为了在建立的PC-12细胞肿瘤移植模型基础上,对不同时间点肿瘤组织中的Caspase-3和Caspase-9的表达进行检测,探讨Caspase-3和Caspase-9在肿瘤发展过程中的作用和意义,试验将指数生长期的PC-12细胞以0.1 m L(1×107个/m L)的密度接种...为了在建立的PC-12细胞肿瘤移植模型基础上,对不同时间点肿瘤组织中的Caspase-3和Caspase-9的表达进行检测,探讨Caspase-3和Caspase-9在肿瘤发展过程中的作用和意义,试验将指数生长期的PC-12细胞以0.1 m L(1×107个/m L)的密度接种于出生第7天的昆明白小鼠右前肢腋窝皮下,在接种后的不同时间点(7,14,21天)分别进行取材,将肿瘤块及邻近组织一并取下,通过多聚甲醛固定、包埋剂包埋后进行冰冻切片,采用荧光免疫组织化学法对Caspase-3和Caspase-9的表达进行检测。结果表明:刚出生7天的昆明白小鼠,接种PC-12细胞后7天时便可以明显观察到肿瘤块的形成,在14天时肿瘤块最大,21天时肿块减小,到28天时肿瘤块基本消失;对照组小鼠肌肉组织中Caspase-3和Caspase-9都呈现低表达。在移植肿瘤7天的肿块实体组织中,Caspase-3和Caspase-9都呈阳性表达;在14天时实体组织中表达降低,但是在周围组织中的表达增强;在21天时周围组织的表达也减弱。说明普通昆明白小鼠幼鼠中不能够建立真正的肿瘤模型,但是短期内仍能形成瘤块,在不同时间点,Caspase-3和Caspase-9表达具有明显的区域和量的差异,但是Caspase-3和Caspase-9表达部位和表达量的变化具有相似性。展开更多
AIM: To investigate the role of matrix-degrading metalloproteinases 9, 12(MMPs), as mediators of functional connective tissue damage in actinic cheilitis.METHODS: Thirty five formalin-fixed, paraffin embedded specimen...AIM: To investigate the role of matrix-degrading metalloproteinases 9, 12(MMPs), as mediators of functional connective tissue damage in actinic cheilitis.METHODS: Thirty five formalin-fixed, paraffin embedded specimens of actinic cheilitis, and twelve specimens of normal lower lip vermillion, which were obtained by the archives of the Department of Oral Medicine and Maxillofacial Pathology, were examined. From each block, 5 μm thick sections were cut and routinely stained with Hematoxylin and Eosin. Immunohistochemical studies were performed on 4-μm thick sections of formalin-fixed paraffin embedded actinic cheilitis lesions and of normal lower lip vermillion, for MMP-9 and MMP-12 in serial sections of our specimens. Appropriate positive and negative controls were performed to confirm the specificity of the staining reaction. MMP immunohistochemistry was evaluated using a semiquantitative immunoreactive score.RESULTS: Haematoxylin and eosin staining revealedin actinic cheilitis lesions atrophic stratified squamous cell epithelium, or focally and irregularly hyperplastic of variable thickness, in some areas was observed marked keratin production. Varying degrees of epithelial dysplasia were noticed with a wide spectrum of change within the same specimen. Characteristic was the appearance of chronic inflammatory infiltration, and a band of amorphous acellular, basophilic change like solar elastosis(elastin replacement of collagen). In normal lower lip specimens weak and scanty positive expression of MMP-9 and MMP-12 was observed. Anti-MMP-9 antibody showed a weak reaction, in actinic cheilitis lesions, focal in the elastotic material, in chronic inflammatory cells and mostly in macrophages and neutrophils. Strong and in some cases diffused immunohistochemical expression of MMP-12 was detected in actinic cheilitis lesions in the areas of the fragmented, distorted and thickened elastic fibers. MMP-12 was also expressed in chronic inflammatory cells and mostly macrophages. MMP-12 was significantly higher in actinic cheilitis specimens compared with the normal lower lip specimens(P = 0.0029).CONCLUSION: Our results suggest that especially MMP-12 may play an important role in remodeling events occurring in the connective tissue during long-term exposure to sunlight in the actinic cheilitis lesions.展开更多
