摘要
背景与目的
肺癌是目前世界上最普遍的恶性肿瘤之一,其发病率和病死率均不断上升,因肺癌死亡人数已跃居癌症死亡总人数的首位。从确诊到死亡,其五年生存率不到15%,造成肺癌高死亡率的主要原因是缺乏有效的早期诊断方法,因此寻找新的有效的早期诊断方法是目前肺癌研究领域的热点。肿瘤相关抗原(tumor-associated antigen,TAA)抗体的检测做为一种新的肺癌早期诊断方法已成为肺癌诊断领域的热点,但是单一抗原抗体的检测敏感性和特异性都比较差,用于肺癌诊断有一定的局限性。为此,本研究对15种肿瘤相关抗原(Koc、p62、CyclinE、CyclinB1、Imp1、p16、Sui1、Survivin、Calnuc、C-myc、p53、CyclinD1、CDK2、cIAP1和RalA)抗体在肺癌患者血清中的表达水平进行检测,以评价其在肺癌诊断中的价值与临床意义。
材料与方法
1.样本采集:研究对象共266例,其中肺癌患者:174例,其中男性109例,女性65例,年龄35～84岁,平均年龄62.6±11.7岁,均经病理证实为肺癌;健康对照92例,其中男性38例,女性54例,年龄19～76岁,平均50.6±15.1岁,均为健康体检并被证实无任何肿瘤相关疾患的健康人。
2.试验方法:用间接酶联免疫吸附实验(enzyme-linked-immunosorbentserologic assay,ELISA)检测每个抗原相应抗体在肺癌患者中的表达水平;用狭缝印迹杂交(slot blot hybridization)实验对ELISA阳性结果做进一步的验证。
3.统计学处理:所有数据采用SPSS13.0统计分析软件包进行处理,统计分析采用χ~2检验,当四格表中有理论数小于1或N＜40时,用四格表确切概率法,检验水准为α=0.05。
结果
1.肺癌组和健康对照组15种TAAs抗体检测的阳性率:p16分别为25.9%、2.2%;RalA分别为14.4%、3.3%;Koc分别为9.8%、2.2%;CyclinD1分别为12.1%、2.2%;p62分别为17.2%、3.3%;Sui1分别为16.1%、4.3%;cIAP1分别为12.6%、0%;CyclinE分别为21.3%、3.3%;Calnuc分别为27.0%、2.2%;Imp1分别为21.3%、1.1%;Survivin分别为14.9%、1.1%;p53分别为24.1%、3.3%;C-myc分别为22.4%、0%;CyclinB1分别为16.1%、3.3%;CDK2分别为11.5%、3.3%。
2.所有抗原抗体(Koc、Sui1、Survivin、p62、CyclinE、Imp1、p16、Calnuc、C-myc、p53、CyclinD1、CyclinB1、CDK2、cIAP1和RalA)在肺癌患者血清中的的阳性率均显著高于健康对照组,差异有统计学意义(P＜0.05)。
3.肺癌组p62和p16的抗体阳性率与肺癌病理类型有关,其阳性率均以肺鳞癌最高,分别为48.3%、27.6%。与其它病理类型相比,p16抗体在肺鳞癌中的表达高于肺腺癌和小细胞肺癌(P＜0.05);p62抗体在肺鳞癌中的表达高于小细胞肺癌(P＜0.05)。
4.15种抗原抗体联合检测的灵敏度为73.0%,特异度为90.2%,约登指数为0.632,阳性预测值为93.4%,阴性预测值为63.8%,阳性似然比为7.45,一致率为78.9%,Kappa为0.58。
结论
1.Koc、Sui1、Survivin、p62、CyclinE、Imp1、p16、Calnuc、C-myc、p53、CyclinD1、CyclinB1、CDK2、cIAP1和RalA15种TAAs抗体在肺癌患者中表达较高,在健康对照者中表达相对较低,有助于肺癌的诊断。
2.p16、p62对肺癌的诊断均有一定价值,且p16在肺鳞癌中的表达高于肺腺癌和小细胞肺癌;p62在肺鳞癌中的表达高于小细胞肺癌,提示其有助于肺癌的病理分型。
3.15种TAA抗体的表达水平在肺癌患者中均不存在性别差异;除RalA、Calnuc外,其余13种TAA抗体的表达水平在肺癌患者中均不存在年龄差异。
4.15种TAA抗体联合检测可提高肺癌的检出率,有助于肺癌的诊断。
Background and Objective
Lung cancer is one of the most common malignancies,both the incidence rate and mortality rate of lung cancer have been on the rise,with its mortality rate ranking the first among various kinds of malignancies.The majority of lung cancers remain clinically undetectable until patients have developed to the advanced-stage disease, and the five-year survival rate of lung cancer only accounts for less than 15%.The main reason for such a high mortality rate is that lung cancer is difficult to be diagnosed at the early stage.Therefore,it is crucial to search a useful method which can diagnose lung cancer at the early stage.TAA(tumor-associated antigen) is one of the major research topics in modern oncology,while the TAA detection of lung cancer has become an important target for the diagnosis of the malignancy.But TAA shows its limitations in the diagnosis of lung cancer,and would not satisfactory in the sensitivity and specificity.Therefore,we have used a multiple TAAs including Koc, p62,CyclinE,CyclinB1,Imp1,p16,Suil,Survivin,Calnuc,C-myc,p53,CyclinD1, CDK2,cIAP1 or RalA in detection anti-TAA antibodies for the diagnosis of lung cancer.The aim of this study is to evaluate the diagnostic value of a TAA array in detection of anti-TAA antibodies for the diagnosis of lung cancer.
