肝癌相关抗原KINECTIN的D2与D3选择剪接区在人肝癌与正常肝组织表达差异性的研究
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摘要
目的:检测人类动力素蛋白(kinectin)D2与D3选择剪接区(alternative splicing site)在人肝细胞癌(HCC)组织与癌旁组织以及正常肝组织中表达的差异性,探索kinectin的选择剪接区与肝癌发生之间可能存在的联系。
方法:用RT-PCR法获取45例肝癌组织标本和癌旁组织标本以及10例正常肝组织标本的cDNA,并用半定量PCR法分别检测其D2选择剪接区(以下简称D2区)和D3选择剪接区(以下简称D3区)表达情况的差异性;将MBP-kinectin融合蛋白作为抗原,通过ELISA技术检测45例肝癌患者血清标本中kinectin抗体的阳性率,并进行统计学分析。
结果:肝癌组织D2~L/D2~S比率高于癌旁组织和正常肝组织,差异有统计学意义;D3~L/D3~S比率在肝癌组织、癌旁组织及正常肝组织之间无明显差异;肝癌组织D2~L/D2~S比率与血清中kinectin抗体滴度值无明显相关性。
结论:包含D2区的kinectin异型体在肝癌组织中表达水平增高,提示其可能与肝癌的发生有一定的相关性。
Objective: Analyze the difference in expression of D2 and D3 alternative splicing sites of human kinectin in human hepatocellular carcinoma(HCC)tissues, adjacent non-cancerous tissues and normal liver tissues,and investigate the possible relationship between alternative splicing sites of kinectin and the hepatocarcinogenesis.
Methods: The cDNA was obtained by RT-PCR in 45 coupled HCC cancerous and adjacent tissues,and 10 normal liver tissues.The difference in the expression of D2 and D3 alternative splicing sites in cDNA was examined by semi-quantitative PCR,fusion protein MBP-kinectin which was obtained by bio-engineering was used as the antigen. The positive rate of anti-kinectin in hepatocellular cancerous serum of 45 patients were detected by ELISA.Then,statistical analysis was performed.
Results: The ratio of D2~L/D2~S in hepatocellular cancerous tissues is higher than that of adjacent non-cancerous tissues and normal liver tissues.There isstatistical significance between them.The ratio of D3~L /D3~S has no Significant difference among hepatocellular cancerous tissues ,adjacent non-cancerous tissues and normal liver tissues.There is no obvious correlation between the ratio of D2~L/D2~S in hepatocellular cancerous tissues and the titers of anti-kinectin in coupled serum by ELISA.
Conclusion: Variant containing D2 is overexpressed in cancerous tissues and this alteration may be tumor associated.
方法:用RT-PCR法获取45例肝癌组织标本和癌旁组织标本以及10例正常肝组织标本的cDNA,并用半定量PCR法分别检测其D2选择剪接区(以下简称D2区)和D3选择剪接区(以下简称D3区)表达情况的差异性;将MBP-kinectin融合蛋白作为抗原,通过ELISA技术检测45例肝癌患者血清标本中kinectin抗体的阳性率,并进行统计学分析。
结果:肝癌组织D2~L/D2~S比率高于癌旁组织和正常肝组织,差异有统计学意义;D3~L/D3~S比率在肝癌组织、癌旁组织及正常肝组织之间无明显差异;肝癌组织D2~L/D2~S比率与血清中kinectin抗体滴度值无明显相关性。
结论:包含D2区的kinectin异型体在肝癌组织中表达水平增高,提示其可能与肝癌的发生有一定的相关性。
Objective: Analyze the difference in expression of D2 and D3 alternative splicing sites of human kinectin in human hepatocellular carcinoma(HCC)tissues, adjacent non-cancerous tissues and normal liver tissues,and investigate the possible relationship between alternative splicing sites of kinectin and the hepatocarcinogenesis.
Methods: The cDNA was obtained by RT-PCR in 45 coupled HCC cancerous and adjacent tissues,and 10 normal liver tissues.The difference in the expression of D2 and D3 alternative splicing sites in cDNA was examined by semi-quantitative PCR,fusion protein MBP-kinectin which was obtained by bio-engineering was used as the antigen. The positive rate of anti-kinectin in hepatocellular cancerous serum of 45 patients were detected by ELISA.Then,statistical analysis was performed.
