM, Li LS, Kim H Chang J, ItoY, Youl LeeK.Bae SC. TranscriPtional silencing of the Runx3 gene by CpG hypermethylation isAssociated with lung cancer [J]. Biochem Biophys Res Commun 2004;314:223-228
[7]Schulze-Bergkamen H, Krammer PH. Apoptosis in cancer implication for therapy [J]. Semin Oncol 2004;31:90-119
[8]Li QL, Ito K, Sakakura C, et al. Causal relationship between the loss of Runx3 expression and gastric cancer [J].Cell,2002,109(1):113-124
[9]Baylin SB, Belinsky SA, Herman JG. Aberrant methylation of gene promoters inCancer concepts, misconcepts and, Promise [J].JNatl Cancer, Inst,2000,92:1406-1461
[10]Ito Y, Miyazono K. Runx3 transcription factors as key targets of TGF-beta superfamily signaling [J]. Currop in GenetDev,2003,13:34-47
[11]Zaidi SK, Sullivan AJ, van Wijnen AJ, et al. Integration of Runx and smad regulatorySignals at tannscriptionally active subnuclear sites [J] ProcNatlAcad SciUSA,2002,99(12):8048-8053
[12]Kato N, Tamura G, Fukase M, et al.HyPermethylation of the Runx3 gene Promoterin testicular yolk sactumor of infants [J], Am J Pathol,2003,163(2):387-398
[13]Schwarte-Waldhoff I, Schmiegel W. Smad4 transcriptional path-ways and angiogenesis[J]. Int J Gastrointest Cancer,2002,31(1):47-59
[14]Schwarte-Waldhoff 1, Klein S, Blass-Kampmann S, et al. DPC4/Smad4 mediated tumor supp-ression of colon carcinorma cells is associated with reduced urokinses expression[J]. Oncogene,1999,18(26):3152-3158
[15]Asakura T, Takano Y, Iki M, et al. Prognostic value of Ki-67 for recurrence andprogression of superficial bladder cancer[J]. J Urol,1997,158(2):385-388
[16]孙妍,曹玉文,陆天才,等.宫颈上皮癌变过程中细胞分化的研究[J].癌症,2005,24(10)：1184-1190
[17]Verheijen R, Kuijpers HJ. Sehlingemann R, et al. Ki-67 detects a nuclear matrix-associated antigen. Localization in mitotic cells and association withchromosomes[J], J Cell Sci,1989,92(5):531-540
[18]Khalifeh I, Munkarah A R, Schimp V, et al. The impact of c-kit and Ki-67 expression on patients prognosis in advanced ovarian serouscarcinoma. Int JGynecol Pathol,2005,24:228-234
[19]Bruno S, Darzynkiewicz ZP, Van Diest PJ, et al. Cell cycle dependent expression and stability of the nuclear protein detected hy Ki-67 antibody in HL-60cell[J]. CellProlif.1992,25(1):31-40
[20]Miyazono K, Suzuki H, Imamura T. Regulation of TGF2beta signalingand its roles in progression of tumors.[J]. Cancer Sci 2003,94(3):230-234
[21]Balch CM, Buzaid AC, Atkins MB, et al.A new American joint committee on cancer staging system for cutaneous melanoma. Cancer,2000,88:1484-1491
[22]HugBA, Ahmed N, Robbins JA, et al. A chromatin immunop recip ita2tion screen reveals p rotein kinase Cbeta as a direct RUNX1 targetgene. [J].J Biol Chem,2004,279 (2):825-830
[23]Ito Y, Miyazono K. Runx transcrip tion factors as key targets of TGF2beta superfamily signaling[J]. CurrOp in Genet Dev,2003,13(1):43-47
[24]Zhang YW, Yasui N, Ito K, et al. A Runx2/PEBP2 alpha A/CB2FA1 mutation disp laying impaired transactivation and Smad interac2tion in cleidocranial dysp lasia[J]. Proc NatlAcad SciUSA,2000,97(19):10549-10554
[25]Bae SC, Choi JK. Tumor suppressor activity of Runx3.[J]. Onco gene,2004,23(24):4336-4340
[26]Bangsow C, Rubins N, Glusman G, et al. The Runx3 gene2se2quence,structure and regulated exp ression[J].Gene 2001,279(2):221-232
[27]LiQL, Ito K, Sakaura C, et al. Causal relationship between the loss of Runx3 expression and gastric cancer[J]. Cell,2002,109(1):113-124
[28]Groneberg DA, Witt H, Adcock IM, et al. Smads as intracellularmediators of airway inflammation[J]. Exp Lung Res,2004,30(3):223
[29]IJiu F, Pouponnot C, Massague J. Dual role of the Smad4/DPC4tumor suppressor in TGF-βinducible transcriptional complex [J]. Genes Dev.1997.11(23):3157
[30]Shi Y, Massague J. Mechanisms of TGF-P signaling from cell men-brane to the nucleus[J]. Cell,2003,113(6):685
