论文部分内容阅读
目的:探讨交联CD44分子对人肥大细胞白血病HMC-1细胞株Fas表达及其介导的细胞凋亡的影响。方法:应用流式细胞术检测HMC-1细胞表面CD44及Fas的表达,并观察透明质酸(native hyaluronan,HA)、低分子质量透明质酸(fragmented hyaluronan,F-HA)处理或抗体交联CD44分子后细胞表面Fas表达的变化,进而检测PI3K、PKC及MAPK信号通路抑制剂、蛋白质合成抑制剂和肌动蛋白聚合抑制剂对CD44分子交联后细胞表面Fas表达的影响,用Northern杂交技术检测交联CD44对Fas mRNA表达的影响,用Annexin V/PI双染法检测交联CD44、HA或F-HA对Fas介导的细胞凋亡的影响。结果:HMC-1细胞高表达CD44而低表达Fas,交联CD44分子显著上调细胞表面Fas的表达(P<0.05),而Fas mRNA转录水平没有明显变化;肌动蛋白聚合抑制剂细胞松弛素B能抑制CD44上调的Fas表达(P<0.05),所检测细胞信号通路抑制剂和蛋白质合成抑制剂均未能抑制Fas表达的上调(P>0.05);F-HA和交联CD44均能够促进Fas介导的细胞凋亡(P<0.05)。结论:CD44分子可上调HMC-1细胞的Fas表达并放大Fas介导的细胞凋亡,CD44分子可能成为肥大细胞白血病治疗的新靶点。
Objective: To investigate the effect of cross-linked CD44 on Fas expression and apoptosis-mediated apoptosis in human mast cell leukemia HMC-1 cell line. Methods: The expression of CD44 and Fas on the surface of HMC-1 cells was detected by flow cytometry. The effects of hyaluronan (HA), low-molecular-weight hyaluronan (F-HA) CD44 molecules on the cell surface Fas expression changes, and then detect PI3K, PKC and MAPK signal pathway inhibitors, protein synthesis inhibitors and actin polymerization inhibitors on CD44 molecules after cell surface Fas expression, using Northern hybridization The effect of cross-linked CD44 on Fas mRNA expression was examined. The effect of cross-linked CD44, HA or F-HA on Fas-mediated apoptosis was detected by Annexin V / PI double staining. RESULTS: HMC-1 cells highly expressed CD44 but had low expression of Fas. Cross-linking CD44 molecules significantly up-regulated the expression of Fas on the cell surface (P <0.05), while Fas mRNA transcription did not change significantly. Actin polymerization inhibitor cytochalasin B (P <0.05). Both the inhibitor of cell signaling pathway and inhibitor of protein synthesis did not inhibit the up-regulation of Fas expression (P> 0.05). Both F-HA and cross-linked CD44 could promote the expression of Fas Mediated apoptosis (P <0.05). CONCLUSION: CD44 can up-regulate the expression of Fas in HMC-1 cells and increase Fas-mediated apoptosis. CD44 may be a new target for the treatment of leukemia in mast cells.