【摘要】：目的:研究?jī)煞N新型拓?fù)涿敢种苿?-甲基- 11- (4-二甲基氨基芐烯)-7,8,9,10-四氫-1-氫吲哚[1,2-b]吲噠唑三氟甲磺酸鹽(受試物1)和1-甲基- 5- (4-二甲氨基芐烯)-5H-茚[1,2-b]吡啶三氟甲磺酸鹽(受試物2)體外抗腫瘤活性,并初步探討其作用機(jī)制。 方法:采用MTT法檢測(cè)兩種受試物對(duì)人腫瘤細(xì)胞增殖的抑制作用;質(zhì)粒pBR322解旋反應(yīng)考察受試物對(duì)TopoⅠ催化活性的影響,Western blot法檢測(cè)細(xì)胞中TopoⅠ和TopoⅡ的蛋白表達(dá)變化;Hoechst 33258熒光染色、AO熒光染色、單細(xì)胞凝膠電泳和流式細(xì)胞儀等方法觀察兩種受試物對(duì)HEp-2細(xì)胞凋亡和細(xì)胞周期的影響;Western blot法分析受試物導(dǎo)致HEp-2細(xì)胞死亡的機(jī)制。 結(jié)果:MTT法的結(jié)果顯示受試物1和受試物2對(duì)9種人腫瘤細(xì)胞株均有顯著的抗腫瘤活性,并呈現(xiàn)濃度依賴性;受試物1和受試物2分別在8μmol·L-1和4μmol·L-1時(shí)可抑制TopoⅠ介導(dǎo)的質(zhì)粒DNA超螺旋的解旋,并且兩種受試物都可下調(diào)細(xì)胞中TopoⅠ、Ⅱ的蛋白表達(dá);Hoechst熒光染色表明受試物可誘導(dǎo)HEp-2細(xì)胞凋亡,AO染色也得到相同的結(jié)果;單細(xì)胞凝膠電泳檢測(cè)出兩種受試物導(dǎo)致DNA損傷;流式細(xì)胞儀檢測(cè)的結(jié)果表明處理組的細(xì)胞凋亡率高于對(duì)照組,同時(shí)顯示,受試物1使HEp-2細(xì)胞周期在S期和G2/M期發(fā)生阻滯,而受試物2使其阻滯在G1期;Western blot檢測(cè)細(xì)胞凋亡及細(xì)胞周期相關(guān)蛋白,結(jié)果顯示促凋亡蛋白Bid、Bax和JNK表達(dá)增加,抗凋亡蛋白Bcl-2表達(dá)減少,細(xì)胞周期蛋白cyclin B1和cyclin D1表達(dá)減少 結(jié)論:本文實(shí)驗(yàn)表明,6-甲基- 11- (4-二甲基氨基芐烯)-7,8,9,10-四氫- 1-氫吲哚[1,2-b]吲噠唑三氟甲磺酸鹽和1-甲基-5-(4-二甲氨基芐烯)-5H-茚[1,2-b]吡啶三氟甲磺酸鹽可以抑制HEp-2細(xì)胞的生長(zhǎng),這可能與以下機(jī)制有關(guān):①通過(guò)抑制TopoⅠ的催化活性并下調(diào)細(xì)胞中TopoⅠ和TopoⅡ的表達(dá);②調(diào)控凋亡相關(guān)基因的表達(dá),誘導(dǎo)其凋亡;③造成腫瘤細(xì)胞周期紊亂,促進(jìn)凋亡。
[Abstract]:Objective: to study two novel topological enzyme inhibitors, 6-methyl-11- (4-dimethylaminobenzene) -7-, 9 / 10-tetrahydro-1-hydroindole-[ 1] indazolate-trifluoromethyl sulfonate (test 1) and 1-methyl-5- (4-dimethylaminobenzene) -5H-indene [1t2-b] pyridine trifluoromethyl sulfonate (subject) 2) Antitumor activity in vitro, The mechanism of its action was also discussed. Methods: MTT assay was used to detect the inhibitory effect of two kinds of samples on the proliferation of human tumor cells, and the effect of plasmid pBR322 unspin reaction on the catalytic activity of Topo 鈪，

本文編號(hào)：2334910