Materials and Methods
1.Sample preparation:in this study,all patients were divided into two groups: lung cancer group consisted by 174 cases,109 males and 65 females,age from 35 to 84 years,mean age 62.6±11.7 years,all cases had been proved by pathologic diagnosis;health comparison group consisted by 92 cases,38 males and 54 females, age from 19 to 76years,mean age 50.6±15.1 years.
2.The enzyme-linked immunoassay assay(ELISA) was performed to detect all TAAs antibodies,and the slot blot was used to checking the results of the ELISA.
3.Statistical analysis:all date were analyzed by SPSS 13.0.In the statistics analysis,χ~2 test and the exact probability of fourfold table were used;size of the testα=0.05.
Results
1.The positive gender rate of the fifteen TAA,which was detected in this study, in lung cancer and health comparison group:p16 is 25.9%,2.2%,respectively;RalA is 14.4%,3.3%,respectively;Koc is 9.8%,2.2%,respectively;CyclinD1 is 12.1%, 2.2%,respectively;p62 is 17.2%,3.3%,respectively;Suil is 16.1%,4.3%, respectively;cIAP1 is 12.6%,0%,respectively;CyclinE is 21.3%,3.3%,respectively; Calnuc is 27.0%,2.2%,respectively;Imp1 is 21.3%,1.1%,respectively;Survivin is 14.9%,1.1%,respectively;p53 is 24.1%,3.3%,respectively;C-myc is 22.4%,0%, respectively;CyclinB1 is 16.1%,3.3%,respectively;CDK2 is 11.5%,3.3%, respectively.
2.In serum samples from patients with lung cancer,the positive gender rate of Koc,Suil,Survivin,p62,CyclinE,Imp1,p16,Calnuc,C-myc,p53,CyclinD1, CyclinB1,CDK2,cIAP1 and RalA is remarkably higher than that in the health comparison group,the difference has statistical significance(P＜0.05).
3.The positive gender rate of p62 and p16 in the lung cancer group have relation with the pathology type of lung cancer.The highest pathology type in p62 and p16 is squamous cell carcinoma,respectively is 48.3%,27.6%.Compared with other pathological types,the difference has statistica significance(P＜0.05).
4.The sensitivity,specificity,Youden's index,positive predictive value,negative predictive value,positive likelihood ratio,agreement rate value and Kappa of the 15 anti-TAA antibodies in detection of lung cancer is 73.0%,90.2%,0.632,93.4%, 63.8%,7.45,78.9%,0.58,respectively.
Conclusions
1.The expression of Koc,Suil,Survivin,p62,CyclinE,Imp1,p16,Calnuc, C-myc,p53,CyclinD1,CyclinB1,CDK2,cIAP1 and the RalA in lung cancer group is higher than the health group.This is useful to diagnose lung cancer.
2.p16 and p62 have certain value to the lung cancer diagnosis,and the positive gender rate in the squamous cell carcinoma is higher than that in the adenocarcinoma. This result indicates that the detection of p16 and p62 is useful to judge the pathology type of lung cancer.
3.There is no statistical significance in all TAAs which were detected in this study between different sex groups;except RalA and Calnuc,all other TAAs all do not have statistical significance between different age groups.
