细胞周期依赖性蛋白激酶1（Cdk1/p34 cdc2）对喉癌术后局部复发的预测价值

目的：检测喉鳞状细胞癌及癌旁切缘组织中p53、p21和Cdk1/p34cdc2蛋白表达及其与患者预后的关系。方法应用免疫组化SP法检测85例喉鳞状细胞癌及癌旁切缘组织中p53、p21和Cdk1/p34cdc2蛋白的表达,并收集临床病理资料进行随访。结果在随访2年以上的喉鳞状细胞癌中14例出现局部复发(复发组),71例未复发(未复发组)。在喉鳞状细胞癌和癌切缘组织中p53蛋白阳性率分别为60．0%和36．5%;p21蛋白阳性率分别为38．8%和21．2%;Cdk1/p34cdc2蛋白阳性率分别为70．6%和29．4%。喉鳞状细胞癌复发组和未复发组的切缘组织中p53蛋白阳性率分别为71．4%和29．6%( P=0．003),切缘组织中p21蛋白阳性率分别为50．0%和15．5%(P=0．004),切缘组织中 Cdk1/p34cdc2蛋白阳性率分别为57．1%和23．9%(P=0．013)。癌旁切缘组织中p53与p21及Cdk1/p34cdc2蛋白表达之间无相关性(P均>0．05)。结论 p53、p21和Cdk1/p34cdc2蛋白在喉鳞状细胞癌的发生、发展中可能起重要作用;癌旁切缘组织中p53、p21和Cdk1/p34cdc2过表达与喉鳞状细胞癌局部复发密切相关。     

p21 Ras和表皮生长因子受体蛋白在喉鳞状细胞癌组织中的表达

目的探讨p21 Ras及表皮生长因子受体在喉鳞状细胞癌发病机制中的表达及相关性分析。方法选取50例喉鳞状细胞癌组织(实验组)及10例正常喉黏膜组织(对照组),应用免疫组织化学方法检测两组组织中p21 Ras和表皮生长因子受体的蛋白表达,并进行统计学分析。结果与对照组相比,p21 Ras蛋白在喉鳞状细胞癌组织中的阳性表达率显著升高(分别为40.0%和72.0%,P0.05),且与喉鳞状细胞癌临床分期、分化程度及淋巴结转移密切相关(P0.05);与对照组相比,EGFR在喉鳞状细胞癌组织中的表达显著升高(分别为20.0%和76.0%,P0.05),且与喉鳞状细胞癌淋巴结转移密切相关(P0.05),而与性别、肿瘤发生部位、分化程度、临床分期无关(P0.05)。相关性分析结果显示,EGFR与p21 Ras表达呈正相关(r=0.81,P0.05)。结论 p21 Ras和EGFR在喉鳞状细胞癌组织中的高阳性率可作为喉鳞状细胞癌发生的预测指标,在伴有淋巴结转移组的强阳性表达提示二者在喉鳞状细胞癌发展过程中起重要作用。     

bcl-2、ki-67和p53在喉鳞状细胞癌中的表达及临床意义

目的:探讨bcl-2、ki-67和p53在喉鳞状细胞癌中的表达及临床意义。方法:采用SP免疫组织化法检测60例喉鳞状细胞癌患者bcl-2、ki-67和p53的表达,并结合临床病理因素进行分析。结果:喉鳞状细胞癌中bcl-2、P53、Ki-67的阳性表达率分别为60.0%(36/60)、53.3%(32/60)、30.0%(18/60)。喉鳞状细胞癌患者的bcl-2、P53和Ki-67阳性表达与组织学分级及淋巴结转移密切相关,组织学分级越低,伴有淋巴结转移的喉鳞状细胞癌患者的bcl-2、P53和Ki-67阳性表达率明显高于其他组(P0.05),但与年龄、肿瘤大小并无显著相关性(P0.05)。结论:bcl-2、ki-67和p53是评价喉鳞状细胞癌预后的较好指标,三者联合检测有助于判断喉鳞状细胞癌的病理临床特征。     

