肝癌 Open Access
Copyright ©The Author(s) 2003. Published by Baishideng Publishing Group Inc. All rights reserved.
世界华人消化杂志. 2003-09-15; 11(9): 1362-1364
在线出版日期: 2003-09-15. doi: 10.11569/wcjd.v11.i9.1362
b-catenin和Cyclin D1在肝癌肝内转移中的作用
苏小康, 赵先明, 李锦清, 崔学教, 谢晓华, 杨海燕, 徐发彬, 石明
苏小康, 赵先明, 崔学教, 谢晓华, 杨海燕, 徐发彬, 广州中医药大学第一附属院三外科 广东省广州市 510405
苏小康, 男, 1969-07-25生, 陕西省商州市人, 汉族. 博士后, 讲师.
李锦清, 石明, 中山大学肿瘤医院肝胆科 广东省广州市 510060
通讯作者: 苏小康, 510405, 广东省广州市机场路16号, 广州中医药大学第一附属院三外科. suxibo@yeah.net
电话: 020-36591843 传真: 020-36590540
收稿日期: 2003-03-07
修回日期: 2003-03-17
接受日期: 2003-03-29
在线出版日期: 2003-09-15

目的

探讨b-catenin和Cyclin D1与癌栓形成及临床病理参数的关系.

方法

用免疫组化法检测32例肝癌原发灶、癌栓和癌旁组织中b-catenin、Cyclin D1的表达, 同时分析其与临床病理参数之间的关系.

结果

b-catenin在原发灶、癌栓和癌旁组织中的阳性率分别为62.5%、81.25%、9.38%, 三者之间有显著差别; b-catenin与癌栓形成、HBV感染、肝硬化有关. Cyclin D1在原发灶、癌栓和癌旁组织中的阳性率分别为46.9%、75%、18.8%, 三者之间有显著差别; Cyclin D1与癌栓形成、组织类型、分化程度有关.

结论

b-catenin和Cyclin D1促进癌栓形成, 参与HBV感染和肝硬化形成过程; Cyclin D1影响肿瘤组织类型和分化程度.

关键词: N/A
引文著录: 苏小康, 赵先明, 李锦清, 崔学教, 谢晓华, 杨海燕, 徐发彬, 石明. b-catenin和Cyclin D1在肝癌肝内转移中的作用. 世界华人消化杂志 2003; 11(9): 1362-1364
Role of b-catenin and cyclin D1 expressions in intrahepatic dissemination of liver cancer
Xiao-Kang Su, Xian-Ming Zhao, Jin-Qing Li, Xue-Jiao Cui, Xiao-Hua Xie, Hai-Yan Yang, Fa-Bing Xu, Ming Shi
Xiao-Kang Su, Xian-Ming Zhao, Xue-Jiao Cui, Xiao-Hua Xie, Hai-Yan Yang, Fa-Bing Xu, Third Department of General Surgery, First Affiliated Hospital, Guangzhou Traditional Chinese Medical University, Guangzhou 510405, Guangdong Provine, China
Jin-Qing Li, Ming Shi, Department of Hepatobiliary Surgery, Cancer Hospital, Zhongshan University, Guangzhou 510060, Guangzhou Province, China
Correspondence to: Xiao-Kang Su, Third Department of General Surgery, First Affiliated Hospital, Guangzhou Traditional Chinese Medical University, 16 Jichang Road, Guangzhou 510405, Guangzhou Province, China. Suxibo@yeah.net
Received: March 7, 2003
Revised: March 17, 2003
Accepted: March 29, 2003
Published online: September 15, 2003

AIM

To analyze the relationship between expressions of b-catenin and cyclin D1 and formation of portal vein tumor thrombus and clinical pathologic parameters.

METHODS

b-catenin and Cyclin D1 were tested in primary liver cancer, tumor thrombus and paracancer tissue in 32 cases of hepatic carcinoma with immunohistochemistry. Their effect on clinic pathologic parameters was also discussed.