4.The combinational application of 15 TAAs could be a valuable method.This approach can raise the sensitivity rate in serological diagnosis of lung cancer.
引文
1.Haugen A.Molecular biology in diagnosis of lung cancer[J].Tidsskr Nor Laegeforen,2005,125(23):3283-3285.
2.World Health Organization.Gender in lung cancer and smoking research JR]In:Lung cancer.WHO Library Cataloguing-in-Publication Data,2005,5-8.
3.Gran ville Ca,Dennis PA.An overview of lung cancer genomics and proteomics [J].Am J Respir Cell Mol Biol,2005,32(3):169-176.
4.李龙芸.继续深入开展肺癌的基础与临床研究[J].中华内科杂志,2000,39(11):725-726.
5.陈娟,张锦.肺癌早期诊断的研究进展[J].肿瘤研究与临床,2003,15(2):134-137.
6.Strauss GM,Skarin AT.Use of tumor markers in lung cancer[J].Hematol Oncol Clin North Am.1994,8(3):507-532.
7.Roigas J,Schulze G,Raytarowski S,et al.Tumor M2 pyruvate kinase in renal cell carcinoma.Studies of plasma inpatients[J].Urologe A,2000,39(6):554-556.
8.Kim CW,Kim JI,Park SH,et al.usefulness of plasma tumor M2-pyruvate kinase in the diagnosis of gastrointestinal cancer[J].Korean J Gastroenterol,2003,42(5):387-393.
9.Zhang JY.Tumor-associated antigen arrays to enhance antibody detection for cancer diagnosis[J].Cancer Detection and prevention,2004(28):114-118.
10.王鸿程,许学亮,简丽萍,等.STAT5与CyclinB1、CyclinD1、c-fos、C-myc 在非小细胞肺癌中的表达及临床意义[J].泸州医学院学报,2008,31(6):601-605.
11.徐克.肺癌遗传学与分子生物学最新进展[J].中国肺癌杂志,第十二届世界肺癌大会专题,2008,11(1):29-33.
12.Frusch N,Bosquee L,Louis R.lung cancer.Epidemiology and etiologic factors [J].Rev Med Liege,2007,62(9):548-553.
13.Notkins A,Lernmark A.Autoimmune typeldiabetes:resolved and unresolved issues[J].J Clin Invest,2001,108(9):1247-1252.
14.Rapoport B,McLachlan SM.Thyroid autoimmunity[J].J Clin Invest 2001,108(9):1253-1259.
15.Megliorino R,Shi FD,Peng XX,et al.Autoimmune response to anti-apoptotic protein survinvin and its association with antibodies to p53 and C-myc in cancer detection[J].Cancer Detect Prev,2005,29(3):241-248.
16.Shi FD,Zhang JY,Liu D,et al Preferential Humoral Immune Response in Prostate Cancer to Cellular Proteins p90 and p62 in a Panel of Tumor-Associated Antigens[J].Prostate,2005,63(3):252-258.
17.Zhang JY.Mini-array of tumor-associated antigen to enhance autoantibody detection for immunodiagnosis of hepatocellular carcinoma[J].Autoimmun Rev,2007,6(3):143-148.
18.Tan EM,Zhang JY.Autoantibodies to tumor-associated antigens:reporters from the immune system[J].Immunol Rev,2008(222):328-340.
19.Zhang JY,Edward KL,Peng XX,et al.A novel cytoplastic protein with RNA-binding motifs is an autoantigen in human hepatocellular carcinoma[J].J of Experimental Medicine,1999,189(7):1101-1110.
20.王三英,柳珑,王琳,等.恶性肿瘤患者的抗Koc自身抗体[J].中国免疫学杂志,2002,18:30-32.
21.刘卫红,李志,王鹏,等.肿瘤相关抗原抗体联合检测在肺癌早期诊断中的价值评价[J].中国卫生检验杂志,2006,16(12):1412-1414.
22.Zhang JY,Megliorino R,Peng XX,et al.Antibody detection using tumor-associated antigen mini-array in immunodiagnosing human hepatocelluar carcinoma[J].J Hepatol,2007,46(1):107-114.
23.杜芳,王立东,齐义军,等.食管和贲门癌及癌前病变患者血清中多个自身抗体的检测及临床意义[J].中华肿瘤防治杂志,2006,13(17):1289-1292.
