【摘要】 背景:细胞周期调控蛋白水平及其相互关系是良恶性细胞增殖的主要区别标准。目的:了解肺癌组织p21waf1和p53及增殖细胞核抗原蛋白表达情况。设计:病例对照。单位:武汉大学人民医院呼吸内科。对象:来自1996-10/1999-05武汉大学人民医院保存的胸外科手术和纤维支气管镜活检组织石蜡包埋标本135块。其中肺癌76块,肺良性疾病59块。肺癌组中患者男56例,女20例;年龄18～74岁。肺良性疾病组中患者男42例,女17例;年龄16～70岁。方法:用磷酸盐缓冲液代替一抗作为阴性对照。染色步骤按试剂盒说明进行。p21waf1采用高温高压修复法,p53采用微波修复法,增殖细胞核抗原无需抗原修复。①p21waf1和p53阳性染色判断标准:计数连续5个高倍镜视野中胞核染成黄褐色的阳性细胞所占比例,以≥10%为阳性,<10%为阴性。②增殖细胞核抗原阳性判断标准:连续观察5个高倍镜视野,每个高倍镜视野计数100个细胞中胞核染成黄褐色的阳性细胞所占比例,取其平均值,记为增殖细胞核抗原标记指数。主要观察指标:p21waf1和p53及增殖细胞核抗原蛋白表达水平。结果:①肺癌组织p21waf1和p53蛋白表达阳性率分别为75%(57/76)和47%(36/76),增殖细胞核抗原标记指数显著高于肺良性疾病组织(0.44±0.32,0.09±0.14)。②肺癌组织p21waf1和p53及增殖细胞核抗原蛋白表达与肺癌临床分期及细胞分化程度无关。③肺鳞癌增殖细胞核抗原标记指数明显高于小细胞肺癌(0.51±0.33,0.30±0.36)。④肺癌组织p21waf1和p53表达与增殖细胞核抗原无关,而肺良性疾病组织p21waf1表达与增殖细胞核抗原有相关性。结论:肺癌组织p21waf1与p53蛋白表达明显上调,肺癌细胞增殖程度高。肺鳞癌细胞DNA损伤修复能力可能高于小细胞肺癌。肺良性疾病细胞增殖过程中,p21waf1和p53及增殖细胞核抗原3种蛋白作用协调,而肺癌细胞中3种蛋白作用不协调。更多还原

【Abstract】 BACKGROUND: The different level of proteins regulating cell cycle and their correlation is the main criteria to differentiate the benign and malignant cellular proliferation.OBJECTIVE: To investigate the expression status of p21waf1 and p53 in lung cancer as well as proliferating cell nuclear antigen (PCNA)DESIGN: A case-control study.SETTING: Department of Respiratory Medicine, Renmin Hospital , Wuhan UniversityPARTICIPANTS: This case-control study involved 135 patients who underwent lobectomy or fiberoptic bronchoscopy for primary lung cancer or benign chronic pulmonary diseases at Renmin Hospital of Wuhan University from October 1996 through May 1999. They were divided into two groups: lung cancer group (76 patients, including 56 men and 20 women, aged 18-74 years of age) and chronic pulmonary diseases group (59 cases,including 42 men and 17 women, aged 16-70 years of age).METHODS: Phosphate buffer solution replaced the first antibody as the negative control. Immunohistochemistry was performed using a modified streptavidin-biotinylated peroxidase technique according to the manufacturer’s recommendations (Maxim Corporation). For p21waf1 staining, we used hydrated autoclaving as a pretreatment. Antigen retrieval was performed in a standard microwave unit for p53 staining. PCNA staining did not need antigen retrieval.① Estimation criteria of p21waf1and p53positive staining: The ratio of the positive cells indicated by yellowish brown nucleus due to staining was counted for 5 successive high-fold microscopic fields: when it was≥10%, it was taken as positive; when it was <10%, it was regarded as negative. ② Estimation criteria of PCNA: It was observed for 5 successive high-fold microscopic fields for the percentage of the positive cells indicated by yellowish brown nucleus due to staining in each field, and the average value of the five fields was taken as labeling index (LI) for proliferated nuclear antigens.MAIN OUTCOME MEASURES: The expression levels of p21waf1, p53 and PCNA in lung cancer.RESULTS: ① The positive expression rates of p21waf1 and p53 in lung cancer were 75% (57/76) and 47%(36/76) respectively. The labeling index of PCNA in lung cancer group was significantly higher than that of the chronic pulmonary diseases group (0.44±0.32 vs 0.09±0.14, respectively).② Expression of p21waf1, p53 and PCNA was not related to tumor stages and tumor differentiation.③The LI of PCNA of squamous carcinoma was significantly higher than that of small cell lung cancer (0.51±0.33 vs 0.30±0.36). ④ Expression of p21waf1 or p53 had no relation with PCNA in lung cancer tissues. In chronic pulmonary diseases group, the expression of p21waf1 and p53 showed a close relationship with PCNA.CONCLUSION: It was found that p21waf1 and p53 were obviously upregulated in lung cancer and the degree of cellular proliferation in lung cancer was rather high. The capacity of DNA damage repair in squamous lung cancer may be stronger than that in small cell lung cancer. It was also found that p21waf1, p53 and PCNA collaborated with one another during the process of cellular proliferation in chronic pulmonary diseases, but not in lung cancer.更多还原