Results: The ratio of D2~L/D2~S in hepatocellular cancerous tissues is higher than that of adjacent non-cancerous tissues and normal liver tissues.There isstatistical significance between them.The ratio of D3~L /D3~S has no Significant difference among hepatocellular cancerous tissues ,adjacent non-cancerous tissues and normal liver tissues.There is no obvious correlation between the ratio of D2~L/D2~S in hepatocellular cancerous tissues and the titers of anti-kinectin in coupled serum by ELISA.
Conclusion: Variant containing D2 is overexpressed in cancerous tissues and this alteration may be tumor associated.
引文
1. Wang HC, Su YR, Han KJ, Pang XW, Peng JR, Liang B, Wang S, Chen WF. Multiple variants and a differential splicing pattern of kinectin in human hepatocellular carcinoma. Biochem Cell Biol. 2004 Apr;82(2):321-327.
2. Lawrence S.B. Goldstein , Shermali Gunawardena, et al. Flying Through the Drosophila Cytoskeletal Genome. The Journal of Cell Biology, 2000,150 (2): 63-68.
3. A Fiitterer, G Kruppa, B Kramer, H Lemke, and M Kronke. Molecular cloning and characterization of human kinectin. Mol Biol Cell. 1995 February; 6(2): 161-170.
4. Vignal, A. Blangy,M. Martin, C. Gauthier-Rouviere, and P. Fort Kinectin Is a Key Effector of RhoG Microtubule-Dependent Cellular Activity Molecular and Cellular Biology, December 2001, Vol. 21, No. 23,8022-8034,
5. Huan Tran, Roumen Pankov, Simon D. Tran, Brian Hampton, Wilson H. Burgess and Kenneth M. Yamada, Integrin clustering induces kinectin accumulation. Journal of Cell Science 115,2031-2040 (2002).
6. Lee-Lee Ong, Connie P. N. Er, Andrea Ho. May T. Aung and Hanry Yu .Kinectin Anchors the Translation Elongation Factor-1d to the Endoplasmic Reticulum J. Biol. Chem., Vol. 278, Issue 34, 32115-32123, August 22, 2003.
7. Niovi Santama, Connie P. N. Er, Lee-Lee Ong and Hanry Yu Distribution and functions of kinectin isoforms Journal of Cell Science 117, 4537-4549 (2004).
8. Fan Yang, Irene H. Chen, Zeyu Xiong etc. Model of stimulation-responsive splicing and strategies in identification of immunogenic isoforms of tumor antigens and autoantigens Clinical Immunology Volume 121,Issue 2,November 2006,Pages 121-133.
9.Barmak Modrek,and Christopher Lee.A genomic view of alternative splicing.Nature Genetics,2002,30:13-19.
10.章国卫,宋怀东,陈竺.选择性剪接的分子机制[J].遗传学报,2004,31(1):102-107
11.Gravley BR.Alternative splicing:increasing diversity in the proteomic world.Trends Genet,2001,17:100-107.
12.Lain BJ,Hertel KJ.A general role for splicing enhancers in exon deftnition.RNA,2002,8:1233-1241.
13.Li B,Wachtel C,Miriami E,et al.Stop codons affect 5'splice site selection by surveillance of splicing.Proc Natl Acad Sci USA,2002,99(8):5277-5282.
14.Goldstrohm AC,Greenleaf AL,Garcia-Blanco MA.Co-transcriptioal splicing of pre.messenger RNAs:considerations for the mechanism of alternative splicing.Gene,2001,277:31-47.
15.YANG Jin-Chun,CHEN Yu-Hua,MO Jin-Dan,CHEN Cheng The research progress on alternative splicing of pre-mRNA Life Science,2002,4(3):150-153.
16.Zhou Z,Licklider LJ,Gygi SP,et al.Comprehensive proteomic analysis of the human spliceosome.Nature,2002,419:182-185.
17.Makarova OV,Makarov EM,Liu S,et al.Protein 61K,encoded by a gene(PRPF31)linked to autosomal dominant retinitis pigmentosa,is required for U4/U6~*U5 tri-snRNP formation and pre-mRNA splicing.EMBO J,2002,21:1148-1157.
18.Dasen JS,O'Connel SM,Flyrm SE,et al.Reciprocal interactions of Pitl and GATA2 mediate signaling gradient.induced determination of pituitary cell types.Cel,1999,28;97(5):587-598.
