荧光原位杂交结合细胞形态学分析食管癌脱落细胞染色体改变的研究
详细信息    本馆镜像全文|  推荐本文 |  |   获取CNKI官网全文
摘要
背景与目的:食管癌细胞学诊断中常见到低度鳞状上皮内病变(LSIL)、非典型鳞状上皮细胞(ASC)或形态改变更轻微的细胞,这类细胞是否发生了癌变相关的分子改变尚不清楚。本研究旨在利用细胞形态学相结合的荧光原位杂交(FISH)技术检测LSIL/ASC细胞的染色体改变。
     方法:选择食管鳞癌手术切除组织癌肿表面刮取细胞标本67例及其癌旁粘膜表面细胞标本30例,经液基制片,进行巴氏染色后,采用8、10和11号染色体着丝粒探针,进行FISH检测,分析检测结果与临床病理参数之间的关系。
     结果:在癌肿表面细胞标本,癌细胞/可疑癌细胞(Ca.C/Suca.C)8、10和11号染色体表现非整倍体病例阳性率分别为98.4%、83.3%和96.1%,LSIL/ASC各为68.8%、55.6%和64.3%,Ca.C/Suca.C显著高于LSIL/ASC(P<0.05)。在癌旁细胞标本,LSIL/ASC细胞中三条染色体非整倍体的阳性病例率分别为66.7%、46.2%和53.3%。癌肿表面与癌旁LSIL/ASC染色体异常无统计学差异(P>0.05)。癌肿表面LSIL/ASC细胞中8、10号染色体同时畸变病例率与局部淋巴结转移及高临床分期正相关。所有标本中正常细胞均未见染色体畸变
     结论:食管脱落细胞液基薄片上可以建立与细胞形态相结合的FISH技术。食管鳞癌表面脱落的癌细胞及LSIL/ASC均可能发生8、10和11号染色体非整倍体改变,与细胞形态相结合的FISH技术可能辅助食管癌细胞学早期诊断。
Objective:Low grade squamous intraepithelial lesion(LSIL),atypical squamous cell(ASC) or cells with slightly changes in morphology were often observed in cytological diagnosis.It remains unclear whether there occurred molecular changes related with carcinogenesis in these kinds of cells.In the present study,we explore the feasibility of fluorescence in situ hybridization(FISH) combined with cyto-morphology in the detection of chromosome aberrations in exfoliated cells from esophageal squamous carcinoma(ESCC).
     Methods:The specimens of liquid-based cytology were prepared by scraping from the surface of 67 tumor and 30 adjacent mucosa.After Pap staining,chromsome aberrations were detected with centromeric probes specific for chromosmes 8,10 and 11 by FISH.The correlations between these data and the clinicopathologic parameters were analysed.
     Results:The aneuploidy of chromosmes 8,10 and 11 were observed in cancerous and suspencious cancerous cells(Ca.C/Suca.C) of 98.4%,83.3%and 96.1%cases,and in LSIL/ASC of 68.8%,55.6%and 64.3%cases,respectively.The LSIL/ASC of 66.7%, 46.2%and 53.3%adjacent mucosa presented abnormality of the three chromosmes. There were siganificant higher aneuploidy rates in Ca.C/Suca.C than in LSIL/ASC(P<0.05),but no statistic difference between LSIL/ASC exfoliated from the surface of tumor and from the adjacent mucosa.The simultaneous aberrations of chromosmes 8 and 10 were detected predominantly in LSIL/ASC from the surface of tumor with local lymph node metastasis and later clinical stages.No chromosome aberrations were found in all normal squmous epithelium.
     Conclusions:FISH technique combined with cytologic morphology could be established successfully in the specimens of liquid-based cytologic.The aberrations involving chromosomes 8,10 and 11 occurred in Ca.C/Suca.C and LSIL/ASC exfoliated from the suface of ESCC tissues.FISH combined with cytologic morphology could adjuncts to the cytological early diagnosis of ESCC
引文
Albertson DG,Collins C,McCormick F,Gray JW.(2003)Chromosome aberrations in solid rumors.Nat Genet.34:369-376.