RESULTS

The positive rates of b-catenin expression in primary liver cancer, tumor thrombus and paracancer tissue were 62.5%, 81.25%, and 9.38%, respectively. The positive rates of b-catenin expression were also correlated with tumor thrombus, HBV infection and liver cirrhosis. The positive rates of cyclin D1 expression in primary liver cancer, tumor thrombus and paracancer tissue were 46.9%, 75%, and 18.8%, respectively. It was also correlated with tumor thrombus, cell classification and differentiation. There were obvious differences among three groups in expression of b-catenin and cyclin D1.

CONCLUSION

b-catenin and cyclin D1 are helpful for initiation and progression of tumor thrombus and play an important role in the course of liver cirrhosis and HBV infection. Cyclin D1 influences tumor classification and differentiation.

Key Words: N/A
0 引言

b-catenin不仅在钙粘蛋白(E-CD)介导的细胞黏附、细胞分化、细胞骨架中起重要作用, 而且是WNT信号传导通道的关键环节, 调控Cyclin D1及C-myc等原癌因的表达. Cyclin D1能与细胞周期蛋白激酶(CDK4)结合, 激活的CDK4- Cyclin D1, 复合物可以与多种蛋白协同作用促进细胞由G1期向S期的过度, 我们检测了b-catenin和Cyclin D1蛋白在肝癌组织、门脉癌栓及癌旁组织中的表达, 探讨了b-catenin和Cyclin D1与肝癌肝内播散和临床病理参数之间的关系, 现报告如下

1 材料和方法
1.1 材料

2000-07/2001-05中山大学肿瘤医院肝胆外科手术切除原发性肝癌标本, 同时取肿瘤边缘2 cm外肝组织作癌旁组织, 男25例, 女7例, 年龄26-64, (中位45岁). 40 g/L甲醛固定, 常规石蜡包埋, 5 mm连续切片. 根据Enmondson病理标准分为I、II、III、IV 4级. b-catenin、Cyclin D1鼠抗人购自广州康润生物制品公司, SP试剂盒购自美国Vector公司.

1.2方法 免疫组化操作步骤按试剂盒说明操作, 并根据情况稍加改进, 两种方法均需微波抗原修复. 用试剂携带阳性片作为阳性对照, PBS液作为一抗作为阴性对照. 将阳性细胞按其数量及染色强度分为3级. 表达弱阳性(+): 细胞数量<10%, 染色强度弱阳性或仅个别细胞中至强阳性; 表达强阳性(+++): 阳性细胞数>60%, 中至强阳性染色, 少数可为弱阳性表达; 中度阳性(++): 阳性细胞数在10-60%之间; 无表达(-): Cyclin D1染色强度与背景基本无差别, b-catenin为细胞膜轻度染色.

统计学处理 用SPSS10.0行FISHER精确x2检验.

2 结果
2.1 b-catenin的表达

癌旁组织中b-catenin均匀分布于细胞膜, 仅极个别标本中有胞质极弱着色, 细胞核无染色, 而在原发灶62.5% (20/32), 癌栓81.25% (26/32)中出现在细胞质或核内沉积. 门静脉癌栓明显高于原发灶(P <0.04), 而在无门脉癌栓者原发灶中表达强度明显低于有转移灶者 (表1).

表1 HCC、癌栓、癌旁组织中b-catenin和cyclin D1蛋白的表达(n =32).
HCCb-catenin
cyclin D1
-++++++-++++++
原发灶1297417654
癌栓669 a1184812a
癌旁2921026420
aP <0.05.
2.2 Cyclin D1的表达

Cyclin D1阳性染色有二种类型; 细胞核型: 定位于细胞核中, 细胞质型: 分布于细胞质中, 阳性细胞染色呈弥漫状分布. 肝癌原发灶中46.9%呈阳性表达, 癌栓中阳性率则高达75.0%, 而癌旁组织中明显低, 仅18.8%, 在三组间有明显差异, 依次为癌栓、原发性、癌旁 (表1) .

2.3 b-Catenin, Cyclin D1与临床病理参数之间的关系

b-Catenin的异常表达与HBV感染、肝硬化和癌栓形成有关, 而与肿瘤大小、包膜、组职类型和分化程度无关. Cyclin D1在癌栓形成、组织类型和分化中有差异, 而在HBV感染, 肝硬化, 肿瘤大小、包膜中无差异(表2).

