CyclinD1、p27和Ki-67在细支气管肺泡癌中的表达及其意义
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摘要
细支气管肺泡癌（简称BAC）是发生于细支气管Clara细胞和Ⅱ型
肺泡细胞的一种特殊型肺腺癌，近年来其发病率呈逐渐增加趋势。BAC
的病因及发病机理不明，近来在癌基因及抑癌基因方面的研究大多是以
包括BAC在内的腺癌或非小细胞肺癌为主体进行的，国内外对BAC单
独进行研究很少。
近年研究表明，细胞周期失控是细胞增殖失控而导致癌变的重要原
因。参与细胞周期调控的分子有细胞周期蛋白（cyclin）、细胞周期蛋白
依赖性激酶（CDK）和CDK抑制蛋白（CDKI）。细胞周期的不同时相由
不同的调节因子控制，其中G_1期的调节因子与癌变的关系最为密切。Cyclin
D1基因是近年来提出的重要的原癌基因，其编码的cyclin D1蛋白在细胞
周期G_1/S的转换中发挥重要的正性调控作用，与肿瘤的发生密切相关，
在人类多种恶性肿瘤中发现有cyclin D1蛋白的过表达。p27是近年来发
现的一个重要的CDKI，主要通过抑制G_1期的cyclin-CDK的活性，使细
胞不能通过G_1期。有研究表明p27表达下降与多种肿瘤的发生和发展有
关，并且可以作为一个独立的预后指标。目前关于BAC中cyclin D1表
达的研究报道很少，p27在BAC中的表达尚未见报道。Ki-67是一种细胞
周期相关抗原，是目前应用最广泛的增殖细胞标记之一。目前关于Ki-67
的表达与临床病理参数之间的关系在不同的肿瘤中研究结果各不相同。
本文采用免疫组化方法对我院手术切除的43例BAC标本进行cyclin D1、
昨7和K卜67表达水平检测，并探讨它们之间的关系及临床病理学意义。
材料与方法
43例BACS取自浙江大学医学院附属邵逸夫医院1994－2000年间手术切除的
标本，另选13例癌旁正常肺组织作为对照组。按洲0（1999）的标准进行组织
学分型，非粘液型39例，粘液型4例。按国际统一的TNM分期法进行临床分期，
互期 19例，11期 12例，Ill期 6例，IV期 6例。采川免疫组化En Vision二步
法检测cyclh DI、p27利】K卜67的表达，一抗卜作浓度均为1：SO，设立
叫性和阴性对照。川性判断和分级标准：三者均以细胞核Y棕黄色为阳
性细胞，据p［I性细胞所11；十b例并适当参考川性强度进行半定量分级：＜5％（－），
50－25O（＋），25O－50O（＋＋），＞500（＋＋＋）。统计学处理采用X’检验和确切
概率法。
结果与讨论
叼例＊＊O中叮卜nDI川性表达者28例，川性率65．1％，而门例
止常肺组织未见。yclln DI川性表达，两者具非常显岔差异（确切概率法，
p＜0．0！）。此结果表明 CyClifl DI过表达与 BAC的发生密切相大。有淋巳
结转移组中原发癌利淋巴结转移癌cyclh DI的1川性率相同。cyclln DI的
表达与患者的性别、年龄、吸烟与否、绢织学类刑、淋巴结转移状况和
临床期别均无关，但与肿瘤大小有关，＜3cffi组eye！h DI 的川性率明显
高于）3cm组，差异有显著性（确切概率法，p＜0刀5）。BAC中cyclin DI
的表达与肿瘤大小呈负相关，此结果与文献报道不一致。我们认为这一
结果由卜列原回引起：（l）肿瘤是多基因损伤的结果，极可能在部分BAC
中存在更重要的抑癌基冈或癌基囚发挥了作用；（2）文献报道当 cyclin DI
的表达超过某个极限时即可能产生细胞毒作用，反而可抑制增殖。
43例 BACS中 p27 pl－I性表达者 22例，阳性率 sl。2O，明显低于上常
肺组织的92．3％，两者具非常显著差异（确切概率法，p功刀1）。有淋巴
结转移组中原发癌利淋巴结转移癌p27 的阳性率相近。与临床病理参数
2
之间的分析显示：p27的表达与患者的性别、年龄、吸烟与否、肿瘤大小、
组织学类型均无关；有淋巴结转移组p27的阳性率明显低于无淋巴结转
移组，差异有显著性（X‘检验，p＜0．05）；临床Ill＋IV期p27的阳性率明显
低于卜11期，差异有显著性（确切概率法，p＜0刀5）。上述结果表明，p27
蛋白表达下降与BAC的发生和发展有关，提示其为预后不良的标志。对
BAC患者，检测p27的表达水平将有助于判断预后及指导治疗。
43例BACS中M－67 F厂牲表达者23例，川性率53．5％，较正常肺组
织显芦升高，后者在支气管上皮中偶见散在的川性核，两者差异非常显
符（确切概率法，p＜0刀 1）。有淋巴结转移组中原发癌和淋巴结转移癌 Ki－
67 的m性率无显斡差异。目前关于Ki67 的表达与临床病理参数间的关
系在不同的肿瘤中结果不同，本组结果显示Ki67的表达与各临床病理参
数均无关，其中有淋巴结转移组K卜67的川性率（63．＊％）高丁无淋巴结
转移组（叱刀％〕，但斧异不显著（／检验，p叫．门2〕。关于8＊C中K卜盯
表达的确切意义，尚需扩人病例数进一步研究证实。
关丁叮1山DI、咋7和K广67 i白表达间的大系，文献报道在不同的肿瘤