    Andrejevic-Blant S,Osterheld MC,Caron L,Ballini JP,Monnier P.(2006)Deoxyribonucleic acid content as an indicator of progression of squamous cell carcinogenesis in the esophagus:comparative analysis on imprint-cytospin and tissue section preparation.Cancer Detect Prev.30:276-83
    Aujesky R,Hajduch M,Neoral C,Kr(?)l V,L'ubusska L,Bohanes T,Klein J,Vrba R,Drac P.(2005)p53--prognostic factor of malignant transformation of Barrett's esophagus.Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub.149:141-144
    Bergman J,Reznichek RC,Rajfer J.(2008)Surveillance of patients with bladder carcinoma using fluorescent in-situ hybridization on bladder washings.BJU Int.101:26-29
    Beuzen F,Dubois S,Flejou JF.(2000)Chromosomal numerical aberrations are frequent in oesophageal and gastric adenocarcinomas:a study using in-situ hybridizaiton.Histopathology.37:241-249
    Bitzer M,Stahl M,Arjumand J,Rees M,Klump B,Heep H,Gabbert HE,Sarbia M.(2003)C-myc gene amplification in different stages of oesophageal squamous cell carcinoma:prognostic value in relation to treatment modality.Anti-cancer Res.23:1489-1493
    Cai YC,So CK,Nie AY,Song Y,Yang GY,Wang LD,Zhao X,Kinzy TG,Yang CS.(2007)Characterization of genetic alteration patterns in human esophageal squamous cell carcinoma using selected microsatellite markers spanning multiple loci.Int J Oncol.30:1059-1067
    Cao W,Chen X,Dai H,Wang H,Shen B,Chu D,McAfee T,Zhang ZF.(2004)Mutational spectra of p53 in geographically localized esophageal squamous cell carcinoma groups in China.Cancer.101:834-844
    Chatrath P,Scott IS,Morris LS,Davies RJ,Bird K,Vowler SL,Coleman N.(2006)Immunohistochemical estimation of cell cycle phase in laryngeal neoplasia.Br J Cancer.95:314-21
    Daniely M,Rona R,Kaplan T,Olsfanger S,Elboim L,Freiberger A,Lew S,Leibovitch I.(2007)Combined morphologic and fluorescence in situ hybridization analysis of voided urine samples for the detection and follow-up of bladder cancer in patients with benign urine cytology.Cancer.111:517-524
    Dawsey SM,Fleischer DE,Wang GQ,Zhou B,Kidwell JA,Lu N,Lewin KJ,Roth MJ,Tio TL,Taylor PR.(1998)Mucosal iodine staining improves endoscopic visualization of squamous dysplasia and squamous cell carcinoma of the esophagus in Linxian,china.Cancer,83:220-231
    Dey,P.(2004)Aneuploidy and malignancy:an unsolved equation.J Clin Pathol.57:1245-9
    Doak SH,Jenkins GJ,Parry EM,D'Souza FR,Griffiths AP,Toffazal N,Shah V,Baxter JN,Parry JM.(2003)Chromosome 4 hyperploidy represents an early genetic aberration in premalignant Barrett's oesophagus.Gut.52:623-628
    Duesberg P,Li R,Rasnick D,Rausch C,Wilier A,Kraemer A,Yerganian G,Hehlmann R.(2000)Aneuploidy precedes and segregates with chemical carcinogenesis.Cancer Genet Cytogenet.119:83-93
    Freeman A,Morris LS,Mills AD,Stoeber K,Laskey RA,Williams GH,Coleman N.(1999)Minichromosome maintenance proteins as biological markers of dysplasia andmalignancy.Clin Cancer Res.5:2121-2132
    Falkenback D,Nilbert M,Oberg S,Johansson J.(2008)Prognostic value of cell adhesion in esophageal adenocarcinomas.Dis Esophagus.21:97-102
    Gerdes J,Lemke H,Baisch H,Wacker HH,Schwab U,Stein H.(1984)Cell cycle analysis of a cell proliferation-associated human nuclear antigen defined by the monoclonal antibody Ki67.J Immunol.133:1710-1715
    Gerdes J,Li L,Schlueter C,Duchrow M,Wohlenberg C,Gerlach C,Stahmer I,Kloth S,Brandt E,Flad HD.(1991)Immunobiochemical and molecular biologic characterization of the cell proliferation-associated nuclear antigen that is defined by monoclonal antibody Ki67.Am J Pathol.138:867-873
    Gerdes J,Schwab U,Lemke H,Stein H.(1983)Production of a mouse monoclonal antibody reactive with human nuclear antigen associated with cell proliferation.Int J Cancer.31:13-20
    Gibault L,Metges JP,Conan-Charlet V,Lozac'h P,Robaszkiewicz M,Bessaguet C,Lagarde N,Volant A.(2005)Diffuse EGFR staining is associated with reduced overall survival in locally advanced oesophageal squamous cell cancer.Br J Cancer 93:107-115
    Going JJ,Keith WN,Neilson L,Stoeber K,Stuart RC,Williams GH.(2002)Aberrant expression of minichromosome maintenance proteins 2 and 5,and Ki-67 in dysplastic squamous oesophageal epithelium and Barrett's mucosa.Gut.50:373-377.
