【作者】 弭丽丽；

【导师】 殷飞；

【作者基本信息】 河北医科大学 ， 内科学， 2010， 硕士

【摘要】 目的:Notch信号通路是一个在进化中高度保守的反映细胞间通讯机制的通路,它调控细胞的增殖、分化和凋亡,在胚胎发育和细胞命运的决定中发挥重要作用。Notch信号通路在不同组织细胞中作用不同,甚至在同一组织细胞的不同发展阶段作用也不同,这种生理作用依赖于细胞所处的环境。Notch信号不仅对正常细胞分化起重要作用,在肿瘤的形成中所起的重要作用已成为近年来研究的热点问题,但Notch信号在肝癌形成过程的机制,目前尚不十分清楚。本实验研究Notch信号配体分子Notch1-4和Jagged1在肝细胞癌、癌旁硬化肝组织及正常肝组织中的表达特点,并分析其与肝癌临床病理学参数的相关性,探讨这五种蛋白间的相关性。方法:1研究对象:50例肝细胞癌癌组织、相对应50例癌旁硬化肝组织及12例肝血管瘤周边正常肝组织均来自河北医科大学第四医院肝胆外科2008年5月~2009年4月手术切除标本。所有组织类型均经河北医科大学第四医院病理科确诊。所有患者术前均未进行任何形式的治疗。2研究方法:组织切除后经10%中性甲醛固定后石蜡包埋备用。应用免疫组织化学技术检测Notch1、Notch2、Notch3、Notch4及Jagged1在肝细胞癌癌组织、癌旁硬化肝组织及正常肝组织中的表达情况,分析染色强度在这三种肝组织的表达差异及其在临床病理学参数不同分组之间的差别。结果: 1免疫组化结果显示所检测的Notch1分子在各组肝组织均有表达,Notch1表现为细胞浆黄染。Notch1在肝癌组织的表达显著高于癌旁硬化肝组织和正常肝组织(Z=-4.004, P=0.000和Z=-3.355,P=0.001),癌旁硬化肝组织表达和正常肝组织表达无统计学差异。Notch1在肝癌组织的表达与患者性别、年龄、癌灶数目、肿瘤大小、AFP、HBsAg、HBeAg阴阳性、肿瘤包膜是否完整、瘤栓有无均无关;在肝癌病理分级高分化的癌组织中Notch1的表达强于低分化者(χ~2=6.938,P=0.031;r=-0.376,P=0.007);而Notch1的表达与肝癌临床分期无关。2 Notch2在各组肝组织均有表达,表现为细胞浆黄染。Notch2在肝癌组织、癌旁硬化肝组织、正常肝组织,这三者之间的表达无统计学差异。Notch2在肝癌组织的表达与患者性别、年龄、癌灶数目、肿瘤大小、AFP、HBsAg、HBeAg阴阳性、肿瘤包膜是否完整、瘤栓有无、肝癌临床分期及病理分级均无关。3 Notch3在各组肝组织均有表达,表现为细胞浆黄染。Notch3在肝癌组织的表达显著高于癌旁硬化肝组织和正常肝组织(Z=-2.551, P=0.011和Z=-2.436,P=0.015),在癌旁硬化肝组织表达和正常肝组织表达无统计学差异。Notch3在肝癌组织的表达与患者性别、年龄、癌灶数目、肿瘤大小、AFP、HBsAg、HBeAg阴阳性、肿瘤包膜是否完整、瘤栓有无均无关;在肝癌临床分期较晚的癌组织Notch3的表达强于分期较早者(χ~2=10.872,P=0.028;r=0.359,P=0.010);而Notch3的表达程度与肝癌病理分级无关。4 Notch4在各组肝组织均有表达,表现为细胞浆黄染。Notch4在肝癌组织的表达显著高于癌旁硬化肝组织和正常肝组织(Z=-2.906, P=0.004和Z=-2.985,P=0.003),癌旁硬化肝组织表达和正常肝组织表达无统计学差异。Notch4在肝癌组织的表达与患者性别、年龄、癌灶数目、肿瘤大小、AFP、HBsAg、HBeAg阴阳性、肿瘤包膜是否完整、瘤栓有无、肝癌临床分期及病理分级均无关。5 Jagged1在各组肝组织均有表达,表现为细胞浆黄染。Jagged1在肝癌组织的表达显著高于癌旁硬化肝组织和正常肝组织(Z=-2.747, P=0.006和Z=-2.249,P=0.025),癌旁硬化肝组织表达和正常肝组织表达无统计学差异。Jagged1在肝癌组织的表达与患者性别、年龄、癌灶数目、肿瘤大小、AFP、HBsAg、HBeAg阴阳性、肿瘤包膜是否完整、瘤栓有无、肝癌临床分期及病理分级均无关。6肝癌组织中Notch1、Notch2、Notch3、Notch4与Jagged1表达情况的相关性:肝癌组织中Notch1与Notch2的表达无相关性(r=-0.003,P=0.981),与Notch3的表达无相关性(r=0.166,P=0.249)与Notch4的表达无相关性(r=0.184,P=0.200),与Jagged1的表达无相关性(r=-0.248,P=0.082),Notch2与Notch3的表达无相关性(r=-0.054,P=0.709),与Notch4的表达无相关性(r=0.137,P=0.344)与Jagged1的表达无相关性(r=0.144,P=0.317),Notch3与Notch4的表达无相关性(r=0.138,P=0.339),与Jagged1的表达无相关性(r=0.166,P=0.249),Notch4与Jagged1的表达无相关性(r=0.245,P=0.086)结论:1 Notch1、Notch3、Notch4及Jagged1在肝细胞肝癌组织中的表达较癌旁硬化肝组织和正常肝组织升高,而在癌旁硬化肝组织和正常肝组织的表达无显著性差异。2 Notch1在肝癌病理分级高分化的癌组织中的表达较低分化者升高; Notch3在肝癌临床分期较晚的癌组织的表达较分期较早者升高,提示Notch信号通路的异常激活可能与肝癌的发生发展有关。更多还原

【Abstract】 Objective: Notch signaling pathway is an evolutionarily highly conserv- ed mechanism for cell-cell communication,which controls cell proliferation, differentiation,apoptosis,and plays an important role in embryogenesis and many types of cell fate determination.Notch signaling pathway has different function in different tissue cells and has different function at distinct stages of the same tissue cells.The biological function of this pathway is critically context dependent.Reports had showed that Notch signaling pathway is not only involved in the development of normal cells but also in cancer cells.Until now,the role of Notch signaling in HCC and its underlying mechanism are still unknown.To investigate the expression of Notch1-4 and Jagged1 in human hepatocellular carcinoma(HCC),adjacent non-cancerous