【摘要】 目的:探讨山奈酚对鼻咽癌细胞增殖、凋亡和上皮间质转化(EMT)的影响，并探究其作用机制。方法:将CNE-2细胞分为对照组、山奈酚低浓度组、山奈酚中浓度组、山奈酚高浓度组、山奈酚高浓度+SC-79(AKT激活剂)组。CCK-8法和克隆实验测定细胞增殖，流式细胞术检测细胞凋亡，Western blot检测各蛋白B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、天冬氨酸蛋白水解酶-3(Caspase-3)、波形蛋白(Vimentin)、上皮钙黏蛋白(E-cadherin)、神经钙黏蛋白(N-cadherin)、蛋白激酶B(AKT)、磷酸化Akt(p-AKT)、糖原合成酶激酶3β(GSK-3β)、磷酸化GSK-3β(p-GSK-3β)、Snail的表达水平。构建鼻咽癌裸鼠模型，分为裸鼠NC组、山奈酚组、山奈酚+SC-79组，测量肿瘤质量与体积，免疫组化法检测移植瘤组织p-AKT、p-GSK-3β、Snail蛋白表达。结果:山奈酚对人鼻咽癌上皮细胞活力无显著影响(P>0.05);鼻咽癌细胞活力随着山奈酚浓度的升高而逐渐降低(P<0.05),选择CNE-2作为后续实验细胞，选择25、50、100μmol/L山奈酚作为后续实验浓度。与对照组相比，山奈酚低、中、高组细胞活力、Bcl-2、N-cadherin、Vimentin、 p-AKT/AKT、p-GSK-3β/GSK-3β、Snail水平显著下降，凋亡率、Bax、Caspase-3、E-cadherin上升(P<0.05);与山奈酚高浓度组相比，山奈酚高浓度+SC-79组细胞活力、Bcl-2、N-cadherin、Vimentin、p-AKT/AKT、p-GSK-3β/GSK-3β、Snail水平显著上升，凋亡率、Bax、Caspase-3、E-cadherin下降(P<0.05)。山奈酚能抑制移植瘤质量和体积，降低p-AKT、p-GSK-3β、Snail蛋白表达(P<0.05);SC-79逆转山奈酚对移植瘤的抑制作用，促进AKT/GSK-3β/Snail通路蛋白表达(P<0.05)。结论:山奈酚能抑制鼻咽癌细胞增殖和EMT,促进细胞凋亡，其作用机制可能与抑制AKT/GSK-3β/Snail信号通路有关。更多还原

【Abstract】 Objective:To investigate the impacts of kaempferol on proliferation,apoptosis,and epithelial-mesenchymal transition(EMT) of nasopharyngeal carcinoma cells,and to explore its mechanism of action.Methods:CNE-2 cells were grouped into a control group,a low concentration kaempferol group,a medium concentration kaempferol group,a high concentration kaempferol group,and a high concentration kaempferol+SC-79(AKT activator) group.CCK-8 method and cloning experiment were applied to measure cell proliferation,flow cytometry was applied to detect cell apoptosis,Weatern blot detected the expression level of B cell lymphoblastoma 2(Bcl-2),Bcl-2 associated X protein(Bax),aspartate proteolytic enzyme 3(Caspase-3),Vimentin,epithelial cadherin(E-cadherin),neural cadherin(N-cadberin),protein kinase B(AKT),phosphorylated Akt(p-AKT),glycogen synthase kinase-3β(GSK-3β),phosphorylated GSK-3β(p-GSK-3β),and Snail.A nude mouse model of nasopharyngeal carcinoma was constructed and grouped into NC group,kaempferol group,and kaempferol+SC-79 group.The tumor mass and volume were measured,and the expression of p-AKT,p-GSK-3β,and Snail protein in the transplanted tumor tissue was detected using immunohistochemistry.Results:Kaempferol had no significant effect on the viability of human nasopharyngeal carcinoma epithelial cells(P> 0.05).The vitality of nasopharyngeal carcinoma cells gradually decreased with the increase of kaempferol concentration(P <0.05).CNE-2 was selected as the subsequent experimental cell,and 25,50 and 100 μmol/L kaempferol were selected as the subsequent experimental concentrations.Compared with the control group,the cell viability,Bcl-2,N-cadherin,Vimentin,p-AKT/AKT,p-GSK-3β/GSK-3β,and Snail levels in the low,medium,and high kaempferol groups obviously decreased,and the apoptosis rate,the level of Bax,Caspase-3,and E-cadherin increased(P <0.05).Compared with the high concentration kaempferol group,the cell viability,Bcl-2,N-cadherin,Vimentin,p-AKT/AKT,p-GSK-3β/GSK-3β,and Snail levels in the high concentration kaempferol+SC-79 group obviously increased,and the apoptosis rate,the level of Bax,Caspase-3,and E-cadherin decreased(P <0.05).Kaempferol was able to inhibit the mass and volume of transplanted tumors,reduce the expression of p-AKT,p-GSK-3β,and Snail proteins(P <0.05).SC-79 reversed the inhibitory effect of kaempferol on transplanted tumors and promoted the expression of AKT/GSK-3β/Snail pathway proteins(P <0.05).Conclusion:Kaempferol can inhibit the proliferation and EMT of nasopharyngeal carcinoma cells,promote cell apoptosis,and its mechanism of action may be related to the inhibition of AKT/GSK-3β/Snail signaling pathway.更多还原