    Gotoh M,Takiuchi H,Kawabe S,Ohta S,Kii T,Kuwakado S,Katsu K.(2007)Epidermal growth factor receptor is a possible predictor of sensitivity to chemoradiotherapy in the primary lesion of esophageal squamous cell carcinoma.Jpn J Clin Oncol.37:652-657
    Grothey A,Hashizume R,Sahin AA,McCrea PD.(2000)Fascin,an actinbundling protein associated with cell motility,is upregulated in hormone receptor negative breast cancer.Br J Cancer 83:870-873
    Heselmeyer-Haddad K,Sommerfeld K,White NM,Chaudhri N,Morrison LE,Palanisamy N,Wang ZY,Auer G,Steinberg W,Ried T.(2005)Genomic amplification of the human telomerase gene(TERC)in pap smears predicts the development of cervical cancer.Am J Pathol.166:1229-1238
    Huang JX,Yan W,Song ZX,Qian RY,Chen P,Salminen E,Toppari J.(2005)Relationship between proliferative activity of cancer cells and clinicopathological factors in patients with esophageal squamous cell carcinoma.World J Gastroenterol.11:2956-9
    Itami A,Shimada Y,Watanabe G,Imamura M.(1999)Prognostic value of p27(Kipl)and CyclinDl expression in esophageal cancer.Oncology 57:311-317
    Jin Y,Jin C,Law S,Chu KM,Zhang H,Strombeck B,Yuen AP,Kwong YL.(2004)Cytogenetic and fluorescence in situ hybridization characterization of clonal chromosomal aberrations and CCND1 amplification in esophageal carcinomas.Cancer Genet Cytogenet.148:21-28
    Jung SY,Han W,Lee JW,Ko E,Kim E,Yu JH,Moon HG,Park IA,Oh DY,Im SA,Kim TY,Hwang KT,Kim SW,Noh DY.(2009)Ki-67 expression gives additional prognostic information on St.Gallen 2007 and Adjuvant! Online risk categories in early breast cancer.Ann Surg Oncol.2009 May;16(5):1112-21
    Kaifi JT,Yekebas EF,Schurr P,Obonyo D,Wachowiak R,Busch P,Heinecke A,Pantel K,Izbicki JR.(2005)Tumor-cell homing to lymph nodes and bone marrow and CXCR4 expression in esophageal cancer.J Natl Cancer Inst.97:1840-1847
    Kato H,Miyazaki T,Fukai Y,Nakajima M,Sohda M,Takita J,Masuda N,Fukuchi M,Manda R,Ojima H,Tsukada K,Asao T,Kuwano H.(2003)A new proliferation marker,minichromosome maintenance protein 2,is associated with tumor aggressiveness in esophageal squamous cell carcinoma.J Surg Oncol.84:24-30
    Kawakubo H,Ozawa S,Ando N,Kitagawa Y,Mukai M,Ueda M,Kitajima M.(2005)Iterations of p53,cyclin Dl and pRB expression in the carcinogenesis of esophageal squamous cell carcinoma.Oncol Rep.14:1453-1459
    Koishi K,Yoshikawa R,Tsujimura T,Hashimoto-Tamaoki T,Kojima S,Yanagi H,Yamamura T,Fujiwara Y.(2006)Persistent CXCR4 expression after preoperative chemoradiotherapy predicts early recurrence and poor prognosis in esophageal cancer.World J Gastroenterol.12:7585-7590
    Kops GJ,Weaver BA,Cleveland DW.(2005)On the road to cancer:aneuploidy and the mitotic checkpoint.Nat Rev Cancer..5:773-85
