發(fā)布時間：2018-04-08 17:38

  本文選題：白血病　切入點：耐藥　出處：《延邊大學》2017年碩士論文

【摘要】：目的:探討CD44在K562細胞及阿霉素耐藥細胞株K562/ADR中的表達情況,闡明CD44與K562細胞耐藥之間的相關性,探索逆轉(zhuǎn)白血病耐藥的途徑。方法:1.利用實時熒光定量PCR法,檢測K562細胞及K562/ADR細胞中CD44的mRNA表達水平;2.用電轉(zhuǎn)染法,制備高表達CD44的K562細胞(K562/CD44細胞);3.用倒置熒光顯微鏡下觀察法、實時熒光定量PCR法,檢測K562和K562/CD44細胞的CD44標記綠色熒光蛋白、CD44的mRNA表達水平,確認轉(zhuǎn)染成功與否;4.WST-1細胞增殖實驗,檢測K562細胞的阿霉素IC50,選擇適宜的阿霉素藥物濃度和作用時間,并對比K562和K562/CD44細胞在阿霉素相同濃度和相同時間作用下的存活率。結(jié)果:1.實時熒光定量PCR檢測結(jié)果:與K562細胞比較K562/ADR細胞中的CD44 mRNA表達水平顯著上升(P0.01);2.倒置熒光顯微鏡觀察結(jié)果顯示:CD44高表達載體轉(zhuǎn)染后的細胞具有綠色熒光蛋白的高表達;3.實時熒光定量PCR檢測結(jié)果:與K562細胞相比較K562/CD44細胞中的CD44 mRNA表達顯著上調(diào)(P0.01);4.WST-1 細胞增殖實驗檢測結(jié)果:(1)0.25mg/L、0.5mg/L、1.Omg/L、1.5mg/L和2.0mg/L的阿霉素作用于K562細胞24h、48h、72h均顯著抑制細胞增殖,并呈時間和濃度依賴性,1.0mg/L、1.5mg/L和2.0mg/L的阿霉素抑制細胞增殖效應無顯著差異(P0.05),K562細胞的阿霉素IC50為0.7376mg/L;(2)與K562細胞比較,K562/CD44細胞在1.0mg/L阿霉素作用48h時的存活率顯著上升(P0.01)。結(jié)論:1.CD44過表達的K562細胞對阿霉素的敏感性減弱;2.CD44的過表達可能參與了 K562細胞的耐藥機制。
[Abstract]:Objective: to investigate the expression of CD44 in K562 cells and doxorubicin resistant cell line K562/ADR, to elucidate the correlation between CD44 and drug resistance of K562 cells, and to explore the way to reverse the drug resistance of leukemia.Method 1: 1.The mRNA expression of CD44 in K562 cells and K562/ADR cells was detected by real-time fluorescence quantitative PCR assay.K562 cells with high expression of CD44 were prepared by electrotransfection.The expression of CD44 labeled green fluorescent protein CD44 (CD44) in K562 and K562/CD44 cells was detected by reverse fluorescence microscope and real-time fluorescence quantitative PCR, and the proliferation of K562 and K562/CD44 cells was confirmed by 4.WST-1 cell proliferation.The doxorubicin IC50 of K562 cells was detected, and the optimal concentration and time of doxorubicin treatment were selected, and the survival rate of K562 and K562/CD44 cells was compared under the same concentration and time of doxorubicin.The result is 1: 1.Results of real-time fluorescence quantitative PCR: compared with K562 cells, the expression of CD44 mRNA in K562/ADR cells increased significantly (P 0.01).The results of inverted fluorescence microscope showed that the cells transfected with the high expression vector of 1% CD44 had high expression of green fluorescent protein (GFP).Conclusion the over-expression of CD44 in K562 cells may be involved in the mechanism of drug resistance in K562 cells.
【學位授予單位】：延邊大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R733.7

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