目的:探讨阿霉素对口腔鳞癌干细胞迁移、侵袭、凋亡的影响及其可能的机制。方法:体外培养人口腔鳞癌细胞系SCC25,通过流式细胞术分选CD44-和CD44+细胞,RT-PCR检测CD44-和CD44+细胞的Oct4、CD133、CD44和GAPDH的m RNA表达;检测和比较CD44-和CD44+细胞的克隆形成能力。CD44+细胞用阿霉素或β-catenin抑制剂LF3进行处理,分别使用Transwell和细胞划痕检测细胞侵袭和迁移能力,一步法TUNEL检测细胞凋亡水平,WB检测β-catenin和TCF-4的蛋白表达。结果:流式细胞术成功分离CD44-和CD44+细胞,RT-PCR检测CD44+细胞高表达Oct4、CD133和CD44 m RNA,CD44-细胞弱表达Oct4m RNA,不表达CD133和CD44 m RNA;CD44+细胞的克隆形成能力显示显著强于CD44-细胞(P<0.05)。阿霉素显著降低了CD44+细胞的侵袭能力和迁移能力(P<0.05),显著提高了CD44+细胞的凋亡率(P<0.05);阿霉素显著降低了CD44+细胞β-catenin和TCF-4的蛋白表达(P<0.05),LF3对β-catenin和TCF-4蛋白表达的影响与阿霉素比较无显著差异(P>0.05)。结论:阿霉素可能通过抑制Wnt/β-catenin信号通路降低口腔鳞癌干细胞迁移、侵袭能力,促进细胞凋亡。 相似文献
The park management (PM) that evolved from the park-renewal project based on Installation-Management Permission (IMP) in Tennoji Park, a major urban park in Osaka City, Japan is evaluated herein. The PM of Tennoji Park is composed of 'hard' and 'soft' tasks. The process and characteristics of the hard tasks, i.e., the park-renewal project including the construction of a significant landmark, the lawn plaza (named 'TEN-SHIBA') and convenience facilities with various service functions are analyzed. The details and outcomes of the soft tasks including cleaning, security, and lawn/planting management plus events held on the TEN-SHIBA plaza are also clarified. The PM results and financial effects of the introduction of IMP in TEN-SHIBA are evaluated based on both the park users' characteristics obtained by a questionnaire, an observation survey of the park users, and a data analysis of the track record. Based on a comparison with Sumiyoshi Park, an urban park similar to TEN-SHIBA but without the introduction of IMP, it was found that young females were the predominant users of TEN-SHIBA, and the average staying time on the lawn plaza was 2 min. longer than that in Sumiyoshi park. Together these results suggest the effectiveness of IMP for PM based on the TEN-SHIBA experience. It also appears that placing a plaza in the center of a park and providing facilities and various events by private business operators can generate profits; this might be effective for future PM hard tasks, while various and continuous soft task efforts remain essential.
Several models of activation mechanisms were proposed for G protein-coupled receptors (GPCRs), yet no direct methods exist for their elucidation. The availability of constitutively active mutants has given an opportunity to study active receptor conformations within acceptable limits using models such as the angiotensin II type 1 (AT1)1 receptor mutant N111G-hAT1 which displays an important constitutive activity. Recently, by using methionine proximity assay, we showed for the hAT1 receptor that TMD III, VI, and VII form the ligand-binding pocket of the C-terminal amino acid of an antagonistic AngII analogue. In the present contribution, we investigated whether the same residues would also constitute the ligand-binding contacts in constitutively activated mutant (CAM) receptors. For this purpose, the same Met mutagenesis strategy was carried out on the N111G double mutants. Analysis of 43 receptors mutants in the N111G-hAT1 series, photolabeled and CNBr digested, showed that there were only subtle structural changes between the wt-receptor and its constitutively active form. 相似文献