食管鳞状细胞癌组织中核干细胞因子、表皮生长因子和表皮生长因子受体mRNA的表达

目的 探讨食管鳞状细胞癌组织中核干细胞因子(NS),表皮生长因子(EGF)和表皮生长因子受体(EGFR)mRNA的表达及其相互关系. 方法 采用原位杂交技术检测62例食管鳞状细胞癌组织,31例癌旁不典型增生组织及62例正常食管黏膜组织中NS、EGF和EGFR mRNA的表达阳性率,并分析食管鳞状细胞癌患者不同临床病理参数间NS、EGF和EGFR mRNA的表达阳性率及三者之间的相关性. 结果 正常食管黏膜组织、癌旁不典型增生组织及食管鳞状细胞癌组织中NS mRNA的阳性表达率分别为21.0%(13/62)、25.8%(8/31)和69.4%(43/62);EGF mRNA阳性表达率分别是40.3%(25/62)、48.4%(15/31)和77.4%(48/62);EGFR mRNA阳性表达率分别是30.6%(19/62),45.2%(14/31)和75.8%(47/62).食管鳞状细胞癌组织中NS、EGF和EGFR mRNA的阳性表达率与肿瘤的组织学分级,浸润深度及淋巴结转移有关(均P<0.05).食管鳞状细胞癌组织中NS mRNA表达与EGFmRNA和EGFR mRNA的表达呈正相关(分别为r=0.394,r=0.604,P<0.05). 结论 食管鳞状细胞癌组织中NS mRNA与EGF mRNA和EGFR mRNA的表达呈正相关,NS mRNA、EGF mRNA和EGFR mRNA在食管癌的发生、发展过程中可能起重要作用.     

临床喉鳞癌组织标本中G3BP和CD44v6的检测及意义北大核心CSCD

目的研究RNA结合的Ras-GAP酶激活蛋白SH3结合蛋白(G3BP)、肿瘤干细胞标志物CD44v6蛋白与喉鳞状细胞癌(喉鳞癌)发生发展的关系,探讨G3BP、CD44v6蛋白与血管新生的相关性。方法收集56例喉鳞癌患者癌组织及对应癌旁组织,采用Western blot法检测喉鳞癌患者癌组织及对应癌旁组织中G3BP、CD44v6蛋白水平,使用免疫组织化学染色法分别检测G3BP、CD44v6、血管内皮生长因子A(VEGF-A)的表达,并分析G3BP、CD44v6表达与喉鳞癌患者临床病理特征的关系及与VEGF-A表达的相关性;采用Ⅷ因子相关抗原(FⅧAg)免疫组织化学染色确定微血管密度(MVD),观察G3BP、CD44v6表达与MVD之间的关系。结果喉鳞癌患者癌组织中G3BP蛋白、CD44v6蛋白水平均明显高于对应癌旁的正常组织。喉鳞癌患者癌组织中的G3BP、CD44v6、VEGF-A表达阳性率分别为58.9%、53.6%、46.4%,均明显高于对应的癌旁正常组织。喉鳞癌患者癌组织中G3BP、CD44v6蛋白表达阳性率与临床分期、复发或转移及淋巴结转移相关,但与年龄和肿瘤大小无关。经Pearson相关性分析,喉鳞癌患者癌组织中G3BP、CD44v6蛋白的表达与VEGF-A表达呈正相关(r=0.741,r=0.756);喉鳞癌患者癌组织中G3BP与CD44v6蛋白阳性表达病例的MVD值均明显高于癌旁正常组织中MVD值;癌组织中G3BP与CD44v6蛋白阴性表达病例MVD值与癌旁正常组织中MVD值比较无明显差异。结论喉鳞癌组织G3BP、CD44v6蛋白高表达,可能通过促进VEGF-A的表达来诱发肿瘤微血管的新生,促进喉鳞癌侵袭、转移。     