cirrhotic hepatic tissue and normal hepatic tissue,we analyzed the association of their expression with clinicopathological parameters and the correlation among Notch1-4 and Jagged1.Methods: 1 Samples collection follow up: 50 specimens of HCC,50 specimens of adjacent non-cancerous cirrhotic hepatic tissue and 12 specimens of normal hepatic tissue were obtained from the fourth hospital of Hebei Medical University,which were not treated before operation.2 Imunostaining:The samples were fixed by 10% formaldehyde,embeded in paraffin after excision.Immunohistchemical staining was used to measure Notch1-4 and Jagged1 in HCC tissues and adjacent non-cancerous cirrhotic hepatic tissues and normal hepatic tissues,the differences of clinicopathologi- cal groups for Immunohistochemical staining were analyzed by statistical methods.Resluts: 1 Notch1 was expressed in HCC tissues,adjacent non-cancer- ous cirrhotic hepatic tissues,and normal hepatic tissues,which mostly expressed in cytoplasm.Notch1 expression in HCC tissue was much stronger than that in adjacent non-cancerous cirrhotic hepatic tissue and in normal hepatic tissue (Z=-4.004,P=0.000 and Z=-3.355,P=0.001),There was no statistical difference existed between in adjacent non-cancerous cirrhotic hepatic tissue and in normal hepatic tissue.Notch1 expression was significant stronger in better differentiation than that in poorer differentiation of HCC tissues (χ~2=6.938,P=0.031,r=-0.376,P=0.007).Notch1 expression in HCC tissues had showed no difference in different clinical stage.The Notch1 expression had no difference refered to sex,age,the number of tumor foci,size of tumor,AFP,HBsAg,HBeAg,the integrity of the tumor’s envelope and the portal vein tumor thrombosis in HCC tissues based on our statistical analysis.2 Notch2 was expressed in HCC tissues,adjacent non-cancerous cirrhotic hepatic tissues and normal hepatic tissues,which mostly expressed in cytoplasm.Notch2 expression had no statistical difference among HCC tissue,adjacent non-cancerous cirrhotic hepatic tissue and normal hepatic tissue.The Notch2 expression had no difference refered to sex,age,the number of tumor foci,size of tumor,AFP,HBsAg,HBeAg,the integrity of the tumor’s envelope,the portal vein tumor thrombosis,clinical stage,the pathological classification in HCC based on our statistical analysis.3 Notch3 was expressed in HCC tissues,adjacent non-cancerous cirrhotic hepatic tissues and normal hepatic tissues,which mostly expressed in cytoplasm.Notch3 expression in HCC tissue was much stronger than that in adjacent non-cancerous cirrhotic hepatic tissue and in normal hepatic tissue (Z=-2.551,P=0.011and Z=-2.436, P=0.015),There was no statistical difference existed between in adjacent non-cancerous cirrhotic hepatic tissue and in normal hepatic tissue.Notch3 expression was significant stronger in later clinical stage of HCC than earlier stage of HCC(χ~2=10.872,P=0.028,r=0.359, P=0.010), Notch3 expression showed no difference in different pathological classification of HCC. The Notch3 expression had no difference refered to sex,age,the number of tumor foci,size of tumor,AFP,HBsAg,HBeAg,the integrity of the