【摘要】：目的探討阿司匹林誘導(dǎo)EB病毒轉(zhuǎn)化的人B淋巴細(xì)胞凋亡及其機(jī)制。方法 EB病毒轉(zhuǎn)化的人B淋巴細(xì)胞經(jīng)一定濃度的阿司匹林處理后,首先通過MTT法分析細(xì)胞的增殖情況;然后用光學(xué)顯微鏡和電子顯微鏡、PI染色和流式細(xì)胞術(shù)分析以及瓊脂糖凝膠電泳等方法對(duì)細(xì)胞凋亡進(jìn)行評(píng)價(jià);最后,通過免疫印記法檢測凋亡相關(guān)蛋白、mTOR信號(hào)通路和PU.1-Bim信號(hào)通路蛋白表達(dá)情況。結(jié)果阿司匹林能抑制EB病毒轉(zhuǎn)化的人B淋巴細(xì)胞增殖。形態(tài)學(xué)和超微結(jié)構(gòu)觀察發(fā)現(xiàn),阿司匹林處理的細(xì)胞固縮變小、數(shù)目減少、細(xì)胞核畸形、染色質(zhì)邊緣化和細(xì)胞質(zhì)空泡化。阿司匹林處理導(dǎo)致細(xì)胞膜通透性增加、細(xì)胞存活率明顯下降和細(xì)胞DNA的彌散現(xiàn)象。免疫印跡分析顯示,阿司匹林處理抑制了細(xì)胞的mTOR信號(hào)和轉(zhuǎn)錄因子PU.1的表達(dá)水平,激活了細(xì)胞凋亡相關(guān)蛋白caspase-3、PARP和Bim。結(jié)論阿司匹林可能通過抑制EB病毒轉(zhuǎn)化的人B淋巴細(xì)胞增殖和誘導(dǎo)細(xì)胞凋亡表現(xiàn)一定的抗淋巴瘤效應(yīng),mTOR信號(hào)途徑和PU.1-Bim軸可能參與了阿司匹林抗腫瘤作用機(jī)制。
[Abstract]:Objective to investigate the apoptosis of human B lymphocytes transformed by Epstein-Barr virus (EBV) induced by aspirin and its mechanism. Methods Epstein-Barr virus transformed human B lymphocytes were treated with aspirin at a certain concentration. The proliferation of B lymphocytes was analyzed by MTT assay. Then the apoptosis was evaluated by optical microscope, electron microscope, Pi staining, flow cytometry and agarose gel electrophoresis. The protein expression of apoptosis-related protein (mTOR) and PU.1-Bim signal pathway (PU.1-Bim) were detected by immunoblotting. Results Aspirin could inhibit the proliferation of human B lymphocytes transformed by EB virus. Morphologic and ultrastructural observation showed that the cells treated with aspirin had smaller pyknosis, decreased number, nuclear malformation, chromatin marginalization and cytoplasmic vacuolation. Aspirin treatment resulted in increased cell membrane permeability, decreased cell survival and dispersion of cell DNA. Western blot analysis showed that aspirin treatment inhibited the expression of mTOR signal and transcription factor PU.1, and activated the apoptosis-related proteins caspase-3 PARP and Bim. Conclusion Aspirin may be involved in the anti-tumor mechanism of aspirin by inhibiting the proliferation of human B lymphocytes transformed by EB virus and inducing apoptosis.
【作者單位】： 中國醫(yī)學(xué)科學(xué)院北京協(xié)和醫(yī)學(xué)院醫(yī)學(xué)生物學(xué)研究所;
【基金】：國家自然科學(xué)基金資助項(xiàng)目(No81571549) 云南省應(yīng)用基礎(chǔ)研究計(jì)劃項(xiàng)目(No2013FZ143) 云南省重點(diǎn)新產(chǎn)品開發(fā)專項(xiàng)項(xiàng)目(No2016BC004) 醫(yī)學(xué)生物學(xué)研究所重點(diǎn)項(xiàng)目(No2014IMB03ZD)
【分類號(hào)】：R96

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