细胞周期素E和p53在乳腺癌中表达及意义

目的 探讨细胞周期素E(cyclin E)和p53在乳腺癌中表达的意义和相瓦关系.方法 采用免疫组化方法 (SP法)观察82例乳腺癌、15例乳腺良性肿瘤与15例正常乳腺组织的eyclin E和p53的表达.结果 正常乳腺组织和良性肿瘤中cyclin E和p53无表达,乳腺癌组织中cyclin E和p53的阳性表达率分别为56.10%和60.98%(P<0.05),cyclin E和p53的阳性表达与乳腺癌的临床分期密切相关(P<0.05);cyclin E和p53在乳腺癌的阳性表达具有相关(P<0.05),共同阳性表达的患者5年生存率明显低于阴性者(P<0.05).结论 cyclin E在乳腺癌中的表达与p53的表达呈明显的一致性,并在肿瘤的进展中起一定的作用,cyclin E和p53在乳腺癌中的共同阳性表达可作为乳腺癌预后的有效指标.     

胃肠道间质瘤中p53和bcl-2表达与预后的关系

目的探讨细胞凋亡相关基因p53和bc l-2在胃肠道间质瘤(gastrointestinal strom al tumors,G ISTs)中的表达与预后关系。方法对194例G ISTs构建组织微阵列(TMA),采用免疫组化EnV ision法检测G ISTs组织中p53和bc l-2基因蛋白的表达。结果在p53 TMA中,184例可评估(94.8%),在bc l-2 TMA中181例可评估(93.3%)。p53和bc l-2基因蛋白阳性率分别为34.8%和59.1%。p53蛋白阳性表达率与肿瘤大小、NIH分级、肿瘤部位、坏死、细胞密集程度、核分裂象和转移复发有关。bc l-2阳性表达率与肿瘤大小、NIH分级、肿瘤部位、坏死和黏膜受累及有关。p53蛋白阳性和阴性表达者的5年生存率分别为40.1%和76.5%,两者比较差异有显著性(P<0.01)。bc l-2阳性和阴性表达者的5年生存率分别为55.1%和76.2%,两者比较差异有显著性(P<0.05)。p53蛋白阳性组和阴性组与bc l-2蛋白的阳性表达差异有显著性(P<0.01)。结论p53、bc l-2表达与G ISTs预后有关,p53、bc l-2可作为判断G ISTs预后的标志物。     

肺小细胞癌p53基因分析与mdm-2基因蛋白表达的关系

目的:探讨肺小细胞癌(SCLC)的p53基因改变与mdm-2基因蛋白表达的关系.方法:应用免疫组化LSAB法和聚合酶链反应-单链构象多态性(PCR-SSCP)分析的方法,对14例SCLC的石蜡切片组织进行研究.结果:免疫组化染色p53蛋白阳性9例,阳性率为64.3%(9/14);mdm-2蛋白阳性4例,阳性率为28.6%(4/14).p53基因第5、6、7、8外显子,突变率分别为21.4%(3/14);14.3(2/14);14.3%(2/14);7.1%(1/14),总突变率为57.1%(8/14).结论:SCLC中存在较高的p53基因突变率和mdm-2基因蛋白的明显表达.     

p57kip2、cyclin D1在食管癌中的表达及意义

目的研究p57kip2在食管癌中的表达及意义,探讨其与cyclin D1在食管癌中表达的相关性。方法采用免疫组织化学SP法检测50例食管癌组织及15例正常食管上皮组织中p57kip2、cyclin D1的表达情况;应用流式细胞术对这些组织的DNA含量和细胞周期分布进行分析。结果免疫组化:p57kip2蛋白在食管癌组织中阳性表达率(34.0%)低于正常组织(80%)(P<0.01),与食管癌组织分化程度、浸润深度和淋巴结转移情况有关(P<0.05);cyclin D1蛋白在食管癌组织中的阳性表达率(64.0%)高于正常组织(13.33%)(P<0.01),与浸润深度和淋巴结转移情况有关(P<0.05),与食管癌组织分化程度无关(P>0.05)。p57kip2和cyclin D1之间表达负相关(r=-0.429,P<0.01)。流式细胞术:与正常组织相比,癌组织中DNA含量增高,异倍体细胞增加;G0/G1期细胞减少,而S期和G2/M期细胞增多,增殖指数(PI)高于正常组织。结论p57kip2蛋白低表达可能与食管癌的发生发展有关;p57kip2与cyclin D1联合检测可作为评估食管癌恶性程度指标。     