    Kou YL,Zhang JR,Zhou Q,Zhao YF,Chen GD.(2004)Analysis of p53 gene mutation in esophageal exfoliative cells Zhonghua Zhong Liu Za Zhi.26:349-352
    Levy MJ,Clain JE,Clayton A,Hailing KC,Kipp BR,Rajan E,Roberts LR,Root RM,Sebo TJ,Topazian MD,Wang KK,Wiersema MJ,Gores GJ.(2007)Preliminary experience comparing routine cytology results with the composite results of digital image analysis and fluorescence in situ hybridization in patients undergoing EUS-guided FNA.Gastrointest Endosc.66:483-90
    Lu N,Hu N,Li WJ,Roth MJ,Wang C,Su H,Wang QH,Taylor PR,Dawsey SM.(2003)Microsatellite alterations in esophageal dysplasia and squamous cell carcinoma from laser capture microdissected endoscopic biopsies.Cancer Lett.189:137-145
    Lung JC,Chu JS,Yu JC,Yue CT,Lo YL,Shen CY,Wu CW.(2002)Aberrant expression of cell-cycle regulator cyclin Dl in breast cancer is related to chromosomal genomic instability.Genes Chromosomes Cancer.34:276-284
    Manoel-Caetano Fda S,Borim AA,Caetano A,Cury PM,Silva AE.(2004)Cytogenetic alterations in chagasic achalasia compared to esophageal carcinoma.Cancer Genet Cytogenet.149:17-22
    Mohan V,Ponnala S,Reddy HM,Sistla R,Jesudasan RA,Ahuja YR,Hasan Q.(2007)Chromosome 11 aneusomy in esophageal cancers and precancerous lesions--an early event in neoplastic transformation:an interphase fluorescence in situ hybridization study from south India.World J Gastroenterol.13:503-508
    Moskaluk CA,Hu J,Perlman EJ.(1998)Comparative genomic hybridisation of esophageal and gastroesophageal adenocarcinomas shows consensus areas of DNA gain and loss.Genes Chromosomes Cancer.22:305-11
    Nakamura T,Hayashi K,Ota M,Ide H,Takasaki K,Mitsuhashi M.(2004)Expression of p21(Waf1/Cip1)predicts response and survival of esophageal cancer patients treated by chemoradiotherapy.Dis Esophagus.2004 17:315-321
    Nita ME,Nagawa H,Tominaga O,Tsuno N,Hatano K,Kitayama J,Tsuruo T,Domene CE,Muto T.(1999)p21Wafl/Cipl expression is a prognostic marker in curatively resected esophageal squamous cell carcinoma,but not p27Kipl,p53,or Rb.Ann Surg Oncol.6:481-488
    Okumura H,Natsugoe S,Matsumoto M,Mataki Y,Takatori H,Ishigami S,Takao S, Aikou T.(2005)The predictive value of p53,p53R2,and p21 for the effect of chemoradiation therapy on oesophageal squamous cell carcinoma.Br J Cancer.92:284-289
    Pan QJ,Roth MJ,Guo HQ,Kochman ML,Wang GQ,Henry M,Wei WQ,Giffen CA,Lu N,Abnet CC,Hao CQ,Taylor PR,Qiao YL,Dawsey SM.(2008)Cytologic detection of esophageal squamous cell carcinoma and its precursor lesions using balloon samplers and liquid-based cytology in asymptomatic adults in Llinxian,China.Acta Cytol.52:14-23
    Parkin DM,Bray FI,Devesa SS.(2001)Cancer burden in the year 2000.The global picture.Eur J Cancer.37:S4-S66
