本文選題：斑蝥素酸鎂　切入點：抗癌活性　出處：《遵義醫(yī)學(xué)院》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的:比較自主合成新藥──斑蝥素酸鎂與斑蝥素、斑蝥素酸鈉在體外的抗癌活性,篩選出抗癌活性最好的藥物,并初步探討其抗癌作用機(jī)制。方法:1.利用CCK-8(Cell Counting Kit-8)法檢測不同濃度的斑蝥素酸鎂、斑蝥素、斑蝥素酸鈉在體外對人胃癌細(xì)胞MKN-28、SGC-7901、BGC-823增殖的影響;2.倒置相差顯微鏡下觀察不同濃度的斑蝥素酸鎂、斑蝥素、斑蝥素酸鈉作用后三種胃癌細(xì)胞形態(tài)的變化;3.以人正常胃粘膜細(xì)胞GES-1為對照,評價斑蝥素酸鎂的細(xì)胞毒性作用4.利用透射電鏡觀察斑蝥素酸鎂作用后人胃癌細(xì)胞BGC-823超微結(jié)構(gòu)的變化;5.流式細(xì)胞技術(shù)檢測斑蝥素酸鎂對人胃癌細(xì)胞BGC-823的細(xì)胞周期和細(xì)胞凋亡的影響;6.Western-Blot蛋白印跡法檢測斑蝥素酸鎂作用后人胃癌細(xì)胞BGC-823中周期素依賴性蛋白激酶1(CDK1)、細(xì)胞周期蛋白B1(Cyclin B1)蛋白的表達(dá)水平。結(jié)果:1.斑蝥素酸鎂、斑蝥素、斑蝥素酸鈉對MKN-28、SGC-7901、BGC-823三株胃癌細(xì)胞增殖均有明顯抑制作用,呈劑量-效應(yīng)關(guān)系(P0.05);斑蝥素酸鎂的抑制作用優(yōu)于斑蝥素和斑蝥素酸鈉;2.倒置相差顯微鏡觀察顯示:斑蝥素酸鎂、斑蝥素、斑蝥素酸鈉作用24h后,胃癌細(xì)胞皺縮、變圓,部分細(xì)胞破碎;3.斑蝥素酸鎂在0.52μmol/L時,已經(jīng)對胃癌細(xì)胞BGC-823表現(xiàn)出明顯的抑制作用,而對于人正常胃粘膜細(xì)胞GES-1,在1.04μmol/L時抑制率仍然很低。在相同的藥物濃度作用下,斑蝥素酸鎂對胃癌細(xì)胞BGC-823增殖的抑制作用明顯強(qiáng)于人正常胃粘膜細(xì)胞GES-1;其半數(shù)抑制濃度IC50分別為4.868μmol/L、9.645μmol/L。4.透射電鏡:半數(shù)抑制濃度的斑蝥素酸鎂作用于BGC-823細(xì)胞24h后,觀察到細(xì)胞核異形、裂解,內(nèi)質(zhì)網(wǎng)及線粒體明顯腫脹;5.碘化丙錠(propidiumiodide,PI)單染色檢測結(jié)果顯示:斑蝥素酸鎂作用于胃癌細(xì)胞BGC-823后,G0/G1期細(xì)胞比率減少,S期基本不變,G2/M期細(xì)胞增加,表現(xiàn)為G2/M期阻滯(P0.05);膜連蛋白-異硫氰酸熒光素(Annexin V-FITC/PI)雙染流式細(xì)胞術(shù)檢測結(jié)果:隨著斑蝥素酸鎂濃度增加,BGC-823細(xì)胞的凋亡率也逐漸升高(P0.05);6.Western-Blot結(jié)果顯示:斑蝥素酸鎂作用24h后,BGC-823細(xì)胞中CDK1、CyclinB1蛋白表達(dá)水平顯著降低(**P0.05,##P0.05)。結(jié)論:1.斑蝥素酸鎂對人胃癌細(xì)胞的抑制作用優(yōu)于斑蝥素和斑蝥素酸鈉;2.斑蝥素酸鎂能將胃癌細(xì)胞BGC-823阻滯在G2/M期,并誘導(dǎo)BGC-823細(xì)胞發(fā)生凋亡,其機(jī)制可能與下調(diào)CDK1、Cyclin B1蛋白的表達(dá)有關(guān);3.斑蝥素酸鎂的抗癌細(xì)胞毒性具有一定程度的細(xì)胞選擇性和敏感性。
[Abstract]:Objective: to compare the anticancer activity of magnesium cantharidate with that of cantharidin, sodium cantharidin in vitro, and to screen out the drugs with the best anticancer activity. CCK-8(Cell Counting Kit-8 method was used to detect different concentrations of magnesium cantharidate and cantharidin. Effect of Cantharidin Sodium on Proliferation of Human gastric Cancer Cell Line MKN-28 SGC-7901 BGC-823 in Vitro. Morphological changes of three gastric cancer cells after treatment with sodium cantharidin 3. GES-1 of normal gastric mucosal cells was used as control. To evaluate the cytotoxicity of magnesium cantharidin 4. To observe the ultrastructural changes of BGC-823 in human gastric cancer cells after treated with magnesium cantharidin by transmission electron microscope. Flow cytometry was used to detect the cell cycle of Cantharidin magnesium on human gastric cancer cell line BGC-823. 