益气解毒方对TgN(p53mt-LMP1)/HT小鼠鼻咽黏膜上皮细胞周期相关蛋白的影响

目的 研究益气解毒方对TgN (p53mt-LMP l)/HT转基因小鼠鼻咽黏膜上皮细胞周期相关蛋白p53、p21、CDK2和cyclinD1的影响.方法 TgN (p53mt-LMP1)/HT转基因小鼠随机分为3组,分别为模型组(MG)、益气解毒方干预组(YG)、维甲酸阳性对照组(TG),每组24只;同品系C57BL/6J鼠24只作对照组(CG).各组均用诱癌剂二亚硝基哌嗪进行处理(每次剂量73 mg/kg,每周2次,共28次).采用免疫组织化学染色法检测小鼠鼻咽黏膜上皮细胞周期相关蛋白p53、p21、CDK2和cyclinD1的表达水平,并采用Western blot法进行验证.结果 YG组、TG组及CG组p53蛋白和cyclinD1蛋白表达均显著低于MG组(P＜0.05);YG、TG及CG组p21蛋白表达量高于MG组,差异有统计学意义(P＜0.05);各组CDK2蛋白表达量差异无统计学意义.结论 益气解毒方逆转鼻咽黏膜癌前病变的发展可能与下调cyclin D1蛋白和上调p21蛋白表达有关.     

The expression of ribonuclear protein IMP3 in laryngeal carcinogenesis

ObjectiveThe aim of the study was to investigate the expression of ribonuclear protein IMP3 in laryngeal carcinogenesis, together with other biomarkers of carcinogenesis (Ki-67, p53 and cyclin D1), and to evaluate their predictive values.MethodsThe study included 153 patients divided into three groups: 68 operated for primary invasive laryngeal squamous cell carcinoma (LSCC); 41 with precancerous lesions of atypical and abnormal hyperplasia; 44 with hyperplastic laryngeal nodule without atypia. Tissue microarray technique was used for immunohistochemical analysis.ResultsAll markers showed statistically significant differences between the three groups. The percentage of IMP3 positive cells is statistically significantly higher in LSCC group in comparison to precancerosis and control group. The percentage of Ki-67 positive cells is statistically significantly higher in LSCC group in comparison to precancerosis and control group. The percentage of p53 positive cells in LSCC group is statistically significantly higher than the control group and higher, but not statistically significant, than the precancerosis group. The percentage of cyclin D1 positive cells is statistically significantly higher in LSCC group than in precancerosis group and higher, but not statistically significant, than in the control group. All analyzed markers have good predictive values (AUC > 0.6), but the percentage of IMP3 positive cells is the only statistically significant marker in predicting whether the patient has LSCC or not.ConclusionExpression of Ki-67 and pronouncedly IMP3 generally follow the same pattern where control and precancerosis are similar and LSCC significantly differs, as opposed to p53 and cyclin D1. IMP3 expression increase possibly has an important diagnostic, therapeutic (in terms of the need for additional therapy after surgery) and prognostic value. Further studies on the exact molecular mechanisms behind it are, of course, needed.     

p53 Protein expression in laryngeal squamous cell carcinomas bearing wild-type and mutated p53 gene