    Peters FP,Krishnadath KK,Rygiel AM,Curvers WL,Rosmolen WD,Fockens P,Ten Kate FJ,van Baal JW,Bergman JJ.(2007)Stepwise radical endoscopic resection of the complete esophagus with early neoplasia successfully eradicates pre-existing genetic abnormalities.Am J Gastroenterol.102:1853-1861
    Reid BJ,Prevo LJ,Galipeau PC,Sanchez CA,Longton G,Levine DS,Blount PL,Rabinovitch PS.(2001)Predictors of progression in Barrett's esophagus II:baseline 17p(p53)loss of heterozygosity identifies a patient subset at increased risk for neoplastic progression.Am J Gastroenterol.96:2839-2848
    Riegman PH,Vissers KJ,Alers JC,Geelen E,Hop WC,Tilanus HW,van Dekken H.(2001)Genomic alterations in malignant transformation of Barrett's esophagus.Cancer Res.61:3164-3170
    Roth MJ,Liu SF,Dawsey SM,Zhou B,Copeland C,Wang GQ,Solomon D,Baker SG,Giffen CA,Taylor PR.(1997)Cytologic detection of esophageal squamous cell carcinoma and precursor lesions using balloon and sponge samplers in asymptomatic adults in Linxian,China.Cancer.80:2047-2059
    Sarbia M,Ott N,Piihringer-Oppermann F,Briicher BL.(2007)The predictive value of molecular markers(p53,EGFR,ATM,CHK2)in multimodally treated squamous cell carcinoma of the oesophagus.Br J Cancer.97:1404-1408
    Savic S,Glatz K,Schoenegg R,Spieler P,Feichter G,Tamm M,Bubendorf L.(2006)Multitarget fluorescence in situ hybridization elucidates equivocal lung cytology.Chest.129:1629-1635
    Shibata H,Matsubara O,Wakiyama H,Tanaka S.(2001)The role of cyclin-dependent kinase inhibitor p27 in squamous cell carcinoma of the esophagus.Pathol Res Pract.197:157-164
    Shi ST,Yang GY,Wang LD,Xue Z,Feng B,Ding W,Xing EP,Yang CS.(1999)Role of p53 gene mutations in human esophageal carcinogenesis:results from immunohistochemical and mutation analyses of carcinomas and nearby non-cancerous lesions.Carcinogenesis 20:591-597
    Simada Y,Imamura M,Watanabe G,et al.Prognostic factors of oe2 sophageal cell carcinoma from the perspective of molecular biology.Br J Cancer,1999 80(8):1281
    Sirieix PS,O'Donovan M,Brown J,Save V,Coleman N,Fitzgerald RC.(2003)Surface Expression of Minichromosome Maintenance Proteins Provides a Novel Method for Detecting Patients at Risk for Developing Adenocarcinoma in Barrett's Esophagus Clin Cancer Res.9:2560-2566
    Sokolova IA,Hailing KC,Jenkins RB,Burkhardt HM,Meyer RG,Seelig SA,King W.(2000)The Development of a Multitarget,Multicolor Fluorescence in Situ Hybridization Assay for the Detection of Urothelial Carcinoma in Urine.J Mol Diagn.2:116-23
    Solomon D,Davey D,Kurman R,Moriarty A,O'Connor D,Prey M,Raab S,Sherman M,Wilbur D,Wright T Jr,Young N;Forum Group Members;Bethesda 2001 Workshop.(2002)The 2001 Bethesda system:terminology for reporting results of cervical cytology.JAMA,287:2114-2119
