目的 探討異戊烯基香豆素類化合物對(duì)A549肺癌細(xì)胞增殖的影響，并對(duì)其構(gòu)效關(guān)系及作用機(jī)制進(jìn)行初步探討。方法 將人肺癌細(xì)胞A549進(jìn)行體外培養(yǎng)，采用不同濃度異戊烯基香豆素類化合物甘草香豆素（GCM）、甘草酚（GC）、補(bǔ)骨脂定（PSO）、香豆雌酚（COM）、7-去甲基軟木花椒素（DE）、7, 2', 4'-trihydroxy-5-methxy-3-penylcoumarin（TMP）和蛇床子素（OST）分別處理肺癌A549細(xì)胞36、72 h后，通過(guò)結(jié)晶紫染色法檢測(cè)細(xì)胞增殖活性；采用蛋白免疫印跡法（Western blotting）檢測(cè)細(xì)胞中TOPK、C-PARP、Bcl-2、Bax、p62、LC3B蛋白表達(dá)水平。結(jié)果 受試化合物對(duì)A549細(xì)胞增殖的抑制作用存在濃度和時(shí)間相關(guān)性；10 μmol/L的GCM、GC、PSO、COM、DE、TMP組Bcl-2表達(dá)均下降（P＜0.05）；A549細(xì)胞經(jīng)10 μmol/L GCM、GC、PSO處理后，TOPK的表達(dá)顯著降低，C-PARP、LC3 Ⅱ、p62的表達(dá)顯著升高（P＜0.05）。結(jié)論 異戊烯基香豆素類化合物GCM、GC、PSO可顯著抑制A549細(xì)胞的增殖活性，且其抗增殖活性可能與香豆素母核苯環(huán)上的異戊烯基結(jié)構(gòu)取代和母核C-3位上苯環(huán)的取代及其環(huán)上游離的羥基有關(guān)；其抗A549細(xì)胞增殖的作用機(jī)制可能與促進(jìn)細(xì)胞凋亡，誘導(dǎo)細(xì)胞發(fā)生不完全自噬并抑制自噬流有關(guān)。

Objective To explore the influence of prenylated coumarins on the proliferation of lung cancer cells and preliminary explored on its structure-activity relationship and inhibitory mechanism.Methods The human lung cancer cell A549 was cultured in vitro,the cells were treated with different concentrations of Glycycoumarin (GCM),Glycyrol (GC),Psoralidin (PSO),Coumestrol (COM),Demethylsuberosin (DE),7,2',4'-trihydroxy-5-methxy-3-penylcoumarin (TMP),Osthole (OST) for 36 and 72 h.Crystal violet staining was used to detect the activity on the proliferation of A549 cells.Western blotting was used to detect the protein expressions of TOPK,C-PARP,Bcl-2,Bax,p62 and LC3B in the cells.Results The test compounds inhibited the proliferation of A549 cells in a concentration and time-dependent manner.Bcl-2 expression in 10 μmol/L GCM,GC,PSO,COM,DE,TMP groups were decreased (P<0.05).After treatment with 10 μmol/L GCM,GC and PSO,the expression of TOPK in A549 lung cancer cells was significantly decreased,while the expression of C-PARP,LC3II and p62 were significantly increased (P<0.05).Conclusion Prenylated coumarins compound of GCM,GC,PSO can significantly inhibit proliferation activity of A549 cells,and its anti-proliferative activity may be substituted with isoprene structures on coumarin phenylring and mother core.The substitution of the C-3-bit benzene ring is related to the hydroxyl group across the ring,the mechanism of the anti-A549 cell proliferation may be related to the promotion of apoptosis,inducing cells induced by incomplete autophagy and inhibiting autophagy stream.

R285

貴州省科技計(jì)劃項(xiàng)目(黔科合支撐[2020]4Y118號(hào));畢節(jié)市科技局重點(diǎn)實(shí)驗(yàn)室項(xiàng)目(畢科合字[2019]1號(hào))