We performed an immunohistochemical analysis to investigate the expression of p53 protein in a panel of 18 laryngeal squamous cell carcinomas, 15 primary tumours and three in relapse, previously analysed by us for the presence of p53 gene mutations. Dysplastic and/or normal surrounding mucosa was evaluated in 15 different tumours. The results of our study are the following: (1) expression of p53 protein was observed in one out of five tumours positive for p53 gene mutations (20%) and in 10 out of 13 (80%) negative cases; (2), p53 protein over-expression was frequently observed in normal and/or dysplastic mucosa surrounding either wild-type (7/11) or mutated p53 tumours (2/4); (3), p53 immunoreactive cells showed a pattern of distribution in normal and mildly/moderately dysplastic mucosa (basal layers), different from that in severely dysplastic mucosa (whole thickness). These data further support the hypothesis that p53 protein over-expression may be a marker of the earliest phases of multistep tumorigenesis in laryngeal squamous cell carcinoma.     

Survivin expression is a negative prognostic marker in laryngeal squamous cell carcinoma and is associated with p53 accumulation

AIMS: To investigate survivin expression in laryngeal squamous cell carcinoma (LSCC), its prognostic significance and relation to p53 status. Survivin is an inhibitor of apoptosis protein that is overexpressed in cancer. It has been implicated in both prevention of apoptosis and cell cycle regulation. It has been suggested that wild-type p53 represses survivin expression. METHODS AND RESULTS: Expression of survivin and p53 was analysed in 68 archival biopsy specimens of LSCC by immunohistochemistry. Survivin was detected in 67 of 68 LSCC cases; the proportion of survivin-positive cells varied from 8.2% to 100%. It was localized in the nucleus and/or cytoplasm of tumour cells. Of LSCC cases, 31.8% were p53+. The number of survivin-positive cells was significantly higher in the p53+ group. A high level of survivin expression and a supraglottic site of the tumour were two independent adverse prognostic factors in LSCC. CONCLUSIONS: Survivin is expressed in a varying proportion of cells in virtually all cases of LSCC. A high level of its expression predicts poor survival. Loss of wild-type p53 is a possible mechanism of survivin up-regulation in LSCC.     

MUS81基因在喉癌中的突变和表达

目的 探讨MUS81基因突变和表达与喉癌发生发展的相关性.方法 应用聚合酶链反应-单链构象多态性分析技术结合DNA测序检测分析了42例喉癌患者MUS81基因第9、10外显子的突变;应用半定量逆转录-PCR和Western印迹方法分析MUS81基因在喉癌组织中的表达情况.结果 42例喉癌、癌旁组织标本中,癌旁正常组织均无突变,喉癌组织标本中19例发生突变,占45.2%(19/42),11例喉癌组织第9外显子发现有突变,占26.2%(11/42),8 例喉癌组织第10外显子有突变,占19%(8/42),分析表明具有统计学意义(P<0.01).逆转录-PCR结果表明,42例喉癌中有17例MUS81基因mRNA低表达,占40.48%(17/42).Western印迹方法分析结果表明,42例喉癌中有17例MUS81蛋白质低表达,占40.48%(17/42),经统计学分析肿瘤组与对照组差异有统计学意义(P<0.01).分析表明MUS81基因突变与mRNA和蛋白质低表达有显著相关性(P<0.01).统计结果显示喉癌MUS81基因突变与TNM分期、年龄和淋巴结转移无相关性(P>0.05).MUS81基因低表达与TNM分期、年龄和淋巴结转移无相关性(P>0.05).结论 发现MUS81基因在喉癌组织中有突变发生及表达异常,提示MU81基因突变和表达异常可能是喉癌发生及发展的重要因素之一.     