19.Sigalas I,Calvert AH,Anderson JJ,et al.Alternatively spliced mdm2 transcripts with loss of p53 binding domain sequences: transforming ability and frequent detection in human cancer.Nat Med,1996,2:912-917.
20.ebhardt F,Zanker KS,Brandt B.Differential expression of alternatively spliced c-erbB-2 mRNA in primary tumors,lymph node metastases, and hone marrow micrometastases from breast cancer patient.Biochem Biophys Res Commun,1998,247:319-323.
21.Heller S, Scheibenpflug L,Westermark B,et al.PDGF B mRNA variants in human tumors with similarity to the v-sis oncogene: expression of cellular PDGF B protein is associated with exon 1 divergence, but not with a 3'UTR splice variant,Int J Cancer,2000,85:211-222.
22.Narla G,Difeo A,Reeves HL,et al.A germline DNA polymorphism enhances alternative splicing of the KLF6 tumor suppressor gene and is associated with increased prostate cancer risk.Cancer Res,2005,65:1213-1222.
23.Baudry D,Faussillon M,Cabanis MO,et al.Changes in WTI splicing are associated with a specific gene expression profile in Wilms' tumour. Oncogene,2002,21:5566-5573.
24.Xin Y, Grace A,Gallagher MM,et al.CD44v6 in gastric carcinoma:a marker of tumor progression.Appl Immunohistochem Mol Morphol,2001,9:138-142.
25.Wang L,Duke L,Zhang PS,et al.Alternative splicing disrupts a nuclear localization signal in spleen tyrosine kinase that is required for invasion suppression in breast cancer.Cancer Res,2003,63:4724-4730.
1.Sahin U,Tureci o,Schmitt H,et al.Human neoplasms elicit multiphe specific immune responses in the autologous host[J].Proc Natl Acad Sci USA,1995,92(25):11810-11813.
2.Scanlan MJ,Gordan JD,Williamson B,et al.Antigens recognized by autologous antibody in patients with renal-cell carcinoma[J].Int J Cancer,1999,83(4):456-464.
3.Jager D,Stockert E,Gure AO,et al.Identification of a tissue-specific putative transcription factor in breast tissue by serological screening of a breast cancer library[J].Cancer Res,2001,61:2055-2061.
4.Chen YT,Scanlan M J,Obata Y,et al.Identification of human tumor antigens by serological expression cloning.Principles and Practice of Biologic Therapy of Cancer.Rosenberg SA.Philadelphia PA[J].Lippincott Williams Wilkins.2000,557-570.
5.Scanlan MJ,Chen YT,Williamson B,et al.Characterization of human colon cancer antigens recognized by autologous antibodies.Int J Cancer,1998,76:652-658.
6.Obata Y,Takahashi T,Sakamoto J,et al.SEREX analysis of gastric cancer antigens[J].Cancer Chemother Pharmacol,2000,46(suppl):37-42.
7.Tetsuya N,Satoru S,Kazuhiro Y,et al.Gene cloning of immunogenic antigens overexpressed in pancreatic cancer[J].Biochen Biophys Res Commun,2001,281(4):936-944.
8.罗国容,刘启福,Stenner F,等.自体血清抗体筛选人肝细胞癌抗原的分子辩认[J].中华医学杂志,1999,79(6):447.
9.王欲,韩克军,屈伟,陈慰峰.新型肝癌相关性抗原HCA519及其变异体 HCA90基因克隆和表达.中国生物化学与分子生物学报,2003,Vol.19No.1 P.46-52.
10.邢宝才,袁艳华,胡志国,刘月香,韩燕,刘年,陈慰峰,王歈.NY-ESO-1/LAGE-1基因在肝癌组织中的表达及其诱导的体液免疫应答.实用癌症杂志,2004,Vol.84 No.23 P.1980-1982.
11.Bing Li,Xiao-Ping Qian,Xue-Wen Pang,Wan-Zhong Zou,Yu-Ping Wang,Hong-Yan Wu,and Wei-Feng Chen HCA587 Antigen Expression in Normal Tissues and Cancers:Correlation with Tumor Differentiation in Hepatocellular Carcinoma LABORATORY INVESTIGATION 2003Vol.83 No.8 P.1185-1191.
12.王宏程,杨慕华,石永玉,庞学雯,杨小昂,彭吉润,冷希圣,陈慰峰.肝细胞癌相关肿瘤抗原基因的筛选与分析.中华肝脏病杂志,2005,Vol.13 No.5P.343-346.