表2 b-catenin、cyclin D1与临床病理参数之间的关系.
参数b-catenin
cyclin D1
NANA
HBV+714109
-56a76
肝硬化轻71699
重54a86
肿瘤大小 小于5 cm6121010
大于5 cm6875
包膜完整71198
不完整5987
癌栓有315413
无95a132a
组织类型HCC69510
CC611125a
混合型1100
分化程度I4646
II61097a
III2442
N: 细胞核 A: 细胞质
aP <0.05
3 讨论

门静脉癌栓是原发性肝癌肝内转移的主要方式, 也是引起肝癌术后复发的重要原因(术后复发率高达60.8%), 严重影响临床疗效[1,2]. b-Catenin在细胞黏附和WNT信号传导两个过程发挥作用, c-myc和Cyclin D1是WNT通路中的核心环节[3,4], 无功能的b-Catenin在细胞过度沉积导致细胞内游离b-Catenin水平升高, 细胞过度增生[5,6]. 我们发现在转移灶中有b-Catenin异常表达, 而且在癌栓中明显高于原发灶和癌旁组织, 参与癌栓形成过程中多个环节: 黏附松散, 脱离复发灶, 到达转移部位的增生和黏附两方面发挥作用, 通过两个途径促进肝内播散. Cyclin D1编码基因位于11q3[7,8], 我们发现原发灶, 癌栓和癌变组织中表达不同, 癌栓中明显高于原发灶和癌旁组织. 提示Cyclin D1在原发灶形成和肿瘤细胞转移到门脉内继续增生都有促进作用.

b-Catenin作为WNT通路中的关键环节, 并非单一上下游的线性运行[9-11], 与其他信号通道可能相互影响, 从而形成复杂的网络效应, Gleason et al [12]人发现 WNT-1, Notch信号通路G蛋白通路之间有相互作用, 提示b-Catenin异常表达在肿瘤发展过程中有多重效应[13-15]. 本研究发现HBV感染肝硬化以及门脉癌栓形成与b-Catenin的异常表达密不可分, 可能在HBV感染或HBX基因插入诱导肝细胞恶变过程, 肝脏损伤纤维化修复过程中有b-Catenin或其信号传导机制的异常. 但是本研究发现b-Catenin对肿瘤大小, 包膜, 组织类型及分化程度的影响不大. Cyclin D1是一种弱致癌基因, 与其他癌基因关系密切[16-18], 在一些细胞种Cyclin D1本身并无转化作用[19-21]. 但能与ras基因协同转化大鼠成纤维细胞[22-24]. 野生型 P53蛋白的积聚可引起Cyclin D1表达增加, Cyclin D1基因扩增和P53突变是正相关, 因而cyclin D1异常表达进一步影响肿瘤临床病理参数[25,26], Cyclin D1表达与乳腺癌组织类型有关[27]. 在胰腺癌研究中, 发现Cyclin D1表达与年龄、分化程度、分期大小、部位、淋巴结转移无关, 但Cyclin D1阴性者生存期明显短[28,29], 本组资料中组织类型、分化程度、癌栓形成与Cyclin D1异常表达有关, 可能是Cyclin D1与其他癌基因协同作用的结果. Qin et al [30]也发现肝癌中有Cyclin D1基因扩增或过度表达, 而且影响临床病理参数.

本结果显示b-Catenin和Cyclin D1蛋白表达与肝癌肝内播散有关, 促进门静脉癌栓形成, 与部分临床病理参数密切相关, 通过WNT信号通路及细胞周期途径影响肝癌的发展, 针对上述环节的分子生物学治疗可能会阻止门脉癌栓的形成, 可以作为开展分子生物学治疗的靶点. 但体内信号传递网络的复杂性和细胞周期调控的精确机制不是b-Catenin和Cyclin D1能完全解释的, 对信号传递和细胞周期调控的进一步研究可能会给肝癌转移的治疗带来突破性的进展.

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