中结果不同，本实验结果显示BAC中 cyclin DI、p27和 Ki－67 i白表达之间无
相关性。
结论
1．BAC中cvclin DI的表达显著增强，表达水平与肿瘤人小呈
Bronchioloalveolar carcinoma(BAC) is a particular subtype of pulmanary adenocarcinoma,which derives from clara cell and type II pneumocyte. The incidence of BAC appears to be rising recently. The etiology and pathogenesis of this unique neoplastic disease are still unclear, many studies of oncogene and tumor suppressor gene expression include BAC with all adenocarcinoma or non-small cell lung cancer, rather than as a separate category.
Recent studies have revealed that uncontrolled cell cycle is very important to uncontrolled cell proliferation and carcinogenesis. Cyclin-dependent kinase (CDK), together with cyclin and CDK inhibitor, govern cell cycle progression in eukaryotic cell. In cell cycle, different phase is regulated by different factors, in them, Grphase regulators are most closely related to carcinogenesis. Proto-oncogene cyclin Dl encodes cyclin Dl protein, which acts as an important positive regulator in G1/S transition, and appears to be closely linked with carcinogenesis, the overexpression of cyclin Dl has been reported in a variety of malignant tumors. p27 is an important CDK inhibitor, which binds to and inhibits preferentially G1-phase kinases thereby arresting the cell cycle at G1-phase. Decreased p27 expression has been shown to associated with the development and malignant progression in many human malignant tumors, and
provides independent prognostic information. To our knowledge, p27 expression in BAC has not previously been reported in the literature, cyclin Dl expression in BAC not reported except for 19 cases in the study of McDonald et al. Ki-67 antigen is a cell cycle-related nuclear antigen, anti-Ki67 antibody is widely used for detecting proliferating cells. The relationship between Ki-67 expression and clinicopathological parameters is different in different tumors. Here we evaluated the expression of the cell cycle-related proteins cyclin Dl, p27 and Ki-67 in 43 surgically resected BACs using immunohistochemical En Vision method, discussed the relationship between the expression of the genes and their clinicopathological significances.