[31]Dijke P, Hill CS, New insiights into TGF-β/Smadsignaling[J]. Trends Biochemical Sci.2004.29(5):265
[32]Derynck R, Zhang YE. Smad-depended and Smad-independedpathways in TGF-β family signaling[J]. Nature,2003,425(6958):577
[33]Shi Y, Massague J. Mechanisms of TGF-β signaling from cell menbrane to the nucleus[J]. Cell,2003,113(6):685
[34]Xu L, Kang Y, Col S, et al. Smad2 nucleocy to plasmic shuttling by nucleoporins CAN/ Nup214 and Nup153 feeds TGF-P signaling complexes in the cytoplasm and nucleus[J]. Mol Cell,2002,10(2):271
[35]Inman GJ, Nicolas FJ, Hil CS. Nucleocytoplasmic shuttling ofSmad2,3 and 4 permits sensing of TGF-β receptor activity[J]. Mol Cell,2002,10(2):283
[36]Reinacher-Schick A, Baldus SE, Romdhana B, et al. Loss of Smad4 correlates with loss of the invasion suppressor E-cadherin in advanced colorectal carcinomas[J].J Pathol, 2004.202(4):412
[37]Volmer MW, Radacz Y, Hahn SA, et al. Tumor suppressor Smad4mediates dowrrregn lation of the anti-adhesive invasion-promoting matricellular protein SPARC:Landscaping activity of Smad4 as revealed by asecretome analysis[J]. Proteomics,2004,4(5):1324
[38]Schwarte-Waldhof I, Volpert OV, Bouck NP, et al.Smad4/DPC4-mediated tumor suppression through suppression of angiogenesis[J]. Proc Natl Acad Sci USA,2000,97(17):9624
[39]刘鹏熙,陈前军,钟少文,等.新辅助化疗对乳腺癌P170、Ki67、p53的影响[J].肿瘤.2006,26(2)：168
[40]邱吉庆,闫世军,赵刚,等.Ki-67表达对人脑胶质瘤恶性度的评价[J].中风与神经疾病杂志,2001,18(2)：81-83
[41]Porter, PL, Malone. KE, Heagerty PJ. et al. Expression of cell-cycle Regulators P27kiP1 and cyclin E. Alone and in combination,combination withSurvival in young breast cancer patients [J]. Natt Med,1997,3(2):222-225
[42]Cardone, MH. Roy N. Stenniche HR, et al. Regulation of cell deathProtease caspase 9 byphasphory-lation [J]. Scienee,1998,282:1318-1325
[43]Evan GI, Vousden KH. Proliferation, cell cycle and apoptosis in cancer[J]. Nature,200 1,411(4):342
[44]Sherr CJ. The Pezcoller Lecture:Cancer cell cyclerevisited[J]. Cancer Res,2000.60(29):3689
[45]詹启敏.分子肿瘤学[M].第1版.北京：人民卫生出版社,2005：25-39
[1]Ries LAG, Eisner MP, Kosary, et al editors. SEER Cancer Statistics Re VIEW,1973-2001 Bethesda,MD.NationalCancerInstitute,2004.Availablefrom:http://seer,cancer.gov/csr/1975-2001. Accessed May,2004
[2]Slominski A, Wortsman J, Carlson AJ, et al. Malignant melanoma [J]. Arch Pathol LabMed, 2001,125(10):1295-306
[3]王娜,陆洪光.黑色素瘤相关基因的研究新进展.国外医学肿瘤学分册,2004,31(增)：43-47
[4]刘巍峰,牛晓辉.黑色素瘤发病机制初步探讨.中国骨肿瘤骨病,2007,5(6)：294
[5]Yogeeswaran G, Gronberg A, Hansson M, et al. Correlation ofglycosphingolipids and sialicacid in Yac-1 lymphoma variants withtheirsensitivitytonaturalkill-cell-mediatedlysis. IntJCancer,1981, 28:517
[6]Bangsow C, Rubins N, Glusman G, et al. The Runx3 gene-sequence, structure and regulated expression [J]. Gene 2001,279 (2):221-232
[7]Kato N, Tamura G, Fukase M, et al. Hypermethylation of the Runx3 gene promoter in testicular yolk sactumor of infants [J]. Am JP athol,2003,163(2):387-391
[8]Coffman JA. Runx transcription factors and the developmental balance between cell proliferation and differentiation [J]. Cell Biol Int2003,27(4):315-324
[9]Tang YY, Shi J, Zhang L, et al. Energetic and functional contributionof residues in the core binding factor beta (CBFbeta) subunit to heterodimerization with CBFalpha [J]. J Biol Chem 2000,275(50):39579-39588
[10]Yang N, Zhang L, Zhang Y, et al. An important role for Runx3 inhuman L1 transcription and retro transposition [J]. Nucleic Acids Res2003,31(16):4929-4940
[11]Ito Y. Molecular basis of tissue-specific gene expression mediated bythe runt domain transcription factor PEBP2/CBF[J]. Genes Cells1999,4(12):685-696
[13]Lund AH, van Lohuizen M. Runx atrilogy of cancer genes [J]. Cancer Cell 2002,1(3):213-215