13.王颖,葛海良,袁明,张惠珍,王树军.从卵巢癌cDNA文库中筛选新的肿瘤标志物.上海免疫学杂志,2002,Vol.22 No.6 P.371-374.
14.周芙玲,张王刚,陈刚,赵万红,曹星梅,陈银霞,田纬,刘苏虎,吴明霞,刘明.多发性骨髓瘤相关抗原的免疫学筛选及生物信息学分析.中国实验血液学杂志,2006,Vol.14 No.2:P.252-257.
15.谢可,田聆,魏于全,邓洪新,赵霞,李炯,文艳君.鼻咽癌相关抗原的血清学筛选与鉴定.实用医院临床杂志,2004,Vol.1 No.2 P.27-29.
16.黄绍明,何少健,莫发荣,梁皓,廖红,谢小薰,罗国容.应用SEREX技术筛选与鉴定鼻咽癌相关抗原.广西医科大学学报,2005,Vol.22 No.2P.166-169.
17.舒峻,李官成,何小鹃.鼻咽癌组织cDNA文库的构建及抗原基因的筛选.生物工程学报,2004,Vol.20 No.1 P.371-374.
18.何敬东,周高速,肖冰,李旭,刘宇虎,张振书,南清振.从大肠癌组织cDNA 表达文库筛选肿瘤抗原基因.解放军医学杂志,2005,Vol.30 No.5P.382-384.
19.Scanlan MJ,Chen YT,Williamson B,et al.Characterization of human colon canncer antigens recognized by autologous antibodies[J].Int J Cancer,1998,76(5):652-658.
20.Gure AO,Altorki NK,Stoekert E,et al.Human lung cancer antigens recognized by autologous antibodies:definition of a novel cDNA derived from the tumor suppressor gene locus on chromosome 3p21.3[J].Cancer Res,1998,58(5):1034-1041.
21.Amy L,Danid D,Dipetrillo T,et al.Emergence of increased cerebral metastasis after high-dose preoperative radiotherapy with chemotherapy in patients with locally advavced non-small cell lung carcinoam[J].Cancer,2001,92(1):160-164.
22.Itoh M,Watanabe M,Yamada Y,et al.HUB1 is an autoantigen frequently eliciting humoral immune response in patients with adult T cell leukemia [J].Int J Oncol,1999,14(4):703-708.
23.王运伟,朱正纲,顾琴龙,李建芳,陈雪华,刘炳亚,林言箴.SEREX方法鉴定胃癌肿瘤相关抗原基因.上海第二医科大学学报,2005,Vol.25 No.3P.235-238.
2. Lawrence S.B. Goldstein , Shermali Gunawardena, et al. Flying Through the Drosophila Cytoskeletal Genome. The Journal of Cell Biology, 2000,150 (2): 63-68.
3. A Fiitterer, G Kruppa, B Kramer, H Lemke, and M Kronke. Molecular cloning and characterization of human kinectin. Mol Biol Cell. 1995 February; 6(2): 161-170.
4. Vignal, A. Blangy,M. Martin, C. Gauthier-Rouviere, and P. Fort Kinectin Is a Key Effector of RhoG Microtubule-Dependent Cellular Activity Molecular and Cellular Biology, December 2001, Vol. 21, No. 23,8022-8034,
5. Huan Tran, Roumen Pankov, Simon D. Tran, Brian Hampton, Wilson H. Burgess and Kenneth M. Yamada, Integrin clustering induces kinectin accumulation. Journal of Cell Science 115,2031-2040 (2002).
6. Lee-Lee Ong, Connie P. N. Er, Andrea Ho. May T. Aung and Hanry Yu .Kinectin Anchors the Translation Elongation Factor-1d to the Endoplasmic Reticulum J. Biol. Chem., Vol. 278, Issue 34, 32115-32123, August 22, 2003.
7. Niovi Santama, Connie P. N. Er, Lee-Lee Ong and Hanry Yu Distribution and functions of kinectin isoforms Journal of Cell Science 117, 4537-4549 (2004).
8. Fan Yang, Irene H. Chen, Zeyu Xiong etc. Model of stimulation-responsive splicing and strategies in identification of immunogenic isoforms of tumor antigens and autoantigens Clinical Immunology Volume 121,Issue 2,November 2006,Pages 121-133.
9.Barmak Modrek,and Christopher Lee.A genomic view of alternative splicing.Nature Genetics,2002,30:13-19.