24.Zhang JY,Casiano CA,Peng XX,et al.Enhancement of antibody detection in cancer using panel of recombinant tumor-associated antigens[J].Cancer Epideniol Biomarkers Prev.2003,12(2):136-143.
25.Hunter T,Pines J.Cyclins and cancer Ⅱ:cyclin D and CDK inhibitors come of age[J].Cell,1994,79(4):573-582.
26.董晓,房新志,柴敏秀,等.β-catenin与CyclinD1在乳腺癌中的表达及其意义[J].新疆医科大学学报,2008,31(5):541-547.
27.张海萍,钟山,杨萌霓,等.细胞增殖周期调控因子p16和CyclinD1在食管癌中的表达和意义[J].福建医药杂志,2003,25(3):7-8.
28.Di Raimondo F,Azzaro MP,Palumbo G,et al Angiogenic factors in multiple myeloma:higher levels in bone marrow than in peripheral blood[J].Haematologica.2000.85(8):800-805.
29.郭健政,林茂松,陈卫昌,等.结肠癌组织中p53和CyclinD1基因的表达检测及其临床意义[J].苏州大学学报(医学版),2005,25(5):842-845.
30.郭妍妍.非小细胞肺癌中FHIT、CyclinD1、CDK4的表达及意义[J].中国基层医药,2006,13(12):1994-1995.
31.Kassem SA,Ariel I,Thornton PS,et al.p57(KIP2) expression in normal islet cell and in hyperinsulinism of infancy[J].Diabetes.2001,50(12):2763-2769.
32.Kamura T,Hara T,Kotoshiba S,et al.Degradation of p57kip2 mediated by SCF Skp2-dependent ubiqitylation[J].Proc Natl Acad Sci U S A,2003,100(18):10231-10236.
33.刘勇,路名芝,杨海玉.肺癌中细胞周期调控因子p27和CyclinE的表达及其相关性[J].中国肿瘤临床,2001,28(9):700-701.
34.Blint E,Phillips AC,Kozlov S,et al.Induction of p57(KIP2) expression by p73beta[J].Proc Natl Acad Sci U S A,2002,99(6):3529-3534.
35.Fu MF,Wang CG,Li ZP,et al.CyclinD1:Normal and Abnormal Functions[J].Endocrinology,2004,145(12):5439-5447.
36.Liston P,Roy N,Tamai K,et al Suppression of apoptosis in mammalian cells by NAIP and a related family of IAP genes[J].Nature,1996,379(6563):349-353.
37.Rothe M,Pan MG,Henzel WJ,et al.The TNFR2-TRAF signaling complex contains two novel proteins related to baculoviral inhibitor of apoptosis proteins [J].Cell,1995,83(7):1243-1252.
38.Ambrosini G,Adida C,Altier DC.A novel anti-apoptosis gene,Survivin,expressed in cancer and lymphoma[J].Nat Med,1997,3(8):917-921.
39.Hauser HP,Bardroff M,Pyrowolakis G,et al.A giant ubiquitin-conjugating enzyme related to IAP apoptosis inhibitors[J].J Cell Biol,1998,141(6):1415-1422.
40.Kasof GM,Gomes BC.Livin,a novel inhibitor of.apoptosis protein family member[J].J Biol Chem,2001,276(5):3238-3246.
41.Wang CY,Mayo MW,Korneluk RG,et al.NF-kappaB antiapoptosis:induction of TRAF1 and TRAF2 and c-IAP1 and c-IAP2 to suppress caspase-8 activation[J].Science,1998,281(5383):1680-1683.
42.Rohayem J,Diestelkoetter P,Weigle B,et al.Antibody response to the tumor-associated inhibitor of apoptosis protein Survivin in cancer patients[J].Cancer Res,2000,60(7):1815-1817.
43.王影,梁欢.血清中VEGF、Survivin、p53在肺癌中的表达及相关性[J].中国卫生检验杂志,2008,18(8):1573-1574.
44.Kamb A,Gruis NA,Weaver-Feldhaus J,et al.A cell cycle regulator potentially involved in genesis of many tumor types[J].Science,1994,264(5157):436-440.
45.张军航,马琳,刘芙蓉,等.肺癌组织中p16基因表达的意义[J].中国医科大学学报,2005,34(2):157-158.
46.王海霞,张建勇.FHIT和p16在非小细胞肺癌中的表达及意义[J].山东医药,2008,48(5):37-38.
