【摘要】：目的:探討交聯(lián)CD44分子對人肥大細胞白血病HMC-1細胞株Fas表達及其介導的細胞凋亡的影響。方法:應(yīng)用流式細胞術(shù)檢測HMC-1細胞表面CD44及Fas的表達,并觀察透明質(zhì)酸(native hyaluronan,HA)、低分子質(zhì)量透明質(zhì)酸(fragmented hyaluronan,F-HA)處理或抗體交聯(lián)CD44分子后細胞表面Fas表達的變化,進而檢測PI3K、PKC及MAPK信號通路抑制劑、蛋白質(zhì)合成抑制劑和肌動蛋白聚合抑制劑對CD44分子交聯(lián)后細胞表面Fas表達的影響,用Northern雜交技術(shù)檢測交聯(lián)CD44對Fas mRNA表達的影響,用Annexin V/PI雙染法檢測交聯(lián)CD44、HA或F-HA對Fas介導的細胞凋亡的影響。結(jié)果:HMC-1細胞高表達CD44而低表達Fas,交聯(lián)CD44分子顯著上調(diào)細胞表面Fas的表達(P0.05),而Fas mRNA轉(zhuǎn)錄水平?jīng)]有明顯變化;肌動蛋白聚合抑制劑細胞松弛素B能抑制CD44上調(diào)的Fas表達(P0.05),所檢測細胞信號通路抑制劑和蛋白質(zhì)合成抑制劑均未能抑制Fas表達的上調(diào)(P0.05);F-HA和交聯(lián)CD44均能夠促進Fas介導的細胞凋亡(P0.05)。結(jié)論:CD44分子可上調(diào)HMC-1細胞的Fas表達并放大Fas介導的細胞凋亡,CD44分子可能成為肥大細胞白血病治療的新靶點。
[Abstract]:Aim: to investigate the effect of cross-linked CD44 on Fas expression and apoptosis in HMC-1 cells of human mast cell leukemia. Methods: flow cytometry was used to detect the expression of CD44 and Fas on the surface of HMC-1 cells, and to observe the expression of native hyaluronan,HA and (fragmented hyaluronan, of low molecular weight hyaluronic acid. The changes of Fas expression on cell surface were detected after F-HA treatment or antibody crosslinking CD44 molecule, and then PI3K,PKC and MAPK signal pathway inhibitors were detected. The effects of protein synthesis inhibitor and actin polymerization inhibitor on the expression of Fas on the cell surface after CD44 molecular crosslinking were studied. The effect of cross-linked CD44 on Fas mRNA expression was detected by Northern hybridization, and cross-linked CD44, was detected by Annexin V/PI double staining. Effect of HA or F-HA on apoptosis mediated by Fas. Results: high expression of CD44 and low expression of Fas, crosslinked CD44 in HMC-1 cells significantly up-regulated the expression of Fas on the cell surface (P0.05), while the level of Fas mRNA transcription did not change significantly. Actin polymerization inhibitor cytochalasin B could inhibit the up-regulated Fas expression of CD44 (P0.05). Neither the signal pathway inhibitor nor protein synthesis inhibitor could inhibit the up-regulation of Fas expression (P0.05). Both F-HA and CD44 could promote apoptosis mediated by Fas (P0.05). Conclusion: CD44 can up-regulate the expression of Fas in HMC-1 cells and amplify the apoptosis mediated by Fas. CD44 may be a new target for the treatment of mast cell leukemia.
【作者單位】： 河北醫(yī)科大學第四醫(yī)院腫瘤研究所免疫室;河北醫(yī)科大學第四醫(yī)院科研中心;產(chǎn)業(yè)醫(yī)科大學附屬病院第一內(nèi)科;
【基金】：國家自然科學基金資助項目(No.81402228) 河北省自然科學基金資助項目(No.H2015206216) 河北省醫(yī)學基金資助項目(No.ZL20140334) 河北省教育基金資助項目(No.QN2014049)~~
【分類號】：R733.7

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