【摘要】：目的 在本論文中,通過(guò)MTT等體外實(shí)驗(yàn)觀察核苷類化合物N對(duì)多個(gè)腫瘤細(xì)胞系的抗腫瘤活性,以及初步探討該核苷化合物可能具有的抗腫瘤作用機(jī)制,從而為開(kāi)發(fā)新的抗腫瘤化療藥物或發(fā)現(xiàn)先導(dǎo)化合物奠定初步的理論基礎(chǔ)。 方法 1.通過(guò)常規(guī)細(xì)胞毒性實(shí)驗(yàn)MTT法,觀察核苷類化合物N在體外試驗(yàn)中對(duì)細(xì)胞株JEG-3、HepG2、細(xì)胞系HL-60的細(xì)胞毒活性； 2.JEG-3細(xì)胞、HL-60細(xì)胞體外經(jīng)核苷類化合物N (0.1μM、0.2μM)處理24小時(shí)后,細(xì)胞經(jīng)4%多聚甲醛固定及核染料Hoechst33258染色后,置熒光倒置顯微鏡下觀察核苷類化合物N誘導(dǎo)細(xì)胞凋亡的形態(tài)學(xué)變化； 3.采用western blot技術(shù),觀察HL-60細(xì)胞經(jīng)核苷類化合物N (0.1μM、0.2μM)處理后,核苷類化合物N對(duì)HL-60細(xì)胞凋亡有關(guān)蛋白caspase-3和AKT表達(dá)的調(diào)節(jié)作用。 結(jié)果 1.核苷類化合物N在體外能抑制HL-60細(xì)胞和JEG-3細(xì)胞的正常增殖,并表現(xiàn)出了明顯的量效依賴關(guān)系； 2.不同濃度核苷類化合物N (0.1μM,0.2μM)處理HL-60細(xì)胞和JEG-3細(xì)胞24小時(shí)后,細(xì)胞經(jīng)固定和Hoechst33258染色,熒光倒置顯微鏡觀察發(fā)現(xiàn),細(xì)胞細(xì)胞核形態(tài)發(fā)生明顯改變,細(xì)胞核呈現(xiàn)染色不均,出現(xiàn)核切跡,核濃縮聚集,甚至細(xì)胞核碎裂。核苷類化合物N處理后細(xì)胞形態(tài)學(xué)表現(xiàn)出典型的凋亡改變； 3.體外核苷類化合物N (0.1μM、0.2μM)體外處理HL-60細(xì)胞24小時(shí)后,應(yīng)用western blot技術(shù),實(shí)驗(yàn)結(jié)果發(fā)現(xiàn),核苷類化合物N可以明顯下調(diào)HL-60細(xì)胞抗凋亡蛋白AKT的表達(dá)；經(jīng)核苷類化合物N作用后,HL-60細(xì)胞出現(xiàn)caspase-3的裂解片段(17KD)。 結(jié)論 合成的核苷類化合物N在體外可以劑量依賴性地抑制多個(gè)腫瘤細(xì)胞株的生長(zhǎng)繁殖,具有明顯的細(xì)胞毒活性；核苷化合物N的抗腫瘤活性可能和其能調(diào)節(jié)腫瘤細(xì)胞凋亡相關(guān)蛋白表達(dá),進(jìn)而誘導(dǎo)腫瘤細(xì)胞凋亡有關(guān)�；衔颪在體外試驗(yàn)中表現(xiàn)出了顯著的抗腫瘤活性,該化合物的抗腫瘤活性和其具體抗腫瘤作用機(jī)制值得進(jìn)一步深入開(kāi)發(fā)研究。
[Abstract]:Objective to observe the antitumor activity of nucleoside compound N on several tumor cell lines in vitro by MTT and to explore the possible mechanism of the antitumor effect of the nucleoside compound. The results provide a theoretical basis for the development of new anti-tumor chemotherapeutic drugs or the discovery of lead compounds. Method 1. The cytotoxic activity of nucleoside compound N to HL-60 cell line JEG-3,HepG2, cell line was observed by conventional cytotoxicity test (MTT) in vitro. 2.JEG-3 cells and HL-60 cells were treated with N (0.1 渭 M) 0.2 渭 M for 24 hours in vitro. The cells were fixed with 4% paraformaldehyde and stained with nuclear dye Hoechst33258. The morphological changes of apoptosis induced by nucleoside compound N were observed under fluorescence inverted microscope. 3. Western blot technique was used to observe the regulatory effect of nucleoside N on the expression of caspase-3 and AKT in HL-60 cells treated with nucleoside compound N (0.1 渭 M 0.2 渭 M). Result 1. Nucleoside compound N could inhibit the normal proliferation of HL-60 cells and JEG-3 cells in vitro and showed a dose-dependent relationship. 2. After treated with different concentrations of nucleoside compound N (0.1 渭 M, 0.2 渭 M) for 24 hours, HL-60 cells and JEG-3 cells were fixed and stained with Hoechst33258, and observed by fluorescence inverted microscope. The nuclei showed uneven staining, nuclear cuts, nuclear condensation and aggregation, and even nuclear fragmentation. After treatment with nucleoside compound N, the cell morphology showed typical apoptotic changes. 3. Nuclear glycosides N (0. 1 渭 M + 0. 2 渭 M) were used to treat HL-60 cells for 24 hours. The results showed that nucleoside N could significantly down-regulate the expression of anti apoptotic protein AKT in HL-60 cells. After treatment with nucleoside compound N, caspase-3 fragmentation fragment (17KD) appeared in HL-60 cells. Conclusion the synthesized nucleoside compound N can inhibit the growth and reproduction of many tumor cell lines in a dose-dependent manner in vitro and has obvious cytotoxic activity. The antineoplastic activity of nucleoside N may be related to its ability to regulate the expression of apoptosis-related proteins in tumor cells and then induce apoptosis of tumor cells. Compound N showed significant antitumor activity in vitro, and its antitumor activity and its specific antitumor mechanism are worthy of further study.
【學(xué)位授予單位】：新鄉(xiāng)醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R96

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1 余家華;李秋林;鄭榮壽;張思維;葛蓮英;曹驥;陳萬(wàn)青;;中國(guó)2009年喉惡性腫瘤發(fā)病和死亡分析[J];中國(guó)腫瘤;2013年05期

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本文編號(hào)：2390389