本文選題：雙氫青蒿素 + 紫杉醇��； 參考：《南華大學(xué)》2016年碩士論文

【摘要】：目的:以人胃癌MGC-803細(xì)胞為研究對(duì)象,觀察雙氫青蒿素與紫杉醇對(duì)該細(xì)胞增殖和凋亡的影響,以初步探討兩藥聯(lián)合是否能起到協(xié)同作用,從而為胃癌的化療提供新思路。方法:體外培養(yǎng)MGC-803細(xì)胞,采用CCK-8法于24h分別檢測(cè)不同濃度雙氫青蒿素、紫杉醇及兩藥聯(lián)合對(duì)細(xì)胞增殖的影響,并用CHOU-Talalay法評(píng)估兩藥聯(lián)合的效果;用流式細(xì)胞儀分析各藥物組對(duì)細(xì)胞周期阻滯的影響;用AO/EB染色劑處理細(xì)胞后,在熒光顯微鏡下觀察各藥物處理組作用MGC-803細(xì)胞后的凋亡情況;用逆轉(zhuǎn)錄聚合酶鏈反應(yīng)法和免疫蛋白印跡法檢測(cè)對(duì)人胃癌MGC-803細(xì)胞的Bcl-2、Bax和Caspase-3表達(dá)的影響。結(jié)果:1、雙氫青蒿素(191.082、955.410、4777.048、23885.240、119426.202ng/ml)、紫杉醇(2.4、12、60、300、1500ng/ml)及兩藥聯(lián)合(191.082+2.4、955.410+12、4777.048+60、23885.240+300、119426.202+1500ng/ml)對(duì)人胃癌MGC-803細(xì)胞均具有增殖抑制作用并呈濃度依賴(lài)(p0.05);聯(lián)合用藥組的增殖抑制作用優(yōu)于單一用藥組(p0.05),其中兩藥小劑量聯(lián)合用藥時(shí)呈協(xié)同作用,大劑量聯(lián)合用藥時(shí)呈拮抗作用。2、通過(guò)流式細(xì)胞儀對(duì)藥物處理后細(xì)胞的周期檢測(cè),我們發(fā)現(xiàn)雙氫青蒿素(955.410ng/ml)作用后出現(xiàn)G0/G1期、S期細(xì)胞阻滯(p0.001),紫杉醇(12ng/ml)作用后出現(xiàn)G2/M期細(xì)胞阻滯(p0.001),兩藥聯(lián)合(DHA955.410ng/ml+PTX12ng/ml)表現(xiàn)出S期、G2/M期阻滯(p0.001);聯(lián)合用藥組相比于紫杉醇用藥組有G2/M期細(xì)胞阻滯減少(p0.001)。3、熒光顯微鏡觀察發(fā)現(xiàn),雙氫青蒿素(955.410ng/ml)、紫杉醇(12ng/ml)處理MGC-803細(xì)胞24h后出現(xiàn)較多橙黃及橙紅色細(xì)胞,相比于綠色細(xì)胞而言,體積縮小、胞質(zhì)濃縮,具有凋亡細(xì)胞的明顯特征。同樣條件下,聯(lián)合組的凋亡細(xì)胞數(shù)較單藥組明顯增多,且晚期凋亡細(xì)胞數(shù)目居多。4、雙氫青蒿素(955.410ng/ml)及紫杉醇(12ng/ml)作用于人胃癌MGC-803細(xì)胞后均能促進(jìn)Bax、Caspase-3的表達(dá),抑制Bcl-2的表達(dá),而兩藥聯(lián)合(DHA955.410ng/ml+PTX12ng/ml)表現(xiàn)出更明顯的效果,差異均有統(tǒng)計(jì)學(xué)意義(p0.05)。結(jié)論:1、雙氫青蒿素、紫杉醇對(duì)人胃癌MGC-803細(xì)胞的增殖抑制呈濃度依賴(lài);2、兩藥聯(lián)合對(duì)MGC-803細(xì)胞有增殖抑制作用,它們合用時(shí)小劑量是協(xié)同作用,而大劑量則為拮抗作用;3、兩藥均可通過(guò)增加Bax、Caspase-3的表達(dá)及減少Bcl-2的表達(dá)來(lái)誘導(dǎo)細(xì)胞凋亡。
[Abstract]:Objective: to observe the effects of dihydroartemisinin and paclitaxel on the proliferation and apoptosis of human gastric cancer cell line MGC-803 in order to explore whether the combination of the two drugs can play a synergistic effect and provide a new way for the chemotherapy of gastric cancer. Methods: MGC-803 cells were cultured in vitro. The effects of dihydroartemisinin, paclitaxel and the combination of two drugs on the proliferation of MGC-803 cells were detected by CCK-8 method at 24 hours, and the effects of the two drugs were evaluated by CHOU-Talalay method. The cell cycle arrest was analyzed by flow cytometry, and the apoptosis of MGC-803 cells treated with AO-EB staining was observed under fluorescence microscope. Reverse transcriptase polymerase chain reaction (RT-PCR) and Western blotting were used to detect the expression of Bcl-2P Bax and Caspase-3 in human gastric cancer MGC-803 cells. Results: 1, 