Materials and Methods
43 cases BACs and 13 paracancerous normal pulmonary tissue were taken from the surgically resected specimen in Sir Run Run Shaw Hospital from 1994 to 2000. Histological subtyping of BAC according to WHO(1999) criteria revealed that 39 were nonmucinous, 4 were mucinous. According to the international TNM staging system, 19 patients were classfied at stage I, 12 at stage II, 6 at stage III, 6 at stage IV. Expression of cyclin Dl, p27 and Ki-67 was detected by immunohistochemical En Vision method, with 1:50 dilution of primary antibody of cyclin Dl, p27 and Ki-67, positive and negative controls were designed. Judging and grading system: for each antibody, positive immunostaining appears as yellow brown nucleus with clear background, the immunohistochemical stains were graded on a semiquantitative scale according to the prevalence of nuclear staining and combined with staining color intensity: <5% positive cells (-), 5%~25%(+), 25%~50%(++), >50%(+++). Statistical analysis: chi-square test and Fisher' s exact test.
Results and Discussion
Cyclin Dl expression was seen in 28/43 BACS(65. 1%), while no expression was observed in matched pulmonary tissue, the difference was
very significant(Fisher's exact test, p<0.01). This result indicates that cyclin Dl overexpression plays an important role in the development of BAC. In the group with lymph node metastasis, the primary tumor and metastatic tumor showed the same positivity. No statistically significant association was found between cyclin Dl expression data and sex, age, tobacco use history, subtype of BAC(mucinous vs. nonmucinous), lymph node status and clinical stage, but the cyclin Dl expression was negatively correlated with tumor size, the positivity in <3cm group was significantly higher than in >3cm group (Fisher' s exact test, p<0. 05), this finding is not in agreement with literature observations. We think this result may be caused by the following rea
肺泡细胞的一种特殊型肺腺癌，近年来其发病率呈逐渐增加趋势。BAC
的病因及发病机理不明，近来在癌基因及抑癌基因方面的研究大多是以
包括BAC在内的腺癌或非小细胞肺癌为主体进行的，国内外对BAC单
独进行研究很少。
近年研究表明，细胞周期失控是细胞增殖失控而导致癌变的重要原
因。参与细胞周期调控的分子有细胞周期蛋白（cyclin）、细胞周期蛋白
依赖性激酶（CDK）和CDK抑制蛋白（CDKI）。细胞周期的不同时相由
不同的调节因子控制，其中G_1期的调节因子与癌变的关系最为密切。Cyclin
D1基因是近年来提出的重要的原癌基因，其编码的cyclin D1蛋白在细胞
周期G_1/S的转换中发挥重要的正性调控作用，与肿瘤的发生密切相关，