    Sunada F,Itabashi M,Ohkura H,Okumura T.(2005)p53 negativity,CDC25B positivity,and metallothionein negativity are predictors of a response of esophageal squamous cell carcinoma to chemoradiotherapy.World J Gastroenterol.11:5696-5700
    Swierczynski SL,Maitra A,Abraham SC,Iacobuzio-Donahue CA,Ashfaq R,Cameron JL,Schulick RD,Yeo CJ,Rahman A,Hinkle DA,Hruban RH,Argani P.(2004)Analysis of novel tumor markers in pancreatic and biliary carcinomas using tissue microarrays.Hum Pathol.35:357-366
    Tada K,Oka M,Hayashi H,Tangoku A,Oga A,Sasaki K.(2000)Gains of 8q23-qter and 20q and loss of 11q22-qter in esophageal squamous cell carcinoma associated with lymph node metastasis.Cancer.88:268-273
    Todorov IT,Werness BA,Wang HQ,Buddharaju LN,Todorova PD,Slocum HK,Brooks JS,Huberman JA.(1998)HsMCM2/BM28:a novel proliferation marker for human tumors and normal tissues.Lab Invest.78:73-78
    Tokugawa T,Sugihara H,Tani T,Hattori T.(2002)Modes of silencing of p16 in development of esophageal squamous cell carcinoma.Cancer Res.62:4938-4944
    Tselepis C,Morris CD,Wakelin D,Hardy R,Perry I,Luong QT,Harper E,Harrison R,Attwood SE,Jankowski JA.(2003)Upregulation of the oncogene c-myc in Barrett's adenocarcinoma:induction of c-myc by acidified bile acid in vitro.Gut.52:174-80
    Tye,BK.MCM proteins in DNA replication.(1999)Annu.Rev.Biochem.68:649-686
    van Dekken H,Vissers CJ,Tilanus HW,Tanke HJ,Rosenberg C.(1999)Clonal analysis of a case of multifocal oesophageal(Barrett's)adenocarcinoma by comparative genome hybridisation.J Pathol.188:263-266
    Walch AK,Zitzelsberger HF,Bruch J,Keller G,Angermeier D,Aubele MM,Mueller J,Stein H,Braselmann H,Siewert JR,Hofler H,Werner M.(2000)Chromosomal imbalances in Barrett's adenocarcinoma and the metaplasia-dysplasia-carcinoma sequence.Am J Pathol.156:555-566
    Wang GQ,Jiao GG,Chang FB,Fang WH,Song JX,Lu N,Lin DM,Xie YQ,Yang L.(2004)Long term results of operation for 420patients with early squamous cell esophageal carcinoma discovered by screening.Annals of Thoracic Surgey.77:1740-1744
    Wang LD,Qin YR,Fan ZM,Kwong D,Guan XY,Tsao GS,Sham J,Li JL,Feng XS.(2006)Comparative genomic hybridization:comparison between esophageal squamous cell carcinoma and gastric cardia adenocarcinoma from a high-incidence area for both cancers in Henan,northern China.Dis Esophagus.19:459-67
    Wang XL,Zhang CM,Shi LY,Yu HP,Xu SQ.(2004)Significance of p53 gene mutation and P53 protein expression abnormality on the prognosis of esophageal cancer:a meta-analysis study.Zhonghua Liu Xing Bing Xue Za Zhi.25:769-774