文摘为了在建立的PC-12细胞肿瘤移植模型基础上,对不同时间点肿瘤组织中的Caspase-3和Caspase-9的表达进行检测,探讨Caspase-3和Caspase-9在肿瘤发展过程中的作用和意义,试验将指数生长期的PC-12细胞以0.1 m L(1×107个/m L)的密度接种于出生第7天的昆明白小鼠右前肢腋窝皮下,在接种后的不同时间点(7,14,21天)分别进行取材,将肿瘤块及邻近组织一并取下,通过多聚甲醛固定、包埋剂包埋后进行冰冻切片,采用荧光免疫组织化学法对Caspase-3和Caspase-9的表达进行检测。结果表明:刚出生7天的昆明白小鼠,接种PC-12细胞后7天时便可以明显观察到肿瘤块的形成,在14天时肿瘤块最大,21天时肿块减小,到28天时肿瘤块基本消失;对照组小鼠肌肉组织中Caspase-3和Caspase-9都呈现低表达。在移植肿瘤7天的肿块实体组织中,Caspase-3和Caspase-9都呈阳性表达;在14天时实体组织中表达降低,但是在周围组织中的表达增强;在21天时周围组织的表达也减弱。说明普通昆明白小鼠幼鼠中不能够建立真正的肿瘤模型,但是短期内仍能形成瘤块,在不同时间点,Caspase-3和Caspase-9表达具有明显的区域和量的差异,但是Caspase-3和Caspase-9表达部位和表达量的变化具有相似性。
文摘AIM: To investigate the role of matrix-degrading metalloproteinases 9, 12(MMPs), as mediators of functional connective tissue damage in actinic cheilitis.METHODS: Thirty five formalin-fixed, paraffin embedded specimens of actinic cheilitis, and twelve specimens of normal lower lip vermillion, which were obtained by the archives of the Department of Oral Medicine and Maxillofacial Pathology, were examined. From each block, 5 μm thick sections were cut and routinely stained with Hematoxylin and Eosin. Immunohistochemical studies were performed on 4-μm thick sections of formalin-fixed paraffin embedded actinic cheilitis lesions and of normal lower lip vermillion, for MMP-9 and MMP-12 in serial sections of our specimens. Appropriate positive and negative controls were performed to confirm the specificity of the staining reaction. MMP immunohistochemistry was evaluated using a semiquantitative immunoreactive score.RESULTS: Haematoxylin and eosin staining revealedin actinic cheilitis lesions atrophic stratified squamous cell epithelium, or focally and irregularly hyperplastic of variable thickness, in some areas was observed marked keratin production. Varying degrees of epithelial dysplasia were noticed with a wide spectrum of change within the same specimen. Characteristic was the appearance of chronic inflammatory infiltration, and a band of amorphous acellular, basophilic change like solar elastosis(elastin replacement of collagen). In normal lower lip specimens weak and scanty positive expression of MMP-9 and MMP-12 was observed. Anti-MMP-9 antibody showed a weak reaction, in actinic cheilitis lesions, focal in the elastotic material, in chronic inflammatory cells and mostly in macrophages and neutrophils. Strong and in some cases diffused immunohistochemical expression of MMP-12 was detected in actinic cheilitis lesions in the areas of the fragmented, distorted and thickened elastic fibers. MMP-12 was also expressed in chronic inflammatory cells and mostly macrophages. MMP-12 was significantly higher in actinic cheilitis specimens compared with the normal lower lip specimens(P = 0.0029).CONCLUSION: Our results suggest that especially MMP-12 may play an important role in remodeling events occurring in the connective tissue during long-term exposure to sunlight in the actinic cheilitis lesions.