10.章国卫,宋怀东,陈竺.选择性剪接的分子机制[J].遗传学报,2004,31(1):102-107
11.Gravley BR.Alternative splicing:increasing diversity in the proteomic world.Trends Genet,2001,17:100-107.
12.Lain BJ,Hertel KJ.A general role for splicing enhancers in exon deftnition.RNA,2002,8:1233-1241.
13.Li B,Wachtel C,Miriami E,et al.Stop codons affect 5'splice site selection by surveillance of splicing.Proc Natl Acad Sci USA,2002,99(8):5277-5282.
14.Goldstrohm AC,Greenleaf AL,Garcia-Blanco MA.Co-transcriptioal splicing of pre.messenger RNAs:considerations for the mechanism of alternative splicing.Gene,2001,277:31-47.
15.YANG Jin-Chun,CHEN Yu-Hua,MO Jin-Dan,CHEN Cheng The research progress on alternative splicing of pre-mRNA Life Science,2002,4(3):150-153.
16.Zhou Z,Licklider LJ,Gygi SP,et al.Comprehensive proteomic analysis of the human spliceosome.Nature,2002,419:182-185.
17.Makarova OV,Makarov EM,Liu S,et al.Protein 61K,encoded by a gene(PRPF31)linked to autosomal dominant retinitis pigmentosa,is required for U4/U6~*U5 tri-snRNP formation and pre-mRNA splicing.EMBO J,2002,21:1148-1157.
18.Dasen JS,O'Connel SM,Flyrm SE,et al.Reciprocal interactions of Pitl and GATA2 mediate signaling gradient.induced determination of pituitary cell types.Cel,1999,28;97(5):587-598.
19.Sigalas I,Calvert AH,Anderson JJ,et al.Alternatively spliced mdm2 transcripts with loss of p53 binding domain sequences: transforming ability and frequent detection in human cancer.Nat Med,1996,2:912-917.
20.ebhardt F,Zanker KS,Brandt B.Differential expression of alternatively spliced c-erbB-2 mRNA in primary tumors,lymph node metastases, and hone marrow micrometastases from breast cancer patient.Biochem Biophys Res Commun,1998,247:319-323.
21.Heller S, Scheibenpflug L,Westermark B,et al.PDGF B mRNA variants in human tumors with similarity to the v-sis oncogene: expression of cellular PDGF B protein is associated with exon 1 divergence, but not with a 3'UTR splice variant,Int J Cancer,2000,85:211-222.
22.Narla G,Difeo A,Reeves HL,et al.A germline DNA polymorphism enhances alternative splicing of the KLF6 tumor suppressor gene and is associated with increased prostate cancer risk.Cancer Res,2005,65:1213-1222.
23.Baudry D,Faussillon M,Cabanis MO,et al.Changes in WTI splicing are associated with a specific gene expression profile in Wilms' tumour. Oncogene,2002,21:5566-5573.
24.Xin Y, Grace A,Gallagher MM,et al.CD44v6 in gastric carcinoma:a marker of tumor progression.Appl Immunohistochem Mol Morphol,2001,9:138-142.
25.Wang L,Duke L,Zhang PS,et al.Alternative splicing disrupts a nuclear localization signal in spleen tyrosine kinase that is required for invasion suppression in breast cancer.Cancer Res,2003,63:4724-4730.
1.Sahin U,Tureci o,Schmitt H,et al.Human neoplasms elicit multiphe specific immune responses in the autologous host[J].Proc Natl Acad Sci USA,1995,92(25):11810-11813.
2.Scanlan MJ,Gordan JD,Williamson B,et al.Antigens recognized by autologous antibody in patients with renal-cell carcinoma[J].Int J Cancer,1999,83(4):456-464.
3.Jager D,Stockert E,Gure AO,et al.Identification of a tissue-specific putative transcription factor in breast tissue by serological screening of a breast cancer library[J].Cancer Res,2001,61:2055-2061.
4.Chen YT,Scanlan M J,Obata Y,et al.Identification of human tumor antigens by serological expression cloning.Principles and Practice of Biologic Therapy of Cancer.Rosenberg SA.Philadelphia PA[J].Lippincott Williams Wilkins.2000,557-570.
5.Scanlan MJ,Chen YT,Williamson B,et al.Characterization of human colon cancer antigens recognized by autologous antibodies.Int J Cancer,1998,76:652-658.
