魏茂富　卢义生　刘镜文　刘冠兰　张彩弟　康东平

[摘要] 目的：探讨p21、p27、bcl- 2蛋白在大肠癌、大肠腺瘤及正常大肠黏膜中的表达及其临床意义。方法：采用免疫组化方法,分别检测20例正常大肠黏膜、22例大肠腺瘤和69例大肠癌中p21、p27、bcl- 2蛋白表达情况。结果：p21、p27、bcl- 2蛋白总阳性表达率分别为44.93%、50.72%、52.17%，各种蛋白的阳性表达率在大肠癌组织与对照组间存在的差异有统计学意义(P<0.05) 。其中发生淋巴结转移的大肠癌组织中p21阳性表达率高于无淋巴结转移的大肠癌组织中的阳性表达率,差异具有统计学意义(P<0.05)；p27蛋白的表达与肿瘤分化程度呈负相关(P<0.05)；在发生淋巴结转移的大肠癌组织中bcl-2阳性表达率低于无淋巴结转移的大肠癌组织中的阳性表达率，差异具有统计学意义(P<0.05)，高分化、中分化腺癌与低分化腺癌相比，差异均有统计学意义(P<0.05)，TNM Ⅳ期bcl-2表达率显著低于Ⅰ、Ⅱ、Ⅲ期（P<0.05）。结论：p21、p27、bcl- 2蛋白与大肠癌的发生、发展有密切关系，联合检测p21、p27、bcl-2对于临床诊断大肠癌及判断病情、预后有重要意义。

[关键词] p21；p27；bcl-2；大肠腺瘤；大肠癌

[中图分类号]R735.3+4 [文献标识码]B [文章编号]1674-4721（2009）02（a）-022-04

Study of expression of p21, p27, bcl-2 in colorectal carcinoma, adenoma and normal tissues

WEI Mao-fu1,LU Yi-sheng2,LIU Jing-wen1,LIU Guan-lan1,ZHANG Cai-di1,KANG Dong-ping3

(1. Department of Pathology, Humen Hospital, Dongguan 523900,China;2. Department of Pathology, Peoplep's Hospital of Dongguan, Dongguan 523018,China;3.Department of Pathology, Taiping Hospital, Dongguan 523900,China)

[Abstract] Objective:To investigate the relationship between the expression of p21, p27, bcl-2 in the normal colorectal mucous membrane, colorectal adenoma ,carcinoma and clinicopathologic features. Methods:The expression of p21, p27, bcl-2 was detected respectively in 69 patients with colorectal carcinoma , 22 with colorectal adenoma and 20 normal colorectal mucous membrane by the immunohistochemical staining. Results:The frequency of p21, p27, bcl-2 staining in carcinoma tissues is 44.93%、50.72%、52.17%, respectively , being significantly difference in positive rates of these protein of control sample (P<0.05). The positive rate of p21 immunostaining was 70.83% in patients with lymphytic metastasis being significantly higher than that of in patients without lymphytic metastasis 31.11%(P<0.05) . A statistically significant difference in p27 staining was found in relation to the histologic grade (P<0.05). The positive rate of bcl-2 immunohistochemical staining was 33.33% in patients with lymphytic metastasis, compared with 62.22% in patients without lymphytic metastasis (P=0.022 1). A statistically significant difference in bcl-2 staining was found in relation to the histologic grade(P<0.05). Staining frequency for bcl-2 in TNM Ⅳ was significantly lower than those of TNM Ⅰ, Ⅱ, Ⅲ(P<0.05) . Conclusion:The results suggest that p21, p27, bcl-2 protein have relations with the genesis and development of colorectal carcinoma. The detection for these proteins will play an important part in clinical diagnosis and prognosis.

[Key words] p21; p27; bcl-2; Colorectal adenoma; Colorectal carcinoma

分子生物学研究表明大肠癌的发生、发展是一个多因素共同参与的多阶段复杂生物学过程, 其与癌基因激活, 抑癌基因失活密切相关。为此我们对大肠癌组织中p21、p27、bcl-2 基因同时进行检测, 以探讨各基因在大肠癌组织中表达的临床意义以及相互关系。

1材料与方法

1.1标本来源

收集我院及东莞市太平人民医院2003年3月～2008年3月手术切除经病理证实的大肠腺瘤标本22例,大肠癌标本69例，另外选取正常大肠黏膜标本20例作为对照观察。69例大肠癌标本中, 男38例,女31例，年龄28～82岁，平均年龄59.1岁,术前均未作任何抗肿瘤治疗。高分化腺癌5例,中分化腺癌17例,低分化腺癌47例。24例有淋巴结转移。TNM分期:Ⅰ期10例、Ⅱ期27例、Ⅲ期23例、Ⅳ期9例。

1.2试剂及免疫组化染色方法

鼠抗人p21、p27、bcl-2单克隆抗体、抗体稀释液及试剂盒均购自基因公司。p21稀释度为1∶50，P27、bcl-2稀释度为1∶200。所有标本均用10 %中性甲醛固定,石蜡包埋,4 μm 连续切片,按二步法法进行免疫组化染色,染色前标本切片进行抗原修复处理，一抗在室温孵育1 h，二抗在室温下孵育0.5 h，DAB 显色，苏木素复染。……