發(fā)布時間：2018-05-29 13:19

  本文選題：來那度胺 + 阿司匹林　； 參考：《南昌大學》2015年碩士論文

【摘要】：目的:1、觀察來那度胺(lenalidomide,LEN)和阿司匹林(aspirin,ASA)聯(lián)合對骨髓瘤細胞株增殖、凋亡與細胞周期的影響,分析兩者在骨髓瘤細胞株中的相互作用。2、檢測LEN和ASA聯(lián)合對骨髓瘤細胞VEGF蛋白及wnt/β-catenin信號通路蛋白表達的影響,探討ASA對LEN抗骨髓瘤的增敏機制。方法:1、常規(guī)培養(yǎng)骨髓瘤細胞株(MM1.S及RPMI-8226),制作細胞生長曲線,選取對數(shù)生長期細胞進行后續(xù)實驗。2、采用CCK-8法檢測LEN和ASA聯(lián)合用藥對MM1.S細胞和RPMI-8226細胞增殖的影響。3、采用Annexin V-FITC/PI染色方法檢測LEN和ASA聯(lián)合用藥對MM1.S細胞和RPMI-8226細胞凋亡的影響。4、采用流式細胞儀檢測LEN和ASA聯(lián)合用藥對MM1.S細胞和RPMI-8226細胞周期的影響。5、Western Blot法檢測LEN和ASA聯(lián)合用藥對MM1.S細胞和RPMI-8226細胞VEGF蛋白、β-catenin及下游分子cyclinD1蛋白表達的影響。結果:1、藥物作用24~72 h后,LEN與ASA聯(lián)合以時間依賴性抑制細胞增殖活性,且顯著高于LEN(1μM)及ASA(2.5 mM)單藥組。相互作用分析顯示:LEN與ASA在抗MM細胞增殖中呈協(xié)同作用。2、藥物作用48 h后,LEN與ASA聯(lián)合組對MM1.S及RPMI-8226細胞凋亡誘導率顯著高于LEN或ASA單藥組。3、藥物作用48 h后,與LEN或ASA單藥組相比,LEN與ASA聯(lián)合組誘導MM1.S及RPMI-8226細胞阻滯于G1期比率明顯增高。4、藥物作用48 h后,相比LEN或ASA單藥組,兩者聯(lián)合后顯著下調了MM1.S及RPMI-8226細胞VEGF蛋白表達。5、在MM1.S與RPMI-8226細胞中,LEN作用72 h后誘導胞核內(nèi)β-catenin蛋白及其下游cyclin D1蛋白表達上調,而ASA促使胞漿β-catenin磷酸化,導致胞核內(nèi)β-catenin蛋白其下游分子cyclin D1蛋白表達下降。結論:1、阿司匹林與來那度胺在抑制骨髓瘤細胞增殖中呈協(xié)同作用;2、阿司匹林通過下調骨髓瘤細胞株VEGF表達對來那度胺發(fā)揮協(xié)同抗骨髓瘤作用;3、阿司匹林通過促進細胞漿β-catenin磷酸化,從而阻止β-catenin入細胞核發(fā)揮對來那度胺的化療增敏作用。
[Abstract]:Objective: to observe the effects of the combination of renalidomide en and aspirin on the proliferation, apoptosis and cell cycle of myeloma cell line. To investigate the effect of LEN and ASA on the expression of VEGF protein and wnt/ 尾 -catenin signal pathway protein in myeloma cell line, and to explore the mechanism of ASA on anti-myeloma. Methods: cell growth curves were prepared by routine culture of myeloma cell lines MM1.S and RPMI-8226. Logarithmic growth phase cells were selected for follow-up experiment. CCK-8 assay was used to detect the effect of combined use of LEN and ASA on the proliferation of MM1.S cells and RPMI-8226 cells. Annexin V-FITC/PI staining was used to detect the apoptosis of MM1.S cells and RPMI-8226 cells treated with LEN and ASA. The effect of combined use of LEN and ASA on the cell cycle of MM1.S and RPMI-8226 was detected by flow cytometry. The effects of LEN and ASA on the expression of VEGF protein, 尾 -catenin and cyclinD1 in MM1.S and RPMI-8226 cells were detected by Western Blot assay. Results the cell proliferation activity was inhibited in a time-dependent manner by the combination of Len and ASA for 24 ~ 72 h, and was significantly higher than that in LEN(1 渭 M and ASA(2.5 mm) groups. The results of interaction analysis showed that ASA and WLEN had synergistic effects on the proliferation of MM cells. After 48 h of drug treatment, the apoptotic induction rate of MM1.S and RPMI-8226 cells in the combination group was significantly higher than that in the LEN or ASA group, and 48 h after the drug treatment, the apoptosis induction rate of MM1.S and RPMI-8226 cells in the combination group was significantly higher than that in the single drug LEN or ASA group. Compared with LEN or ASA monotherapy group, the ratio of MM1.S and RPMI-8226 cell arrest in G1 phase was significantly increased in ASA combined with ASA group. After 48 h of treatment, compared with LEN or ASA group, the ratio of MM1.S and RPMI-8226 cell arrest was significantly higher than that of LEN or ASA group. In combination, the expression of VEGF protein in MM1.S and RPMI-8226 cells was significantly down-regulated. The expression of 尾 -catenin protein and its downstream cyclin D1 protein were up-regulated in the nucleus of MM1.S and RPMI-8226 cells after 72 h of treatment with en, and the phosphorylation of 尾 -catenin in the cytoplasm was induced by ASA. The expression of 尾 -catenin protein in the nucleus decreased in the downstream of cyclin D1 protein. Conclusion: 1, aspirin and renalidomide have synergistic effect on inhibiting the proliferation of myeloma cells. Aspirin plays a synergistic role in inhibiting myeloma by down-regulating the expression of VEGF in myeloma cell line. Aspirin can promote the proliferation of myeloma cells by promoting the proliferation of myeloma cells. Cytoplasmic 尾 -catenin phosphorylation, In order to prevent 尾-catenin into the nucleus to play the role of chemosensitization of leinadamide.
【學位授予單位】：南昌大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：R733.3

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本文編號：1951018