在人类多种恶性肿瘤中发现有cyclin D1蛋白的过表达。p27是近年来发
现的一个重要的CDKI，主要通过抑制G_1期的cyclin-CDK的活性，使细
胞不能通过G_1期。有研究表明p27表达下降与多种肿瘤的发生和发展有
关，并且可以作为一个独立的预后指标。目前关于BAC中cyclin D1表
达的研究报道很少，p27在BAC中的表达尚未见报道。Ki-67是一种细胞
周期相关抗原，是目前应用最广泛的增殖细胞标记之一。目前关于Ki-67
的表达与临床病理参数之间的关系在不同的肿瘤中研究结果各不相同。
本文采用免疫组化方法对我院手术切除的43例BAC标本进行cyclin D1、
昨7和K卜67表达水平检测，并探讨它们之间的关系及临床病理学意义。
材料与方法
43例BACS取自浙江大学医学院附属邵逸夫医院1994－2000年间手术切除的
标本，另选13例癌旁正常肺组织作为对照组。按洲0（1999）的标准进行组织
学分型，非粘液型39例，粘液型4例。按国际统一的TNM分期法进行临床分期，
互期 19例，11期 12例，Ill期 6例，IV期 6例。采川免疫组化En Vision二步
法检测cyclh DI、p27利】K卜67的表达，一抗卜作浓度均为1：SO，设立
叫性和阴性对照。川性判断和分级标准：三者均以细胞核Y棕黄色为阳
性细胞，据p［I性细胞所11；十b例并适当参考川性强度进行半定量分级：＜5％（－），
50－25O（＋），25O－50O（＋＋），＞500（＋＋＋）。统计学处理采用X’检验和确切
概率法。
结果与讨论
叼例＊＊O中叮卜nDI川性表达者28例，川性率65．1％，而门例
止常肺组织未见。yclln DI川性表达，两者具非常显岔差异（确切概率法，
p＜0．0！）。此结果表明 CyClifl DI过表达与 BAC的发生密切相大。有淋巳
结转移组中原发癌利淋巴结转移癌cyclh DI的1川性率相同。cyclln DI的
表达与患者的性别、年龄、吸烟与否、绢织学类刑、淋巴结转移状况和
临床期别均无关，但与肿瘤大小有关，＜3cffi组eye！h DI 的川性率明显
高于）3cm组，差异有显著性（确切概率法，p＜0刀5）。BAC中cyclin DI
的表达与肿瘤大小呈负相关，此结果与文献报道不一致。我们认为这一
结果由卜列原回引起：（l）肿瘤是多基因损伤的结果，极可能在部分BAC
中存在更重要的抑癌基冈或癌基囚发挥了作用；（2）文献报道当 cyclin DI
的表达超过某个极限时即可能产生细胞毒作用，反而可抑制增殖。
43例 BACS中 p27 pl－I性表达者 22例，阳性率 sl。2O，明显低于上常
肺组织的92．3％，两者具非常显著差异（确切概率法，p功刀1）。有淋巴
结转移组中原发癌利淋巴结转移癌p27 的阳性率相近。与临床病理参数
2
之间的分析显示：p27的表达与患者的性别、年龄、吸烟与否、肿瘤大小、
组织学类型均无关；有淋巴结转移组p27的阳性率明显低于无淋巴结转
移组，差异有显著性（X‘检验，p＜0．05）；临床Ill＋IV期p27的阳性率明显
低于卜11期，差异有显著性（确切概率法，p＜0刀5）。上述结果表明，p27
蛋白表达下降与BAC的发生和发展有关，提示其为预后不良的标志。对
BAC患者，检测p27的表达水平将有助于判断预后及指导治疗。
43例BACS中M－67 F厂牲表达者23例，川性率53．5％，较正常肺组
织显芦升高，后者在支气管上皮中偶见散在的川性核，两者差异非常显
符（确切概率法，p＜0刀 1）。有淋巴结转移组中原发癌和淋巴结转移癌 Ki－
67 的m性率无显斡差异。目前关于Ki67 的表达与临床病理参数间的关
系在不同的肿瘤中结果不同，本组结果显示Ki67的表达与各临床病理参
数均无关，其中有淋巴结转移组K卜67的川性率（63．＊％）高丁无淋巴结
转移组（叱刀％〕，但斧异不显著（／检验，p叫．门2〕。关于8＊C中K卜盯
表达的确切意义，尚需扩人病例数进一步研究证实。
关丁叮1山DI、咋7和K广67 i白表达间的大系，文献报道在不同的肿瘤
中结果不同，本实验结果显示BAC中 cyclin DI、p27和 Ki－67 i白表达之间无
相关性。
结论
1．BAC中cvclin DI的表达显著增强，表达水平与肿瘤人小呈
Bronchioloalveolar carcinoma(BAC) is a particular subtype of pulmanary adenocarcinoma,which derives from clara cell and type II pneumocyte. The incidence of BAC appears to be rising recently. The etiology and pathogenesis of this unique neoplastic disease are still unclear, many studies of oncogene and tumor suppressor gene expression include BAC with all adenocarcinoma or non-small cell lung cancer, rather than as a separate category.