p53基因突变与喉癌生物学行为的关系

目的　探讨 p5 3基因突变与喉癌生物学行为的关系。方法　应用聚合酶链反应 -单链构象多态性 (PCR- single strand conformation polymorphism,PCR- SSCP)银染技术结合 DNA直接测序 ,检测喉鳞癌新鲜或冰冻组织中 p5 3外显子 5～ 8的基因突变。结果　 - 、 - 喉鳞癌 p5 3基因突变率分别为6 9. 2 % (18/2 6 )和 85 .3% (2 9/34 ) ,P>0 .0 5 ;高、中、低分化程度的突变率依次为 5 2 .9% (9/17)、83.3%(2 0 /2 4)和 94.7% (18/19) ,P<0 .0 5 ;有、无颈淋巴结转移者突变率分别为 96 .4% (2 7/2 8)和 6 2 .5 % (2 0 /32 ) ,P<0 .0 1。从 47例 SSCP阳性标本中随机抽取 2 0例进行测序 ,19例测序为阳性 ,二者符合率为 95 %。结论　p5 3基因突变与病理分化程度及颈淋巴结转移有明显相关性。     

P53和Ki-67在喉鳞状细胞癌中的表达及与短期预后的关系

目的:探讨P53和Ki-67在喉鳞状细胞癌(laryngeal squamous cell carcinoma,LSCC)中的表达情况及临床病理参数和短期预后相关分析.方法:选择手术切除LSCC标本35例,肿瘤分期均为T3~4N0M0.采用SP法染色进行P53和Ki-67免疫组织化学标记,结合临床病理参数和2年随访结果及无病生存情况进行统计学分析.结果:LSCC标本P53阳性率为60.00%;2年复发率P53阳性患者76.19%高于P53阴性28.59%,差异具有统计学意义(P=0.013).年龄大小、是否抽烟、增殖指数高低及肿瘤不同分级方面,患者2年内复发率差异均无统计学意义.T3~4N0M0分期患者的短期预后与P53表达高低相关.结论:P53的过表达对晚期LSCC短期预后评估具有一定的临床意义,可考虑作为晚期LSCC分层治疗指标.     

Immunohistochemical analysis of p53, APE1, hMSH2 and ERCC1 proteins in actinic cheilitis and lip squamous cell carcinoma

Souza L R, Fonseca‐Silva T, Pereira C S, Santos E P, Lima L C, Carvalho H A, Gomez R S, Guimarães A L S & De Paula A M B
(2011) Histopathology 58 , 352–360
Immunohistochemical analysis of p53, APE1, hMSH2 and ERCC1 proteins in actinic cheilitis and lip squamous cell carcinoma Aims: This study has compared the tissue expression of the p53 tumour suppressor protein and DNA repair proteins APE1, hMSH2 and ERCC1 in normal, dysplastic and malignant lip epithelium. Methods and results:  Morphological analysis and immunohistochemistry were performed on archived specimens of normal lip mucosa (n = 15), actinic cheilitis (AC) (n = 30), and lip squamous cell carcinoma (LSCC) (n = 27). AC samples were classified morphologically according to the severity of epithelial dysplasia and risk of malignant transformation. LSCC samples were morphologically staged according to WHO and invasive front grading (IFG) criteria. Differences between groups and morphological stages were determined by bivariate statistical analysis. Progressive increases in the percentage of epithelial cells expressing p53 and APE1 were associated with increases in morphological malignancy from normal lip mucosa to LSCC. There was also a significant reduction in epithelial cells expressing hMSH2 and ERCC1 proteins in the AC and LSCC groups. A higher percentage of malignant cells expressing APE1 was found in samples with an aggressive morphological IFG grade. Conclusions:  Our data showed that epithelial cells from premalignant to malignant lip disease exhibited changes in the expression of p53, APE1, hMSH2 and ERCC1 proteins; these molecular change might contribute to lip carcinogenesis.     