6.Obata Y,Takahashi T,Sakamoto J,et al.SEREX analysis of gastric cancer antigens[J].Cancer Chemother Pharmacol,2000,46(suppl):37-42.
7.Tetsuya N,Satoru S,Kazuhiro Y,et al.Gene cloning of immunogenic antigens overexpressed in pancreatic cancer[J].Biochen Biophys Res Commun,2001,281(4):936-944.
8.罗国容,刘启福,Stenner F,等.自体血清抗体筛选人肝细胞癌抗原的分子辩认[J].中华医学杂志,1999,79(6):447.
9.王欲,韩克军,屈伟,陈慰峰.新型肝癌相关性抗原HCA519及其变异体 HCA90基因克隆和表达.中国生物化学与分子生物学报,2003,Vol.19No.1 P.46-52.
10.邢宝才,袁艳华,胡志国,刘月香,韩燕,刘年,陈慰峰,王歈.NY-ESO-1/LAGE-1基因在肝癌组织中的表达及其诱导的体液免疫应答.实用癌症杂志,2004,Vol.84 No.23 P.1980-1982.
11.Bing Li,Xiao-Ping Qian,Xue-Wen Pang,Wan-Zhong Zou,Yu-Ping Wang,Hong-Yan Wu,and Wei-Feng Chen HCA587 Antigen Expression in Normal Tissues and Cancers:Correlation with Tumor Differentiation in Hepatocellular Carcinoma LABORATORY INVESTIGATION 2003Vol.83 No.8 P.1185-1191.
12.王宏程,杨慕华,石永玉,庞学雯,杨小昂,彭吉润,冷希圣,陈慰峰.肝细胞癌相关肿瘤抗原基因的筛选与分析.中华肝脏病杂志,2005,Vol.13 No.5P.343-346.
13.王颖,葛海良,袁明,张惠珍,王树军.从卵巢癌cDNA文库中筛选新的肿瘤标志物.上海免疫学杂志,2002,Vol.22 No.6 P.371-374.
14.周芙玲,张王刚,陈刚,赵万红,曹星梅,陈银霞,田纬,刘苏虎,吴明霞,刘明.多发性骨髓瘤相关抗原的免疫学筛选及生物信息学分析.中国实验血液学杂志,2006,Vol.14 No.2:P.252-257.
15.谢可,田聆,魏于全,邓洪新,赵霞,李炯,文艳君.鼻咽癌相关抗原的血清学筛选与鉴定.实用医院临床杂志,2004,Vol.1 No.2 P.27-29.
16.黄绍明,何少健,莫发荣,梁皓,廖红,谢小薰,罗国容.应用SEREX技术筛选与鉴定鼻咽癌相关抗原.广西医科大学学报,2005,Vol.22 No.2P.166-169.
17.舒峻,李官成,何小鹃.鼻咽癌组织cDNA文库的构建及抗原基因的筛选.生物工程学报,2004,Vol.20 No.1 P.371-374.
18.何敬东,周高速,肖冰,李旭,刘宇虎,张振书,南清振.从大肠癌组织cDNA 表达文库筛选肿瘤抗原基因.解放军医学杂志,2005,Vol.30 No.5P.382-384.
19.Scanlan MJ,Chen YT,Williamson B,et al.Characterization of human colon canncer antigens recognized by autologous antibodies[J].Int J Cancer,1998,76(5):652-658.
20.Gure AO,Altorki NK,Stoekert E,et al.Human lung cancer antigens recognized by autologous antibodies:definition of a novel cDNA derived from the tumor suppressor gene locus on chromosome 3p21.3[J].Cancer Res,1998,58(5):1034-1041.
21.Amy L,Danid D,Dipetrillo T,et al.Emergence of increased cerebral metastasis after high-dose preoperative radiotherapy with chemotherapy in patients with locally advavced non-small cell lung carcinoam[J].Cancer,2001,92(1):160-164.
22.Itoh M,Watanabe M,Yamada Y,et al.HUB1 is an autoantigen frequently eliciting humoral immune response in patients with adult T cell leukemia [J].Int J Oncol,1999,14(4):703-708.
23.王运伟,朱正纲,顾琴龙,李建芳,陈雪华,刘炳亚,林言箴.SEREX方法鉴定胃癌肿瘤相关抗原基因.上海第二医科大学学报,2005,Vol.25 No.3P.235-238.