Recent studies have revealed that uncontrolled cell cycle is very important to uncontrolled cell proliferation and carcinogenesis. Cyclin-dependent kinase (CDK), together with cyclin and CDK inhibitor, govern cell cycle progression in eukaryotic cell. In cell cycle, different phase is regulated by different factors, in them, Grphase regulators are most closely related to carcinogenesis. Proto-oncogene cyclin Dl encodes cyclin Dl protein, which acts as an important positive regulator in G1/S transition, and appears to be closely linked with carcinogenesis, the overexpression of cyclin Dl has been reported in a variety of malignant tumors. p27 is an important CDK inhibitor, which binds to and inhibits preferentially G1-phase kinases thereby arresting the cell cycle at G1-phase. Decreased p27 expression has been shown to associated with the development and malignant progression in many human malignant tumors, and
provides independent prognostic information. To our knowledge, p27 expression in BAC has not previously been reported in the literature, cyclin Dl expression in BAC not reported except for 19 cases in the study of McDonald et al. Ki-67 antigen is a cell cycle-related nuclear antigen, anti-Ki67 antibody is widely used for detecting proliferating cells. The relationship between Ki-67 expression and clinicopathological parameters is different in different tumors. Here we evaluated the expression of the cell cycle-related proteins cyclin Dl, p27 and Ki-67 in 43 surgically resected BACs using immunohistochemical En Vision method, discussed the relationship between the expression of the genes and their clinicopathological significances.
Materials and Methods
43 cases BACs and 13 paracancerous normal pulmonary tissue were taken from the surgically resected specimen in Sir Run Run Shaw Hospital from 1994 to 2000. Histological subtyping of BAC according to WHO(1999) criteria revealed that 39 were nonmucinous, 4 were mucinous. According to the international TNM staging system, 19 patients were classfied at stage I, 12 at stage II, 6 at stage III, 6 at stage IV. Expression of cyclin Dl, p27 and Ki-67 was detected by immunohistochemical En Vision method, with 1:50 dilution of primary antibody of cyclin Dl, p27 and Ki-67, positive and negative controls were designed. Judging and grading system: for each antibody, positive immunostaining appears as yellow brown nucleus with clear background, the immunohistochemical stains were graded on a semiquantitative scale according to the prevalence of nuclear staining and combined with staining color intensity: <5% positive cells (-), 5%~25%(+), 25%~50%(++), >50%(+++). Statistical analysis: chi-square test and Fisher' s exact test.
Results and Discussion
Cyclin Dl expression was seen in 28/43 BACS(65. 1%), while no expression was observed in matched pulmonary tissue, the difference was
very significant(Fisher's exact test, p<0.01). This result indicates that cyclin Dl overexpression plays an important role in the development of BAC. In the group with lymph node metastasis, the primary tumor and metastatic tumor showed the same positivity. No statistically significant association was found between cyclin Dl expression data and sex, age, tobacco use history, subtype of BAC(mucinous vs. nonmucinous), lymph node status and clinical stage, but the cyclin Dl expression was negatively correlated with tumor size, the positivity in <3cm group was significantly higher than in >3cm group (Fisher' s exact test, p<0. 05), this finding is not in agreement with literature observations. We think this result may be caused by the following rea