喉癌组织中LKB1与VEGF -C表达的相关性研究

目的:检测喉癌组织中抑癌基因LKB1和血管内皮生长因子VEGF-C的表达情况,探讨VEGF-C在喉鳞状细胞癌、正常组织中的表达关系.方法:选取喉癌患者40例喉癌组织为实验组,40例癌旁正常组织(距肿瘤边缘＞5mm,且病理证实为正常组织)为对照组.采用逆转录聚合酶链反应(RT-PCR)技术分析两组之间LKB1mRNA和VEGF-CmRNA的表达情况.结果:LKB1在癌组织中阳性表达率低于癌旁组织(P＜0.05),VEGF-C在癌组织中阳性表达率高于癌旁组织(P＜0.05),均与组织分化程度有相关性(P＜0.05).LKB1与VEGF-C呈直线负相关.结论:在喉鳞状细胞癌发生的过程中,LKB1和VEGF -C发生异常表达,且与喉鳞状细胞癌的进展有关.     

喉鳞状细胞癌中OPN、MMP-9的表达及意义

目的研究骨桥蛋白(osteopontin,OPN)和MMP-9在喉鳞状细胞癌(laryngeal squamous cell carcinoma,LSCC)组织中的表达及与临床病理特征之间的相关性,分析两者在LSCC发生、发展过程中的机制。方法采用免疫组化SP法检测47例LSCC及10例正常喉咽黏膜中OPN和MMP-9的表达。结果 (1)47例LSCC组织中OPN、MMP-9蛋白的阳性率分别为63.8%、68.1%,在癌旁正常组织中阳性率为10%和0,两者比较差异有显著性(P<0.05);(2)OPN表达与LSCC临床分期、病理分级、淋巴结转移有关(P<0.05),MMP-9与LSCC临床分期、淋巴结转移有关(P<0.05),OPN和MMP-9的表达与患者年龄、性别、肿瘤位置无关(P>0.05);(3)LSCC组织中OPN和MMP-9的表达呈正相关(r=0.340,P=0.020)。结论 LSCC的发生、发展与OPN和MMP-9的高表达有关,OPN可能通过上调MMP-9的表达,降解细胞外基质,从而促进肿瘤的转移。     

Expression of cell cycle regulatory proteins in the multistep process of oesophageal carcinogenesis: stepwise over-expression of cyclin E and p53, reduction of p21(WAF1/CIP1) and dysregulation of cyclin D1 and p27(KIP1)

AIMS: Cell cycle regulatory proteins were analysed by immunohistochemistry in order to clarify how their expression changes with the degree of atypia as oesophageal surface squamous epithelium progresses from normal mucosa, through reactive change, low-grade dysplasia, and high-grade dysplasia to mucosal invasive carcinoma. METHODS AND RESULTS: Immunostaining for cyclin D1, cyclin E, p21, p27, p53 and Ki67 proteins was performed using 22 normal mucosa, 17 reactive change, 22 low-grade dysplasia, 15 high-grade dysplasia and 22 mucosal invasive carcinoma specimens. Normal mucosa, low-grade dysplasia and high-grade dysplasia samples were taken from patients without any oesophageal invasive carcinoma by endoscopic biopsy or endoscopic mucosal resection, and reactive change and mucosal invasive carcinoma were obtained from oesophagectomy material. Stepwise over-expression of cyclin E (P < 0.0001) and p53 (P < 0.0001), reduction of p21 (P=0.0189) and dysregulation of cyclin D1 and p27 were observed in the multistep process of oesophageal carcinogenesis. Significant differences in expression of p27 (P < 0.0001), p53 (P=0.0299) and Ki67 (P=0.0101) were observed between reactive change and low-grade dysplasia. Furthermore, expression of cyclin D1, cyclin E, p27 and p53 in mucosal invasive carcinoma were significantly different from those in high-grade dysplasia (P=0.0079, P=0.0237, P=0.0042 and P= 0.0299, respectively). CONCLUSIONS: Cell cycle regulatory proteins, cyclin E, p53 and p21 show stepwise over-expression or reduction with progression of oesophageal carcinogenesis, correlating with the increased cell proliferation observed with Ki67 labelling. We conclude that immunohistochemical analysis for p27, p53 and Ki67 is practically useful for the discrimination between low-grade dysplasia and reactive change. Cyclin D1, cyclin E, p27 and p53 help to distinguish high-grade dysplasia from mucosal invasive carcinoma.