6. Western-Blot Western blot was used to detect the expression of cyclin dependent protein kinase (CDK1) and cyclin B1 (cyclin B1) protein in human gastric cancer cell line BGC-823 treated with magnesium cantharidate. Cantharidin and sodium cantharidate inhibited the proliferation of MKN-28 SGC-7901 and BGC-823 gastric cancer cells in a dose-effect manner, and the inhibitory effects of magnesium cantharidate were superior to those of cantharidin and sodium cantharidate. Cantharidin, sodium cantharidin, after 24 hours, the gastric cancer cells shrinked and became round, and some of the cells were broken. Magnesium cantharidin has shown obvious inhibitory effect on the BGC-823 of gastric cancer cells at 0.52 渭 mol/L. The inhibitory rate of GES-1 on normal gastric mucosal cells was still very low at 1.04 渭 mol/L. The inhibitory effect of magnesium cantharidate on BGC-823 proliferation of gastric cancer cells was significantly stronger than that on normal gastric mucosal cells GES-1, and its half inhibitory concentration (IC50) was 4.868 渭 mol / L, 9.645 渭 mol / L 路L. 4. Transmission electron microscope showed that magnesium cantharidate at half inhibitory concentration could inhibit the proliferation of BGC-823 cells for 24 hours. The results of single staining for propidiumiodide iodide (Pi) showed that the ratio of G _ 0 / G _ 1 phase to G _ 0 / G _ 1 phase of gastric cancer cells increased after treated with magnesium cantharidate, and the ratio of G _ 0 / G _ 1 phase to G _ 2 / M phase increased significantly after treatment with Cantharidin. The results of flow cytometry showed that the apoptosis rate of BGC-823 cells increased with the increase of magnesium cantharidate concentration, and the results of Western-Blot showed that: the effect of Cantharidin on magnesium Cantharidin was found in BGC-823 cells with the increase of the concentration of magnesium cantharidin. The results of Western-Blot showed that: the effect of Cantharidin on magnesium Cantharidin was determined by flow cytometry, and the apoptosis rate of BGC-823 cells increased with the increase of concentration of magnesium cantharidate. After 24 hours, the expression of CDK1 and CyclinB1 in BGC-823 cells decreased significantly. Conclusion: 1. The inhibitory effect of magnesium cantharidin on human gastric cancer cells is superior to that of cantharidin and sodium cantharidate. Magnesium cantharidin can block the BGC-823 of gastric cancer cells at G 2 / M stage. The mechanism may be related to the down-regulation of CDK1 Cyclin B1 protein expression. Cantharidin magnesium has a certain degree of cell selectivity and sensitivity to anticancer cytotoxicity.
【學(xué)位授予單位】：遵義醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R735.2

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