[14]HugBA, Ahmed N, Robbins JA, et al. A chromatin immunoprecipitation screen reveals protein kinase Cbeta as a direct Runxl targetgene[J]. J Biol Chem,2004,279(2):825-830
[15]Ito Y, Miyazono K. Runx transcription factors as key targets of TGF-beta super family signaling[J]. Curropin Genet Dev,2003,13(1):43-47
[16]Zhang YW, Yasui N, Ito K, et al. A Runx2/PEBP2 alpha A/CB-FA1 mutation disp laying impaired transactivation and Smad interaction in cleidocranial dysplasia[J]. Proc NatlAcad SciUSA,2000,97(19):10549-10554
[17]Bae SC, Choi JK. Tumor supp ressoractivity of Runx3[J]. Onco-gene,2004,23(24):4336-4340
[18]Miyazono K, Suzuki H, Imamura T. Regulation of TGF-beta signalingand its roles in progression of tumors[J]. Cancer Sci 2003,94(3):230-234
[19]Miyazono K, Suzuki H, Imamura T. Regulation of TGF-beta signal in and its roles in progression of tumors[J]. Cancer Sci,2003,94:230-234
[20]Zaidi SK, Sullivon AJ, Wijnen AJV, et al. Integration of Runx and Smad regulatory signals at transcriptionally active subnuclear sites[J]. Proc Natl Acad Sci USA,2002,99(12):8048-8053
[21]LiQL, Ito K, Sakaura C, et al.Causal relationship between the loss ofRunx3 expression and gastric Cancer[J]. Cell,2002,109(1):113-124
[22]Miyazono K, Suzuki H, Imamura etal. Regulation of TGF-beta signaling and itsroles in progssion oftumors.Cancer Sci.2003,94(31):230-234
[23]Li E. Chromatin modification and epigenetic reprogramming in mammalian development[J]. Nat Rev Genet,2003,3:662-673
[24]Ehrlich M. The controversial denouement of vertebrate DNA methylation research[J]. Biochemistry,2005,70:568-575
[25]Courtier B, Heard E, Avner P, et al. Xce hap lotypes show modi2fied methylation in a region of the active X chromosome lying-3'to Xist[J]. Procnatl Acad Sci USA,1995,92:3531-3535
[26]Momparler RL, Bovenzi V. DNA methylation and cancer[J].J Cell Physiol,2000,183:145-154
[27]Li Q L, Ito K, Sakakura C, et al. Causal relatiaonship between the loss of Runx3 expression and gastric cancer [J]. Cell,2002,109(1):113-124
[28]Guo WH, Weng LQ, Ito K, et al.Inhibition of growth of mouse gastric cancer cells by Runx3, a novel tumor suppressor[J]. Onco-gene.2002,21:8351-8355
[29]Wei D, Gong W, Oh SC, et al.Loss of Runx3 expression significantly affect the clinical outcome of gastric cancer patients and its restoration causes drastic suppression of tumor growth and metastasis [J]. Cancer Res,2005,65(11):4809-4816
[30]Waki T, Tamura G, Sato M. et aL Promoter methylation status of DAP kinaseand Runx3 genes in neoplastic and nonneoplastic gastric epithelia CancerSci.2003,94(4):360-364
[31]Li QL, Kim HR, Kim WJ, et al. Transcriptional silencing of the Runx3 gene by CpG hyper methyl lation is associated with lung cancer. BiochemBiophysResCommun.2004,314(1):223-228
[32]Kato N, Tamura G, Fu kase M, et al. Hypermethylation of the Runx3 genepromoter in testicular yolk 8ac tumor of infants. Am J Patho 1.2003,163(2)387-391
[33]Wada M, Yasumi S, Takaishi S, et al. Frequent loss of Runx3 gene expression in human bile duct and pancreatic cancer ceil lines. Oncogene.2004,23(13):2401-2407
[35]Xiao WH, Liu WW. Hemizygous deletion and hypermethylation of Runx3 genein hepato cellular carcinoma. World J Gastroenteml,2004,10(3):376-380
[36]Ku J L, Kang S B, Shin YK. et al. Promoter hypermethylation downregulatesRunx3 gene expression in colorectal cancer cell lines[J]. Oncogene,2004,23(40):6736-6742
[37]Kang GH, Lee S, et al. Aberrant CpG island hypermethylation of multiple genesin prostate cancer and prostatic intraepithelial neoplasia[J]. Path01.2002(2):233-240
[38]Kim TY, Lee HJ, Hwang KS, et al. Methylation of Runx3 in various typesof human cancers and premalignant stages of gastric carcinoma[J]. Lab Invest,2004,84(4):479-484
[39]Salto. Tellez M, Peh BK, Ito K, Tan SH et al. Runx3 protein is overexpressed inhuman basal cell carcinomas.Oncogene.2006 Dec 7;25(58):7646-9