黄柏碱调节AMPK/FOXO3a信号通路对非小细胞肺癌细胞增殖、侵袭和上皮间质转化影响的研究OA
Reserch of the effects of phellodendrine adjusting the AMPK/FOXO3a signaling pathway on the proliferation,invasion and epithelial-mesenchymal transition of non-small cell lung cancer cells
目的 探究黄柏碱调节AMP活化蛋白激酶(AMPK)/叉头盒蛋白O3(FOXO3a)信号通路对非小细胞肺癌细胞增殖、侵袭及上皮间质转化的影响.方法 将人非小细胞肺癌细胞NCI-H1650分为对照组、低剂量实验组、中剂量实验组、高剂量实验组和抑制剂组.对照组细胞用含等体积磷酸盐缓冲液(PBS)的培养基培养,低剂量实验组、中剂量实验组、高剂量实验组分别采用含有10、20、40 μmol·L-1黄柏碱的培养基培养,抑制剂组采用含有40 μmol·L-1黄柏碱和10 μmol·L-1 Dorsomorphin培养.用5-乙炔基-2′-脱氧尿苷(EdU)检测细胞的增殖能力,用流式细胞术检测细胞的凋亡能力,用Transwell检测细菌的迁移和侵袭能力,用蛋白质印迹法(Western blot)检测上皮间质转化蛋白E-钙粘蛋白(E-cadherin)、N-钙粘蛋白(N-cadherin)、波形蛋白(Vimentin)及AMPK/FOXO3a通路蛋白相对表达水平.结果 对照组、低剂量实验组、中剂量实验组、高剂量实验组和抑制剂组的EdU阳性率分别为(37.95±4.57)％、(30.14±3.73)％、(22.86±2.95)％、(15.34±1.97)％和(33.48±4.02)％,细胞凋亡率分别为(2.84±0.39)％、(9.46±1.25)％、(16.32±1.97)％、(23.01±2.74)％和(5.98±0.86)％,侵袭细胞数分别为(73.17±8.89)、(58.00±6.87)、(43.67±5.54)、(29.83±3.97)和(67.17±7.73)个,E-cadherin蛋白相对表达水平分别为1.05±0.12、1.35±0.16、1.64±0.19、1.92±0.23 和 1.14±0.14,N-cadherin 蛋白相对表达水平分别为 1.03±0.11、0.79±0.09、0.56±0.07、0.34±0.05 和 0.95±0.12,Vimentin蛋白相对表达水平分别为 1.06±0.12、0.82±0.10、0.58±0.08、0.35±0.06 和0.97±0.13,磷酸化(p)-AMPK/AMPK 水平分别为 0.38±0.05、0.55±0.07、0.71±0.09、0.88±0.10 和 0.41±0.05,p-FOXO3a/FOXO3a 分别为0.33±0.04、0.51±0.06、0.68±0.08、0.85±0.09 和 0.35±0.04.低剂量实验组、中剂量实验组、高剂量实验组、抑制剂组NCI-H1650细胞EdU阳性率、迁移和侵袭细胞数、凋亡率、E-cadherin、N-cadherin和Vimentin蛋白表达以及p-AMPK和p-FOXO3a水平比较,在统计学上差异有统计学意义(P＜0.05)o结论 黄柏碱通过激活AMPK/FOXO3a信号通路抑制非小细胞肺癌细胞增殖侵袭和上皮间质转化,并促进凋亡.
Objective To explore the effects of phellodendrine adjusting the AMP-activated protein kinase(AMPK)/forkhead box protein O3(FOXO3a)signaling pathway on the proliferation,invasion and epithelial-mesenchymal transition of non-small cell lung cancer cells.Methods Human non-small cell lung cancer cell line NCI-H1650 was divided into control group,low-dose experimental group,medium dose experimental group,high-dose experimental group,and an inhibitor group.The control group was cultured in medium containing an equal volume of phosphate-buffered saline(PBS).The low-dose experimental group,the medium dose experimental group,and the high-dose experimental group were cultured in media containing 10,20,and 40 μ mol·L-1 berberine,respectively.The inhibitor group was cultured in media containing 40 μ mol·L-1 berberine and 10 μ mol·L-1 Dorsomorphin.Use 5-Ethynyl-2′-deoxyuridine(EdU)to detect cell proliferation ability,use flow cytometry to detect cell apoptosis ability,use Transwell to detect cell migration and invasion ability,use Western blot to detect the relative expression levels of epithelial mesenchymal transition proteins E-cadherin,N-cadherin,vimentin,and AMPK/FOXO3a pathway proteins.Results The EdU positive rates of the control group,low-dose experimental group,medium dose experimental group,high-dose experimental group,and inhibitor group were(37.95±4.57)％,(30.14±3.73)％,(22.86±2.95)％,(15.34±1.97)％and(33.48±4.02)％,respectively;the cell apoptosis rates were(2.84±0.39)％,(9.46±1.25)％,(16.32±1.97)％,(23.01±2.74)％and(5.98±0.86)％,respectively;the number of invasive cells were(73.17±8.89),(58.00±6.87),(43.67±5.54),(29.83±3.97)and(67.17±7.73),respectively;E-cadherin protein expression were 1.05±0.12,1.35±0.16,1.64±0.19,1.92±0.23 and 1.14±0.14,respectively;N-cadherin protein expression were 1.03±0.11,0.79±0.09,0.56±0.07,0.34±0.05 and 0.95±0.12,respectively;Vimentin protein expression were 1.06±0.12,0.82±0.10,0.58±0.08,0.35±0.06 and 0.97±0.13,respectively;the phosphorylated(p)-AMPK/AMPK levels were 0.38±0.05,0.55±0.07,0.71±0.09,0.88±0.10 and 0.41±0.05,respectively;the p-FOXO3a/FOXO3a levels were 0.33±0.04,0.51±0.06,0.68±0.08,0.85±0.09 and 0.35±0.04,respectively.There were statistically significant differences in the EdU positivity rate,migration and invasion cell numbers,apoptosis rate,E-cadherin,N-cadherin and Vimentin protein expression,as well as p-AMPK and p-FOXO3a levels of NCI-H1650 cells between the low-dose experimental group,medium dose experimental group,high-dose experimental group,and inhibitor group.Conclusion Phellodendrine inhibits the proliferation,invasion and epithelial-mesenchymal transition of non-small cell lung cancer cells,and promotes apoptosis by activating the AMPK/FOXO3a signaling pathway.
任维敏;张宝;张国臣;乔鹏涛
济南市中西医结合医院,胸外两腺外科,山东济南 271100济南市中西医结合医院,胸外两腺外科,山东济南 271100济南市中西医结合医院,外科,山东济南 271100济南市中西医结合医院,胸外两腺外科,山东济南 271100
医药卫生
黄柏碱AMP活化蛋白激酶/叉头盒蛋白O3非小细胞肺癌增殖侵袭上皮间质转化
phellodendrineAMP-activated protein kinase/forkhead box protein O3non-small cell lung cancerproliferationinvasionepithelial-mesenchymal transition
《中国临床药理学杂志》 2026 (16)
2330-2336,7
济南市卫生健康委员会科技发展计划基金资助项目(2024302016)
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