引文
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18．姜彦多，王继红，夏书月．肺癌组织中细胞周期蛋白D1的检测及其意义，中国实验诊断学，1999；3(2)：67-69
19．胡向阳，饶慧蓉．Cyclin D1、Rb基因蛋白在乳腺癌中的表达．临床与实验病理学杂志，1998；14(6)：558-560
20.Bartkova J, Luks J, Muller H, et al. Cyclin D1 protein expression and function in human breast cancer. Int J cancer, 1994; 57:353-361
21.Ponce-Castaneda MV, Lee MH, Latres E, et al.p27kipl:chromosomal mapping to 12p12-12p13. 1 and absence of mutations in human tumors. CancerRes, 1995; 55:1211-1214
22.Kawamata N, Morrosetti R, Miler CW, et al. Molecular analysis of the cyclindependent kinase inhibitor gene p27/kipl in human malignancies. Cancer Res,1995; 55:2266-2269
23.Fero ML, Rivkin M, Tasch M, et al. A syndrome ofmultiorgan hyperplasia with features of gigantism, tumorigenesis, and female sterility in p27(kip 1)-deficient mice. Cell, 1996;85:733- 744
24.Tan P, Cady B,Wanner M, et al. The cell cycle inhibitor p27kipl is an independent prognostic marker in small (Tla, b) invasive breast carcinomas. Cancer Res, 1997;57:1259-1263
25.Tsihlias J, Kapusta LR, DeBoer G, et al. Loss of cyclin-dependent kinase inhibitor p27kipl is a novel prognostic factor in localized human prostate adenocarcinoma. Cancer Res, 1998; 1998; 58:542-548
26.Chiarle R, Budel LM, Skolnik J, et al. Increased proteasome degradation of cyclindependent kinase inhibitor p27 is associated with a decreased overall survival in mantle cell lymphoma. Blood, 2000; 95(2): 619-626
27．王华，王靖华，步宏等．p27kipl在子宫内膜癌中的表达及其意义．临床与实验病理学杂志，2000；16(4)：265-268
28．王福生，蒋丹斌，宋怀宇等．p27kipl在胃癌中的表达及与临床病理分期和预后的关系．临床与实验病理学杂志，2000；16(6)：487-488
29.Ito Y, Takeda T, Sakon M, et al.Expression and clinical significance of the G_1-S modulators in carcinoma of the extrahepatic bile duct. Anticancer Res, 2000; 20(lA): 337-344
30.Tut VM, Braithwaite KL, Angus B, et al. Cyclin D1 expression in transitional cell carcinoma of the bladder:cirrelation with p53, wafl, pRb and Ki-67. Br J Cancer, 2001;84(2): 270-275
31.Basolo F, Caligo MA, Pinchera a, et al. Cyclin D1 overexpression in thyroid carcinomas: relation with clinico-pathological parameters,retinoblastoma gene product and Ki-67 labelling index. Thyroid, 2000; 10(9): 741-746
32.Kim H J, Jung WH, Kim DY, et al. Expression of cyclins in ductal hyperplasia, atypical ductal hyperplasis and ductal carcinoma in situ of the breast. Yonsei Med,2000; 41 (3): 345-353
33.Dosaka—Akita H, Hommura F, Mishina T, et al. A risk stratification model of nonsmall cell lung cancer using Cyclin E, Ki-67 and ras p21: different roles of G_1 cyclins in cell proliferation and prognosis. Cancer, 2001; 61(6): 2500-2504
34.Hayashi H, Ito T, Yazawa T, et al. Reduced expression ofp27kipl is associated
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35． Hui AM, Cui X, Makuuchi M,et al. Decreased p27(kipl)expression and Cyclin D1 overexpression, alone and in combinition, influence recurrence and survival of pa-tients with resectable extrahepatic bile duct carcinoma. Hepatology, 1999; 30(5) : 1167-1173
36． Nohara T, Ryo T, Iwamoto S, et al. Expression of cell-cycle regulator p27 is correlated to the prognosis and ER expression in breast carcinoma patients. Oncology, 2001; 60(1) : 94-100
