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小鼠心肌組織裂解上清液誘導(dǎo)小鼠骨髓瘤Sp2/0細(xì)胞凋亡
作者：王瑋，張文艷，吳建軍，類延花，金晶，方草暉，胡勇，王明麗【關(guān)鍵詞】 心肌上清液
Apoptosis of mouse myeloma Sp2/0 cells induced by supernatant of mouse cardiac muscle lysate
【Abstract】 AIM: To study the roles of mouse cardiac muscle lysate supernatant in inducing the apoptosis of mouse myeloma Sp2/0 cells. METHODS: Various concentrations (1∶6, 1∶12, 1∶24) of mouse cardiac muscle lysate supernatant and cyclophosphamide were added into the Sp2/0 cell culture wells. After 24 h, microscope was used to observe the morphological changes of Sp2/0 cells and apoptosis rate was determined by flow cytometry after 48 h. Morphological changes of Sp2/0 cells cocultured with mouse cardiac muscle cells were observed after 10 h. The apoptosis of Sp2/0, Hela, Hep2 and Vero cells was induced with various concentrations (1∶5, 1∶10, 1∶20 and 1∶40) of mouse cardiac muscle lysate supernatant, liver lysate supernatant, thymus lysate supernatant and cyclophosphamide, and then the difference of apoptotic rate among them was compared. RESULTS: Mouse cardiac muscle lysate supernatant could induce the apoptosis of the Sp2/0 cells and the apoptotic rate increased in a concentrationdependent manner, but there was no difference between mouse cardiac muscle lysate supernatant group and cyclophosphamide group. After a 10hour coculture with mouse cardiac muscle cell lysate, the apoptosis of a few Sp2/0 cells was seen. The mouse cardiac muscle lysate supernatant, liver lysate supernatant and thymus lysate supernatant could all induce apoptosis of the Sp2/0, HeLa, Hep2 and Vero cells. But the effect of the mouse cardiac muscle lysate supernatant was more obvious and permanent. CONCLUSION: The mouse cardiac muscle lysate supernatant can significantly induce apoptosis of mouse myeloma Sp2/0 cells.
【Keywords】 cardiac muscle lysate supernatant; apoptosis; mouse myeloma Sp2/0 cell
【摘要】 目的: 研究小鼠心肌上清液誘導(dǎo)小鼠骨髓瘤Sp2/0細(xì)胞凋亡的作用. 方法： 在Sp2/0細(xì)胞培養(yǎng)孔中，分別加入不同稀釋度(1∶6, 1∶12, 1∶24)的小鼠心肌上清液和環(huán)磷酰胺， 24 h后鏡下觀察細(xì)胞形態(tài)，并于48 h后流式細(xì)胞儀檢測(cè)凋亡細(xì)胞的比率；同時(shí)，Sp2/0細(xì)胞和小鼠心肌細(xì)胞共培養(yǎng)，10 h后觀察Sp2/0細(xì)胞形態(tài)學(xué)變化；并用不同稀釋度（1∶5, 1∶10, 1∶20, 1∶40）小鼠心肌裂解上清液、肝臟裂解上清液、胸腺裂解上清液及環(huán)磷酰胺誘導(dǎo)Sp2/0, HeLa, Hep2和Vero細(xì)胞凋亡，并比較其差異. 結(jié)果: 不同稀釋度的心肌裂解上清液均可誘導(dǎo)Sp2/0細(xì)胞的凋亡，尤以高稀釋度更明顯；小鼠心肌細(xì)胞與Sp2/0細(xì)胞共培養(yǎng)10 h后,少部分Sp2/0細(xì)胞出現(xiàn)凋亡；小鼠心肌裂解上清液、肝臟裂解上清液、胸腺裂解上清液均可引起Sp2/0, HeLa, Hep2和Vero細(xì)胞發(fā)生凋亡，但小鼠心肌裂解上清液作用明顯，且作用時(shí)間持久. 結(jié)論: 小鼠心肌組織裂解上清液具有明顯誘導(dǎo)小鼠骨髓瘤Sp2/0細(xì)胞凋亡的作用.
【關(guān)鍵詞】 心肌上清液；細(xì)胞凋亡；小鼠骨髓瘤Sp2/0細(xì)胞
0引言
細(xì)胞凋亡與腫瘤的發(fā)生發(fā)展密切相關(guān)，已成為研究熱點(diǎn)［1］，許多預(yù)防治療腫瘤的藥物、細(xì)胞因子等抑制腫瘤細(xì)胞生長(zhǎng)的機(jī)制之一是誘導(dǎo)腫瘤細(xì)胞凋亡. 我們從細(xì)胞凋亡的角度探討心肌上清液誘導(dǎo)小鼠骨髓瘤Sp2/0細(xì)胞凋亡.
1材料和方法
1.1材料
Balb/c 20只8周齡小鼠，雌雄各半，體質(zhì)量約250 g，由安徽安科生物工程股份有限公司動(dòng)物室提供；環(huán)磷酰胺(2 g/L)由湖北科利藥業(yè)股份有限公司生產(chǎn); DMEM液由美國(guó)生命技術(shù)公司生產(chǎn)；胎牛血清由杭州四季青公司提供；流式細(xì)胞儀（日本產(chǎn)）；Leica熒光顯微鏡(德國(guó)Leica公司)；CO2溫箱(美國(guó)產(chǎn))；紫外分光光度計(jì)（上海產(chǎn)）. Sp2/0, HeLa, Hep2和Vero細(xì)胞株均來源于中國(guó)預(yù)防科學(xué)院病毒所，均以100 mL/L胎牛血清＋DMEM液為培養(yǎng)液，按本室常規(guī)方法在CO2溫箱中培養(yǎng)；乳鼠心肌細(xì)胞培養(yǎng)： 取Balb/c新生紅皮乳鼠20只，在無(wú)菌操作下取出心臟，用PBS沖洗3遍，以眼科剪剪成1 mm3碎塊，洗滌，后加2.5 g/L胰蛋白酶液37℃消化20 min，吹打3次，吸取上層細(xì)胞懸液，以同樣的方法消化3次，直到組織塊完全消化為止，以1500 r/m
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