    Wongsurawat VJ,Finley JC,Galipeau PC,Sanchez CA,Maley CC,Li X,Blount PL,Odze RD,Rabinovitch PS,Reid BJ.(2006)Genetic mechanisms of TP53 loss of heterozygosity in Barrett's esophagus:implications for biomarker validation.Cancer Epidemiol Biomarkers Prev.15:509-516
    Xue LY,Hu N,Song YM,Zou SM,Shou JZ,Qian LX,Ren LQ,Lin DM,Tong T,He ZG,Zhan QM,Taylor PR,Lu N.(2006)Tissue microarray analysis reveals a tight correlation between protein expression pattern and progression of esophageal squamous cell carcinoma.BMC Cancer.22:296-301
    Yamaki H,Sasano H,Ohashi Y,Shizawa S,Shineha R,Satomi S,Nagura H.(2001)Alteration of X and Y chromosomes in human esophageal squamous cell carcinoma.Anticancer Res.21:985-990
    Yang Y,Fruehauf J,Xiang S,Li CJ.(2006) Genomic Instability in Precancerous Lesions before Inactivation of Tumor Suppressors p53 and APC in Patients.Cell Cycle.5:1443-1447
    Yen CC,Chen YJ,Lu KH,Hsia JY,Chen JT,Hu CP,Chen PM,Liu JH,Chiou TJ,Wang WS,Yang MH,Chao TC,Lin CH.(2003) Genotypic analysis of esophageal squamous cell carcinoma by molecular cytogenetics and real-time quantitative polymerase chain reaction.Int J Oncol.23:871-881
    Yoder BJ,Skacel M,Hedgepeth R,Babineau D,Ulchaker JC,Liou LS,Brainard JA,Biscotti CV,Jones JS,Tubbs RR.(2007) Reflex UroVysion testing of bladder cancer surveillance patients with equivocal or negative urine cytology:a prospective study with focus on the natural history of anticipatory positive findings.Am J Clin Pathol.127:295-301
    Zhang H,Xu L,Xiao D,Xie J,Zeng H,Cai W,Niu Y,Yang Z,Shen Z,Li E.(2006)Fascin is a potential biomarker for early-stage oesophageal squamous cell carcinoma.J Clin Pathol.59:958-964
    Zuo L,Ling P,Qi F,Wang J,Ding Z,Guo J,Liu J.(2000)Flow cytometric DNA analyses of epithelial dysplasia of the esophagus.Anal Quant Cytol Histol.22:175-177
    黄俊星,宋振湘,钱荣誉,徐光炜.(2003)细胞周期调节蛋白在食管鳞癌组织中的表达.癌症.22:277-278
    靳玉兰,张伟,刘伯齐,王洪平,韩志楷,韩双廷,曲平,李茉,丁镇伟,林培中.(2001)食管原位癌及癌前病变组织中Ki67、p53、iNOS的异常表达.中华肿瘤杂志,23:129-131
    康维,姚汉清,房丽丽,蔡岩,韩亚铃,徐听,张钰,贾雪梅,王明荣.(2009)食管鳞癌3、8、10、20和Y染色体的非整倍性分析.遗传.31:255-260
    刘卫梅.(2007)p21在食管癌组织中的表达及临床意义.肿瘤基础与临床.20:173-174
    马正中,阚秀,刘树范.(2000)河南:河南科学技术出版社.诊断细胞病理学.177-180
    秦艳茹,王立东,邝丽芸,高珊珊,关新元,庄则豪,范宗民,邓文,胡亮.(2005)利用比较基因组杂交技术分析食管癌组织中染色体基因组的变化特征.中华病理杂志.34:80-83
    全国肿瘤防治研究办公室,全国肿瘤登记中心,卫生部疾病预防控制局编.(2008)北京:中国协和医科大学出版社中国肿瘤登记年报 2004.41-85
    滕梁红,吕宁,赵晓航,薛丽燕,刘芳,林冬梅,王卉心,何祖根,谢永强,刘秀云.(2005)组织芯片技术检测食管癌前病变相关预警标志物.世界华人消化杂志.13:6-9
    王国清,郝常青,赖少清.(2002)贲门癌高发位点的内镜研究.中华肿瘤杂志.24:381-383
    吴旻.肿瘤遗传学.北京:科学出版社.2004
    姚汉清,贺舜,吴玉鹏,王小春,韩亚玲,徐昕,蔡岩,王贵齐,王明荣.(2008)多色荧光原位杂交应用于食管鳞状细胞癌早期诊断的探讨.癌症,27:1137-1143
    张天宝,张钰,蔡岩,韩亚铃,徐昕,王明荣,贾雪梅.(2007)食管癌染色体畸变及其与临床病理参数相关性研究.安徽医科大学学报.42:603-608
    周彬,丁镇伟,郭黎平,潘秦镜,高峰,王继信,林培中.(1999)细胞图像分析预测食管上皮增生转归的研究.中华肿瘤杂志.21:439-443
    左连富,林培中,齐凤蔓,张林西,郭建文,刘江惠.(2002)食管癌前细胞DNA含量及多基因表达的定量检测.中华肿瘤杂志.24:30-33