【作者】 姚坦；

【导师】 孟翔凌； 杨文奇； 徐阿曼；

【作者基本信息】 安徽医科大学 ， 外科学， 2007， 硕士

【摘要】 背景与目的胆囊癌是胆道系统最常见的恶性肿瘤之一。胆囊癌的预后极差,手术治疗仍是目前治疗胆囊癌的主要方法。由于肿瘤的生物学异质性,仅依靠临床病理分期来评估预后有一定的局限性。细胞周期调控异常是细胞周期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,细胞周期调控异常,尤其是G1/S期调控点的异常,在肿瘤发生中起着很重要的作用。P27是近年来发现的一个重要的CDKI,主要通过抑制G1期的Cyclin-CDK的活性,使细胞停滞于G1期。CyclinE蛋白在细胞周期G1/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。P27作为一种抑癌基因,CyclinE作为一种癌基因,二者均作用于细胞周期G1/S期调控点而调节细胞的生长和分化。在肿瘤治疗的研究中癌基因、抑癌基因及其对细胞周期调控作用有着非常重要的意义。本课题主要通过研究P27和CyclinE在胆囊癌组织中的表达情况,探讨P27和CyclinE的表达与胆囊癌的发生、组织学分化程度、Nevin分期进展、肿瘤浸润深度、淋巴结转移和周围浸润等临床病理因素的关系,分析它们在胆囊癌发生、发展中的作用,进而揭示其在临床应用中的前景。方法收集安徽医科大学第一附属医院及六安市人民医院2001年1月～2006年12月收治的经手术切除且临床资料完整的胆囊癌标本40例。收集慢性胆囊炎标本20例做为对照组。标本经福尔马林固定,石蜡包埋,切片后用免疫组织化学法(SP法)检测P27和CyclinE的表达状况。对结果进行统计学分析,揭示P27和CyclinE的表达与胆囊癌及其各临床因素的相互关系。结果(1)慢性胆囊炎组织中P27的高表达阳性率为100%(20/20),明显高于胆囊癌的45%(18/40),(P<0.05)。CyclinE在胆囊癌组织中表达阳性率为67.5%(27/40),显著高于慢性胆囊炎的15.0%(3/20),(P<0.05)。(2)P27的表达与胆囊癌组织学分化程度、Nevin分期、肿瘤浸润深度、淋巴结转移、周围浸润有关(P<0.05),而与患者的性别、年龄、组织学类型、肿瘤大小无关(P>0.05)。CyclinE的表达与胆囊癌组织学分化程度、Nevin分期进展、肿瘤浸润深度有关(P<0.05),而与性别、年龄、组织学类型、肿瘤大小、淋巴结转移、周围浸润无关(P>0.05)。P27和CyclinE的表达与胆囊癌患者的预后明显相关(P<0.05)。P27可能成为胆囊癌的一个独立的预后指标物。(3)P27和CyclinE表达之间呈负相关关系(r=-0.375,P<0.05)。结论P27表达下降和CyclinE阳性表达导致细胞周期的调控异常,可能参与了胆囊癌的发生、发展过程。更多还原

【Abstract】 Background and objective Gallbladder carcinoma is one of the most popular malignant tumor in bile duct system. The prognosis of gallbladder carcinoma is very poor. Operation is the main treatment of gallbladder carcinoma. Histopathological stage is still the most popular indexes to predict patient’ s prognosis. However, tumor stage may lead physician to a wrong way sometimes because of the biological heterogeneity of tumor. Recent studies have revealed that uncontrolled cell cycle is very important to uncontrolled cell proliferation and carcinogenesis. Cyclin-dependent kinase(CDK),together with cylin and CDK inhibitor, govern cell cycle progression in eukaryotic cell. In cell cycle, different phase is regulated by different factors. Abnormal control on cell cycle, especially the abnorm of the check point of G1/S stage, plays an important role in carcinogenesis.. P27 is an important CDK inhibitor, which binds to and inhibits preferentially G1-phase kinases thereby arresting the cell cycle at G1-phase. Proto-oncogene cyclinE, encodes cyclinE protein,which acts as an important positive regulator in G1/S transition,and appears to be closely linked with carcinogenesis. CyclinE,as an oncogene and p27, as an antioncogene modulate the growth and diferentiation of cells by working on the cell cycle check point of G1/S stage, role in carcinogenesis. The researches on oncogene, antioncogene and the role of cell cycle control have great significance to help us treat tumor. This research, by studying the expressions of cyclinE and p27 in the tissues of gallbladder carcinomas (GBC), was to explore the correlation of cyclinE and p27 and gallbladder carcinogenesis,the histological grade, nevin′s stage, depth of invasion, and hence evaluate their roles in carcinogenesis and their significance at clinic.Methods The samples from 40 patients with gallbladder carcinoma, who underwent surgical resection at The First Afiliated Hosptial of Anhui Medical University and Lu’An Peaple’s Hospital from 2001 Jan to 2006 Dec, were used for this study. 20 chronic cholecystitis (CC) specimens were control group. The samples were routinely fixed in 10% formalin and embedded in paraffin. After sectioned into slices, we detected the expression of p27 and cyclinE protein by immunohistochemical staining technique(SP method). We analyzed the relationship between the expression of p27 and cyclinE protein with clinical parameters.Results (1)The positive expression rate of p27 in gallbladder carcinoma was 45.0%(18/40), which was lower than that in chronic cholecystitis(100% ), P<0.05. The positive expression rate of cyclinE in gallbladder carcinoma was 67.5%(27/40), which was significantly higher than that in chronic cholecystitis(15.0%,3/20), P<0.05. (2)The expression level of p27 protein in gallbladder carcinomas showed an obvious difference in the histological grade, nevin′s stage, depth of invasion, lymph node metastasis, infiltration (P<0.05). But not markedly associated with patient′s age, , histological types, tumor size. The expression level of cyclinE protein in gallbladder carcinomas showed an obvious difference in the histological grade,nevin′s stage, depth of invasion (P<0.05). But not markedly associated with patient′s age, histological types, tumor size, lymph node metastasis, infiltration. Log rank test showed that the p27 and cyclinE expression were significantly related with prognosis of the patients (P<0.05).(3)There was negative correlation between p27 and cyclinE expression (r=-0.375,P<0.05).Conclusion The decreased expression of p27 and the high expression of cyclin E result in abnomal regulation of cell cycle,which may be associated with gallbladder carcinogenesis and progression.更多还原