tumor’s envelope and the portal vein tumor thrombosis in HCC based on our statistical analysis.4 Notch4 was expressed in HCC tissues,adjacent non-cancerous cirrhotic hepatic tissues and normal hepatic tissues,which mostly expressed in cytoplasm.Notch4 expression in HCC tissue was much stronger than that in adjacent non-cancerous cirrhotic hepatic tissue and in normal hepatic tissue (Z=-2.906,P=0.004 and Z=-2.985,P=0.003),There was no statistical difference existed between in adjacent non-cancerous cirrhotic hepatic tissue and in normal hepatic tissue.The Notch4 expression had no difference refered to sex,age,the number of tumor foci,size of tumor,AFP,HBsAg,HBeAg,the integrity of the tumor’s envelope,the portal vein tumor thrombosis,clinical stage,the pathological classification in HCC based on our statistical analysis.5 Jagged1 was expressed in HCC tissues,adjacent non-cancerous cirrhotic hepatic tissues and normal hepatic tissues,which mostly expressed in cytoplasm.Jagged1 expression in HCC tissue was much stronger than that in adjacent non-cancerous cirrhotic hepatic tissue and in normal hepatic tissue (Z=-2.747,P=0.006 and Z=-2.249,P=0.025),There was no statistical difference existed between in adjacent non-cancerous cirrhotic hepatic tissue and in normal hepatic tissue.The Jagged1 expression had no difference refered to sex,age,the number of tumor foci,size of tumor,AFP,HBsAg,HBeAg,the integrity of the tumor’s envelope,the portal vein tumor thrombosis,clinical stage,the pathological classification in HCC based on our statistical analysis.6 The correlation between Notch1,Notch2,Notch3,Notch4 and Jagged1 expression in HCC:Notch1 expression had no correlation with Notch2 expression level in HCC (r=-0.003,P=0.981),Notch1 expression level had no correlation with notch3 expression in HCC (r=0.166,P=0.249),Notch1 expression had no correlation with Notch4 expression level in HCC (r=0.184, P=0.200),Notch1 expression had no correlation with Jagged1 expression in HCC(r=-0.248,P=0.082),Noth2 expression had no correlation with Notch3 expression in HCC (r=-0.054, P=0.709),Notch2 expression had no correlation with Notch4 expression in HCC (r=0.137,P=0.344),Notch2 expression had no correlation with Jagged1 expression in HCC (r=0.144, P=0.317),Noth3 expressional level had no correlation with Notch4 expression in HCC (r=0.138,P=0.339),Noth3 expression had no correlation with Jagged1 expression in HCC (r=0.166,=0.249),Nothc4 expression had no correlation with Jagged1 expression in HCC (r=0.245,P=0.086).Conclusions: 1 Notch1,Notch3,Notch4 and Jagged1 expression level in HCC tissue was much higher than that in adjacent non-cancerous cirrhotic hepatic tissue and in normal hepatic tissue,but had no statistical difference existed between in adjacent non-cancerous cirrhotic hepatic tissue and in normal hepatic tissue.2 Notch1 expression level was significantly higher in better differentiation than poorer differentiation of HCC tissues,Notch3 expression level was significantly higher in later clinical stage of HCC than earlier stage,which showed that the abnormal activition of Notch signaling pathway might take effects on the progress of HCC.更多还原