47.Campbell PM,Der CJ.Oncogenic Ras and its role in tumor cell invasion and metastasis[J].Semin Cancer Biol.2004,14(2):105-114.
48.Tchevkina E,Agapova L,Dyakova N,et al.The small G-protein RalA stimulates metastasis of transformed cells[J].Oncogene,2005,24(3):329-335.
49.Kawana H,Tamaru J,Tanaka T,et al.Role of p27kipl and cyclin-dependent kinase 2 in the proliferation of non-small cell lung cancer[J].Am J Pathol,1998,153(2):505-513.
50.林福地,谢庆祥,张闽峰,等.CyclinE和cdk2在肺癌中表达及其临床意义[J].肿瘤防治杂志,2001,8(3):259-260.
51.Zajac-Kaye M.,Myc oncogene:a key component in cell cycle regulation and its implication for lung cancer[J].Lung Cancer,2001,34(Supp12):S43-46.
1.Jemal A,Tiwaei RC,Murray T,et al.Cancer Statistics,2004[R].CA Cancer J Clin,2004,54(1):8-29.
2.陈娟 综述,张锦 审校.肺癌早期诊断的研究进展[J].肿瘤研究与临床,2003,15(2):134-137.
3.Mountain CF.Revisions in the international system for staging lung cancer[J].Chest,1997,111(6):1710-1717.
4.Martini N,Bains MS,Burt ME,et al.Incidence of local recurrence and second primary tumors in resected stage I lung cancer[J].J Thorac Cardiovasc Surq,1995,109(1):120-9.
5.Winton T,Livingston R,Johnson D,et al.Vinorelbine plus cisplatin vs.observation in resected non-small-cell lung cancer[J].N Eng1 J Med,2005,352(25):2589-2597.
6.Fry WA,Phillips JL,Menck HR,et al.Ten-year survey of lung cancer treatment and survival in hospitals in the United States:a national cancer data base report[J].Cancer,1999,86(9):1867-1876.
7.Gandara DR,Chansky K,Albain KS,et al.Consolidation docetaxel after concurrent chemoradiotherapy in stage IIIB non-small-cell lung cancer:phase ⅡSouthwest Oncology Group Study S9504[J].J Clin Oncol,2003,21(10):2004-2010.
8.Sandler AB,Gray R,Brahmer J,et al.Randomized phase Ⅱ/Ⅲ Trial of paclitaxel (P) plus carboplatin(C) with or without bevacizumab(NSC # 704865) in patients with advanced non-squamous non-small cell lung cancer(NSCLC):An Eastern Cooperative Oncology Group(ECOG) Trial-E4599[R].2005 ASCO Annual Meeting Proceedings.Abstract 4.
9.Tan EM.Autoantibodies in diagnosis and in identifying autoantigens[J].The Immunologist,1999,7:85-92.
10.Tan EM.Autoantibodies in pathology and cell biology[J].Cell,1991,67(5):841-842.
11.Tan EM.Autoantibodies as reporters identifying aberrant cellular mechanisms in tumorigenesis[J].J Clin Invest,2001,108(10):1411-1415.
12.Zhang JY,Chan EK,Peng XX,et al.Autoimmune responses to mRNA binding proteins p62 and Koc in diverse malignancies[J].Clin Immunol,2001,100(2):149-156.
13.Old LJ,Chen YT.New paths in human cancer serology[J].J Exp Med,1998,187(8):1163-1167.
14.陈文彬,潘祥林 主编.诊断书(第六版)[M].北京:人民卫生出版社,2006.
15.Gold P,Freedman SO.Demonstration of tumor specific antigen in human colonic carcinoma by immunologic tolerance and absorption techniques[J].J Exp Med,1965,127:439-462.
16.Ando S,Kimura H,Iwai N,et al.Optimal combination of seven tumor markers in prediction of advanced stage at first examination of patients with no-small cell lung cancer[J].Anticancer Res,2001,21(4B):3085-3092.
17.Kulpa J,Wojcik E,Reinfuss M,et al.Carcinoembryonic antigen,squamous cell carcinoma antigen,CYFRA21-1,and neuron-specific enolase in squamous cell ling cancer patients[J].Clin Chem,2002,48(11):1931-1937.
18.Mitsuhashi N,Takahashi T,Sakurai H,et al.Establishment and characterization of a new human lung poorly differentiated adenocarcinoma cell line,GLL-1,producing carcinoem-bryonic antigen(CEA) and CA19-9[J].Lung cancer,1995,12(1-2):13-24.