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16.Nguyen VN, Mirejovsky P, Mirejovsky T, et al. Expression of Cyclin D1, Ki-67 and PCNA in non-small cell lung cancer: prognostic significance and comparison with p53 and bcl-2. Acta Histochem, 2000; 102(3): 328-338
17．李本全，李清泉，潘显光．细胞周期蛋白D1在肺癌中的表达及价值．中华结核和呼吸杂志，1999；(22)5：316
18．姜彦多，王继红，夏书月．肺癌组织中细胞周期蛋白D1的检测及其意义，中国实验诊断学，1999；3(2)：67-69
19．胡向阳，饶慧蓉．Cyclin D1、Rb基因蛋白在乳腺癌中的表达．临床与实验病理学杂志，1998；14(6)：558-560
20.Bartkova J, Luks J, Muller H, et al. Cyclin D1 protein expression and function in human breast cancer. Int J cancer, 1994; 57:353-361
21.Ponce-Castaneda MV, Lee MH, Latres E, et al.p27kipl:chromosomal mapping to 12p12-12p13. 1 and absence of mutations in human tumors. CancerRes, 1995; 55:1211-1214
22.Kawamata N, Morrosetti R, Miler CW, et al. Molecular analysis of the cyclindependent kinase inhibitor gene p27/kipl in human malignancies. Cancer Res,1995; 55:2266-2269
23.Fero ML, Rivkin M, Tasch M, et al. A syndrome ofmultiorgan hyperplasia with features of gigantism, tumorigenesis, and female sterility in p27(kip 1)-deficient mice. Cell, 1996;85:733- 744
24.Tan P, Cady B,Wanner M, et al. The cell cycle inhibitor p27kipl is an independent prognostic marker in small (Tla, b) invasive breast carcinomas. Cancer Res, 1997;57:1259-1263
25.Tsihlias J, Kapusta LR, DeBoer G, et al. Loss of cyclin-dependent kinase inhibitor p27kipl is a novel prognostic factor in localized human prostate adenocarcinoma. Cancer Res, 1998; 1998; 58:542-548
26.Chiarle R, Budel LM, Skolnik J, et al. Increased proteasome degradation of cyclindependent kinase inhibitor p27 is associated with a decreased overall survival in mantle cell lymphoma. Blood, 2000; 95(2): 619-626
27．王华，王靖华，步宏等．p27kipl在子宫内膜癌中的表达及其意义．临床与实验病理学杂志，2000；16(4)：265-268
28．王福生，蒋丹斌，宋怀宇等．p27kipl在胃癌中的表达及与临床病理分期和预后的关系．临床与实验病理学杂志，2000；16(6)：487-488
29.Ito Y, Takeda T, Sakon M, et al.Expression and clinical significance of the G_1-S modulators in carcinoma of the extrahepatic bile duct. Anticancer Res, 2000; 20(lA): 337-344
30.Tut VM, Braithwaite KL, Angus B, et al. Cyclin D1 expression in transitional cell carcinoma of the bladder:cirrelation with p53, wafl, pRb and Ki-67. Br J Cancer, 2001;84(2): 270-275
31.Basolo F, Caligo MA, Pinchera a, et al. Cyclin D1 overexpression in thyroid carcinomas: relation with clinico-pathological parameters,retinoblastoma gene product and Ki-67 labelling index. Thyroid, 2000; 10(9): 741-746
32.Kim H J, Jung WH, Kim DY, et al. Expression of cyclins in ductal hyperplasia, atypical ductal hyperplasis and ductal carcinoma in situ of the breast. Yonsei Med,2000; 41 (3): 345-353
33.Dosaka—Akita H, Hommura F, Mishina T, et al. A risk stratification model of nonsmall cell lung cancer using Cyclin E, Ki-67 and ras p21: different roles of G_1 cyclins in cell proliferation and prognosis. Cancer, 2001; 61(6): 2500-2504
34.Hayashi H, Ito T, Yazawa T, et al. Reduced expression ofp27kipl is associated
with the development of pulmonary adenocarcinoma.J Pathol, 2000; 192(1) : 26-31
35． Hui AM, Cui X, Makuuchi M,et al. Decreased p27(kipl)expression and Cyclin D1 overexpression, alone and in combinition, influence recurrence and survival of pa-tients with resectable extrahepatic bile duct carcinoma. Hepatology, 1999; 30(5) : 1167-1173
36． Nohara T, Ryo T, Iwamoto S, et al. Expression of cell-cycle regulator p27 is correlated to the prognosis and ER expression in breast carcinoma patients. Oncology, 2001; 60(1) : 94-100