本文選題：紫草素 + NB4細(xì)胞��； 參考：《重慶醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:研究紫草素對(duì)人急性早幼粒細(xì)胞白血病NB4細(xì)胞增殖與凋亡的影響及其相關(guān)作用機(jī)制。方法:1.以NB4細(xì)胞株作為研究對(duì)象,用梯度濃度的紫草素分別作用于NB4細(xì)胞不同時(shí)間后,CCK-8實(shí)驗(yàn)測(cè)定細(xì)胞增殖情況;2.以0.3μmol/L紫草素作用于NB4細(xì)胞,培養(yǎng)1 d后,通過(guò)流式細(xì)胞術(shù)檢測(cè)細(xì)胞周期的改變和凋亡的改變;3.經(jīng)Hochest 33342實(shí)驗(yàn),通過(guò)熒光顯微鏡檢測(cè)NB4細(xì)胞的胞核的形態(tài)學(xué)改變;4.Western blot檢測(cè)細(xì)胞c-Myc,PARP,Caspase 3,Cleaved PARP,Cleaved Caspase 3,ERK,p-ERK,JNK,p-JNK,p38MAPK及p-p38MAPK蛋白表達(dá)情況;5.構(gòu)建NB4細(xì)胞裸鼠移植瘤模型,通過(guò)描繪腫瘤生長(zhǎng)曲線分析紫草素對(duì)裸鼠腫瘤發(fā)展情況的作用;6.通過(guò)免疫組化技術(shù)檢測(cè)紫草素對(duì)抑制瘤組織中增殖指標(biāo)c-Myc表達(dá)的影響。結(jié)果:1.NB4細(xì)胞經(jīng)紫草素處理后,細(xì)胞活力受到抑制(P0.01),并且具有時(shí)間依賴性及濃度依賴性;2.流式細(xì)胞術(shù)檢測(cè)到NB4細(xì)胞經(jīng)紫草素處理后,G1期細(xì)胞比例上升(P0.01),G2期和S期細(xì)胞比例下降(P0.01);細(xì)胞凋亡率明顯增加(P0.01);3.紫草素處理NB4后,經(jīng)Hochest 33342著色,在熒光顯微鏡下發(fā)現(xiàn)細(xì)胞表現(xiàn)出核固縮,染色質(zhì)聚集凝集,核破碎等現(xiàn)象;4.紫草素作用細(xì)胞1 d后,Western blot檢測(cè)到凋亡相關(guān)蛋白Cleaved PARP,Cleaved Caspase 3表達(dá)明顯增多(P0.01),p-JNK和p-p38MAPK蛋白水平明顯升高(P0.01),p-ERK和c-Myc蛋白表達(dá)水平明顯降低(P0.05);5.紫草素能夠顯著抑制移植瘤生長(zhǎng)(P0.05);6.免疫組化結(jié)果顯示,紫草素處理組增殖蛋白c-Myc表達(dá)與對(duì)照組相比顯著降低(P0.05)。結(jié)論:紫草素可能通過(guò)調(diào)節(jié)MAPK通路和c-Myc表達(dá)水平來(lái)抑制NB4細(xì)胞增殖誘導(dǎo)其凋亡,并可抑制NB4細(xì)胞裸鼠移植瘤生長(zhǎng)。
[Abstract]:Aim: to study the effect of porphyrin on proliferation and apoptosis of human acute promyelocytic leukemia (NB4) cells and its related mechanism.Method 1: 1.NB4 cell line was used as the research object. The proliferation of NB4 cells was measured by CCK-8 assay after treated with the gradient concentration of porphyrin for different time.NB4 cells were treated with 0.3 渭 mol/L porphyrin for 1 day. The cell cycle and apoptosis were detected by flow cytometry.The morphological changes of the nucleus of NB4 cells were detected by Hochest 33342 assay. 4. Western blot was used to detect the expression of c-Myccine paspase 3 and Cleaved PARPASE p38 MAPK and p-p38MAPK protein in NB4 cells.The model of transplanted tumor in nude mice with NB4 cells was constructed, and the effect of porphyrin on tumor development in nude mice was analyzed by depicting tumor growth curve.The effect of Shikonin on the expression of c-Myc in tumor tissue was detected by immunohistochemical technique.Results: 1. The cell viability of Nb4 cells was inhibited after treated with porphyrin, and the cell viability was time-dependent and concentration-dependent.Flow cytometry (FCM) showed that the proportion of NB4 cells in G _ 1 phase increased after treatment with Shikonin, and the proportion of P0.01G _ 2 and S phase cells decreased (P _ (0.01)), and the apoptosis rate increased significantly (P _ (0.01)).After NB4 was treated with porphyrin, the cells were stained with Hochest 33342 and the cells showed nuclear pyknosis, chromatin aggregation and nuclear fragmentation under fluorescence microscope.One day after treatment with porphyrin, the expression of apoptosis-related protein Cleaved, p0.01, p-JNK and p-p38MAPK was significantly increased by Western blot. The expression level of P0.01PERK and c-Myc decreased significantly.Shikonin could significantly inhibit the growth of transplanted tumor.Immunohistochemical results showed that the expression of proliferating protein (c-Myc) in the treated group was significantly lower than that in the control group (P 0.05).Conclusion: Shikonin may inhibit the proliferation and apoptosis of NB4 cells by regulating the MAPK pathway and the expression of c-Myc, and inhibit the growth of NB4 xenografts in nude mice.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R733.7

【參考文獻(xiàn)】

相關(guān)期刊論文 前5條

1 劉華鋼;劉麗敏;黃慧學(xué);楊斌;梁秋云;;抗腫瘤中藥動(dòng)物模型及機(jī)制研究進(jìn)展[J];現(xiàn)代生物醫(yī)學(xué)進(jìn)展;2008年07期

2 李麗霞;湯永民;;人類血液系腫瘤動(dòng)物模型的建立及其靶向治療研究進(jìn)展[J];實(shí)用腫瘤雜志;2007年03期

3 張奇;白曉東;付小兵;;p38MAPK信號(hào)通路研究進(jìn)展[J];感染 炎癥 修復(fù);2005年02期

4 ;MAPK signal pathways in the regulation of cell proliferation in mammalian cells[J];Cell Research;2002年01期

5 郭喜平;張興義;張署東;劉文學(xué);王榮有;馬赤;田榮閣;趙守先;;紫星口服液治療晚期肺癌的臨床觀察[J];中西醫(yī)結(jié)合雜志;1991年10期

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