191.082955.41010, 4777.048, 23885.240, 19426.2026 ng / ml, paclitaxel 2.4126060300 / 1500ng / ml) and two drugs combined, 191.082 2.4955.410, 121.082 2.4955.410, 121.082 2.4955.410, 121.082 2.4955.410, 121.082 2.4955.410, 23887.048 60,23887.048 60,19426.202 1500ng / ml) all had inhibitory effects on the proliferation of human gastric cancer MGC-803 cells in a concentration-dependent manner, and the inhibitory effect of the combined treatment group was superior to that of the single drug group (p0.05N), two of which were found to be in a dose-dependent manner. There was a synergistic effect when the drug was combined with small dosage. The cell cycle after drug treatment was detected by flow cytometry. We found that after the treatment of dihydroartemisinin (955.410ng / ml), the cell block of G _ 0 / G _ 1 phase in S phase was found, and that of paclitaxel 12ng / ml group showed G _ 2 / M phase cell block P _ (0.001), and the combination of two drugs, DHA 955.410ng / ml PTX _ (12ng / ml), showed S phase G _ 2M phase block (P _ 0.001), compared with paclitaxel group (P _ 0.001). There was a decrease in G _ 2 / M phase arrest, P 0.001 ~ (-1) ~ 3, and fluorescence microscopy showed that, MGC-803 cells treated with dihydroartemisinin 955.410 ng / ml, paclitaxel 12ng / ml for 24 h showed more orange and orange red cells. Compared with green cells, the cells were smaller in volume and concentrated in cytoplasm and had the obvious characteristics of apoptotic cells. Under the same conditions, the number of apoptotic cells in the combination group was significantly higher than that in the single drug group, and the number of apoptotic cells in the late stage was mainly .4.The expression of BaxCaspase-3 was inhibited and the expression of BaxCaspase-3 was inhibited by the addition of dihydroartemisinin 955.410ng / ml and taxol 12ng / ml to human gastric cancer MGC-803 cells. The combination of DHA 955.410ng / ml PTX 12ng / ml showed a more obvious effect, and the difference was statistically significant (P 0.05). Conclusion the inhibition of proliferation of human gastric cancer MGC-803 cells by 1: 1, dihydroartemisinin and paclitaxel is concentration-dependent. The combination of the two drugs can inhibit the proliferation of MGC-803 cells. However, the antagonistic effect of high dose was antagonistic. Both drugs could induce apoptosis by increasing the expression of Baxton-Caspase-3 and decreasing the expression of Bcl-2.
【學(xué)位授予單位】：南華大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類(lèi)號(hào)】：R735.2

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