19.卞宝祥,胡楠,吴风雷,等.血清和胸腔积液CEA、NSE、CYFRA21-1、SCC-Ag联检对肺癌诊断的临床意义[J].放射免疫学杂志,2008,21(5):391-394.
20.Kato H,Morioka H,Aramaki S,et al.Radioimmunoassay for tumor antigen of human cervical squamous cell carcinoma[J].Cell Mol Biol Incl Cyto Enzymol,1979,25(1):51-56.
21.李斌,周海中,曹剑霞.SCC-Ag、CA19-9、CEA在肺癌患者血清中的变化及临床意义[J].放射免疫学杂志,2007,20(5):464-466.
22.刘举珍,蔡铁铁,秦莎娜.血清NSE、CA242、TPA、CEA联检对肺癌的诊断价值[J].放射免疫学杂志,2007,20(4):371-372.
23.黄文焰,郑军.血清肿瘤标志物检测对肺癌早期诊断的意义[J].临床肺科杂志,2008,13(12):1620-1621.
24.韩丹,陈志飞,邱鸣.CA-125在肺结核、结核性胸膜炎与肺癌鉴别诊断中的价值[J].临床肺科杂志,2008,13(9):1160-1161.
25.Alatas F,Alatas O,Metintas M.Diagnostic value of CEA,CA15-3,CA19-9,CYFRA21-1,NSE and TSA assay inpleural effusions[J].Lung cancer,2001,31(1):9-16.
26.谭丽珍,冯献斌,范先基,等.非小细胞肺癌患者血清CEA、CA-199的临床意义[J].广西医学,2009,31(1):59-60.
27.Miedouge M,Devys A,Simon M,et al.High levels of cytokeratin 19 fragments but no evidence of cytokeratins 1,2,10/11,14 or filaggrin in the serum of squamous cell lung carcinoma patients[J].Tumor Biol,2001,22(1):19-26.
28.李世生,黄婵娟.联合检测血清CYFRA21-1、CA125和NSE对肺癌诊断的价值[J].现代肿瘤医学,2008,16(6):951-953.
29.Plebani M,Basso D,Navaqlia F,et al.Clinical evalution of seven tumor markers in lung cancer diagnosis can any combination improve the results?[J].Br J Cancer,1995,72(1):170-173.
30.宋鉴清,赵敏,庞鹿.血清CYFRA21-1、NSE检测在肺癌诊断中的应用[J].实用肿瘤学杂志,2003,17(2):151-152.
31.Schneider J,Velcovsky HG,Morr H,et al.Comparison of the tumor markers tumor M2-PK,CEA,CYFRA21-1,NSE and SCC in the diagnosis of lung cancer [J].Anticancer Res,2000,20(6D):5053-5058.
32.黄文成,陈闯,朱波,等.ECLIA法测定肺癌患者血清NSE和CYFRA21-1的临床意义[J].现代肿瘤医学,2008,16(1):41-44.
33.Mitza A,McGuirk M,Hockenberry TN,et al.Human Surviving is negatively regulated by wild-type p53 and participates in p53 dependent apoptotic pathway [J].Oncogene,2002,21(17):2613-2622.
34.Zajac-Kaye M.Myc oncogene:a key component in cell cycle regulation and its implication for lung cancer[J].Lung Cancer,2001,34(Suppl 2):S43-46.
35.Graziano SL,Gamble GP,Newman NB,et al.Prognostic significance of K-ras codon 12 mutations in patients with resected stage Ⅰ and Ⅱ non-small-cell lung cancer[J].J Clin Oncol,1999,17(2):668-675.
36.Charoenkwan P,Senger C,Weitnzma S,et al.Significance of p53 expression in immature teratomas[J].Pediatr Dev Pathol,2002,5(5):499-507.
37.王影,梁欢.血清中VEGF、Survivin、p53在肺癌中的表达及相关性[J].中国卫生检验杂志,2008,18(8):1573-1574.
38.Kamb A,Gruis NA,Weaver-Feldhaus J,et al.A cell cycle regulator potentially involved in genesis of many tumor types[J].Science,1994,264(5157):436-440.
39.Bearzatto A,Conte D,Frattini M,et al P16(INK4A) Hypemethylation detected by fluorescent methylation-specific PCR in plasmas from non-small cell lung cancer[J].Clin Cancer Res,2002,8(12):3782-3787.
