基于mTOR/STAT3 信号轴探讨桑色素诱导非小细胞肺癌A549 细胞自噬的机制。方法将A549 细胞分为空白组及30、60、90、120、150 μg·mL-1 桑色素组，培养24、48、72 h 后采用CCK-8 法检测细胞增 殖活性，计算细胞抑制率。将A549 细胞分为空白组及30、90、150 μg·mL-1 桑色素组，培养细胞14 d 后通过 克隆形成实验检测细胞增殖情况；培养细胞24 h 后，采用BeyoClickTM EdU-488 检测细胞增殖能力；流式细胞 术检测细胞凋亡情况；吖啶橙染色检测细胞自噬情况；透射电镜观察细胞自噬体形成情况；Western Blot 法检 测细胞中凋亡、自噬及mTOR/STAT3 信号轴相关蛋白的表达水平。将A549 细胞分为空白组、空白+氯喹 （10 μg·mL-1）组、桑色素（30、150 μg·mL-1）组、桑色素（30、150 μg·mL-1）+氯喹（10 μg·mL-1）组，干预48 h 后 采用CCK-8 法检测细胞活性， 计算细胞存活率。结果与空白组比较， 干预24 h 后60、90、120、 150 μg·mL-1 桑色素组的A549 细胞抑制率显著升高（P＜0.05，P＜0.001）；干预48、72 h 后桑色素30、60、 90、120、150 μg·mL-1 组的A549 细胞抑制率显著升高（P＜0.001）；30、90、150 μg·mL-1 桑色素组的A549 细 胞集落数及绿色荧光的增殖阳性细胞数均显著减少（P＜0.01，P＜0.001），细胞凋亡率均显著升高（P＜0.01，P＜ 0.001），cleaved-PARP 蛋白表达水平显著升高（P＜0.001）；90、150 μg·mL-1 桑色素组A549 细胞的p-P38/P38 MAPK 蛋白表达水平显著升高（P＜0.01，P＜0.001）；30、90、150 μg·mL-1 桑色素组A549 细胞中出现不同程 度的橙黄色荧光，以90、150 μg·mL-1 桑色素组的橙黄色荧光显著；150 μg·mL-1 桑色素组A549 细胞胞浆中分 别出现了自噬体和自噬溶酶体；150 μg·mL-1 桑色素组A549 细胞的LC3-Ⅱ蛋白表达明显上调（P＜0.05），90、 150 μ g · mL-1 桑色素组A549 细胞的Atg16L1-Ⅱ 蛋白表达显著上调（P ＜ 0.001）， p-mTOR/mTOR 及 p-STAT3/STAT3 蛋白表达显著下调（P＜0.001）。与桑色素（150 μg·mL-1）组比较，桑色素（150 μg·mL-1）+氯喹 （10 μg·mL-1）组的A549 细胞存活率明显升高（P＜0.05）。结论桑色素能够抑制肺癌A549 细胞的增殖，促进 其凋亡，并诱导自噬，其作用机制可能与mTOR/STAT3 信号轴有关

To investigate the mechanism of morin-induced autophagy in non-small cell lung cancer A549 cells based on mTOR/STAT3 signaling axis. Methods A549 cells were divided into blank group and 30，60，90， 120 and 150 μg·mL-1 of morin groups. After 24， 48 and 72 hours of culture， the cell proliferation activity was detected by CCK-8 method，and the cell inhibition rate was calculated. A549 cells were divided into blank group and 30， 90， 150 μg·mL-1 morin groups. After 14 days of culture， the cell proliferation was detected by colony formation assay. After 24 hours of culture， the cell proliferation ability was detected by BeyoClickTM EdU-488. Apoptosis was detected by flow cytometry；acridine orange staining was used to detect cell autophagy；the formation of autophagosomes was observed by transmission electron microscopy. Western Blot was used to detect the expression levels of apoptosis， autophagy and mTOR/STAT3 signaling axis-related proteins in cells. A549 cells were divided into blank group，blank group + chloroquine（10 μg·mL-1） group，morin（30，150 μg·mL-1） group，morin（30， 150 μg · mL-1）+ chloroquine （10 μg·mL-1） group. After 48 hours of intervention，the cell activity was detected by CCK-8 method，and the cell survival rate was calculated. Results Compared with the blank group，the inhibition rate of A549 cells in 60， 90， 120， 150 μg · mL-1 of morin group was significantly increased after 24 hours of intervention （P＜0.05，P＜0.001） . The inhibition rates of A549 cells in 30，60，90，120 and 150 μg·mL-1 of morin groups were significantly increased after 48 and 72 hours of intervention （P＜0.001） . The number of A549 cell colonies and the number of green fluorescent proliferation positive cells in the 30，90，150 μg·mL-1 of morin groups were significantly decreased （P＜0.01，P＜0.001），the apoptosis rate was significantly increased （P＜0.01，P＜ 0.001）， and the protein expression level of cleaved-PARP was significantly increased （P＜0.001） . The protein expression levels of p-P38/P38 MAPK in A549 cells of 90 and 150 μg·mL-1 of morin groups were significantly increased （P＜0.01，P＜0.001） . Different degrees of orange fluorescence appeared in A549 cells of 30，90 and 150 μg·mL-1 of morin groups，and the orange fluorescence of 90 and 150 μg·mL-1 of morin groups was significant. Autophagosomes and autolysosomes appeared in the cytoplasm of A549 cells in 150 μg · mL-1 of morin group， respectively. The protein expression of LC3-Ⅱ in A549 cells of 150 μg·mL-1 of morin group was significantly upregulated（ P＜0.05） . The protein expression of Atg16L1-Ⅱ in A549 cells of 90，150 μg·mL-1 of morin group was significantly up-regulated （P ＜ 0.001）， and the protein expressions of p-mTOR/mTOR and p-STAT3/STAT3 were significantly down-regulated （P＜0.001） . Compared with the morin （150 μg·mL-1） group，the survival rate of A549 cells in the morin （150 μg·mL-1） +chloroquine （10 μg·mL-1） group was significantly increased （P＜ 0.05） . Conclusion Morin can promote the apoptosis of A549 cells and induce autophagy in A549 cells，and the mechanism may be related to mTOR/STAT3 axis.

R285.5

国家重点研发计划项目（2019YFC1711400）。