【摘要】：目的:探討氧化苦參堿對(duì)皮膚鱗狀細(xì)胞癌(c SCC)Colo-16細(xì)胞株凋亡及自噬的影響。方法:用不同濃度的氧化苦參堿處理Colo-16細(xì)胞株24 h后,用流式細(xì)胞儀測(cè)定不同濃度氧化苦參堿組Colo-16細(xì)胞株凋亡率,用免疫印跡(western blot)法測(cè)定不同濃度組細(xì)胞自噬相關(guān)蛋白LC3-Ⅰ、LC3-Ⅱ的表達(dá)及LC3-Ⅰ→LC3-Ⅱ轉(zhuǎn)化率。同時(shí)用western blot法檢測(cè)哺乳動(dòng)物雷帕霉素靶蛋白(m TOR)、m TOR調(diào)節(jié)相關(guān)蛋白(Raptor)及其m TORser2448、ser2481位點(diǎn)磷酸化產(chǎn)物和Raptor ser792位點(diǎn)磷酸化產(chǎn)物的表達(dá)。結(jié)果:1.000 g/L、2.000 g/L、4.000 g/L氧化苦參堿濃度組凋亡率明顯高于陰性對(duì)照組,差異有統(tǒng)計(jì)學(xué)意義(F=355.12,P0.05),不同濃度組之間凋亡率差異也有統(tǒng)計(jì)學(xué)意義(P0.05)。western blot法發(fā)現(xiàn),陰性對(duì)照組、溶劑對(duì)照組及氧化苦參堿分別為0.025 g/L、0.050 g/L、0.100 g/L、0.500 g/L、1.000 g/L、2.000 g/L各組之間LC3-Ⅰ、LC3-Ⅱ的表達(dá)及LC3-Ⅰ→LC3-Ⅱ轉(zhuǎn)化率比較,差異無(wú)統(tǒng)計(jì)學(xué)意義(F值分別為0.86,0.63,0.33,P0.05),各組別之間m TOR、Raptor蛋白及其m TORser2448、ser2481位點(diǎn)磷酸化產(chǎn)物和Raptor ser792位點(diǎn)磷酸化產(chǎn)物的表達(dá),差異均無(wú)統(tǒng)計(jì)學(xué)意義(F值分別為1.33、1.42、1.12、0.95及0.47,P0.05)。結(jié)論:氧化苦參堿可上調(diào)Colo-16細(xì)胞株的凋亡,且與其濃度正相關(guān)。氧化苦參堿對(duì)細(xì)胞自噬的水平無(wú)影響,對(duì)自噬相關(guān)調(diào)控蛋白的表達(dá)也無(wú)影響。因此,氧化苦參堿可能通過(guò)上調(diào)Colo-16細(xì)胞株的凋亡而非自噬來(lái)實(shí)現(xiàn)其抗腫瘤效應(yīng)。
[Abstract]:Objective: to investigate the effect of oxymatrine on apoptosis and autophagy of (c SCC) Colo-16 cell line. Methods: Colo-16 cells were treated with different concentrations of oxymatrine for 24 h. The apoptosis rate of Colo-16 cells in different concentrations of oxymatrine was measured by flow cytometry. The expression of autophagy-associated proteins (LC3- 鈪，

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