40.张军航,马琳,刘芙蓉,等.肺癌组织中p16基因表达的意义[J].中国医科大学学报,2005,34(2):157-158.
41.王海霞,张建勇.FHIT和p16在非小细胞肺癌中的表达及意义[J].山东医药,2008,48(5):37-38.
42.Hunter T,Pines J.Cyclins and cancer Ⅱ:cyclinD and CDK inhibitors come of age[J].Cell,1994,79(4):573-582.
43.董晓,房新志,柴敏秀,等.β-catenin与CyclinD1在乳腺癌中的表达及其意义[J].新疆医科大学学报,2008,31(5):541-547.
44.张海萍,钟山,杨萌霓,等.细胞增殖周期调控因子p16和CyclinD1在食管癌中的表达和意义[J].福建医药杂志,2003,25(3):7-8.
45.Di Raimondo F,Azzaro MP,Palumbo G;et al Angiogenic factors in multiple myeloma:higher levels in bone marrow than in peripheral blood[J].Haematologica,2000,85(8):800-805.
46.郭健政,林茂松,陈卫昌,等.结肠癌组织中p53和CyclinD1基因的表达检测及其临床意义[J].苏州大学学报(医学版),2005,25(5):842-845.
47.郭妍妍.非小细胞肺癌中FHIT、CyclinD1、CDK4的表达及意义[J].中国基层医药,2006,13(12):1994-1995.
48.Fu MF,Wang CG,Li ZP,et al.CyclinD1:Normal and Abnormal Functions[J].Endocrinology,2004,145(12):5439-5447.
49.王鸿程,许学亮,简丽萍,等.STAT5与CyclinB1、CyclinD1、c-los、C-myc 在非小细胞肺癌中的表达及临床意义[J].泸州医学院学报,2008,31(6):601-605.
50.杜芳,王立东,齐义军,等.食管和贲门癌及癌前病变患者血清中多个自身抗体的检测及临床意义[J].中华肿瘤防治杂志,2006,13(17):1289-1292.
51.刘卫红,李志,王鹏,等.肿瘤相关抗原抗体联合检测在肺癌早期诊断中的价值评价[J].中国卫生检验杂志,2006,16(12):1412-1414.
52.Kassem SA,Ariel I,Thornton PS,et al.p57(KIP2) expression in normal islet cell and in hyperinsulinism of infancy[J].Diabetes.2001,50(12):2763-2769.
53.Kamura T,Hara T,Kotoshiba S,et al.Degradation of p57kip2 mediated by SCF Skp2-dependent ubiqitylation[J].Proc Natl Acad Sei,2003,100(18):10231-10236.
54.刘勇,路名芝,杨海玉.肺癌中细胞周期调控因子p27和CyclinE的表达及其相关性[J].中国肿瘤临床,2001,28(9):700-701.
55.Blint E,Phillips AC,Kozlov S,et al.Induction of p57(KIP2) expression by p73beta[J].Proc Natl Acad Sci,2002,99(6):3529-3534.
56.Kawana H,Tamaru J,Tanaka T,et al.Role of p27kipl and cyclin-dependent kinase 2 in the proliferation of non-small cell lung cancer[J].Am J Pathol 1998,153(2):505-513.
57.林福地,谢庆祥,张闽峰,等.CyclinE和cdk2在肺癌中表达及其临床意义[J].肿瘤防治杂志,2001,8(3):259-260.
58.Rohayem J,Diestelkoetter P,Weigle B,et al.Antibody response to the tumor-associated inhibitor of apoptosis protein Survivin in cancer patients[J].Cancer Res,2000,60(7):1815-1817.
59.Akyurek N,Memis L,Ekinci O,et al.Survivin expression in pre-invasive lesions and non-small cell lung carcinoma[J].Virchows Arch,2006,449(2):164-170.
60.Kim NW,Piatyszek MA,Prouse KR,et al.Specific association of human telomerase activity with immortal cells and cancer[J].Science 1994,266(5193):2011-2015.
61.Albanell J,Lonardo F,Rusch V,et al.High Telomerase activity in primary lung cancer:association with increased cell proliferation rates and advanced pathologic state[J].J Natl Cancer Inst,1997,89(21):1609-1615.
62.刘瑾,杨青,兰亚佳,等.人体肺癌端粒酶表达的研究[J].现代预防医学,2004,31(3):317-319.