莪术醇通过调控PI3K/AKT通路促进人肝癌HepG2细胞凋亡
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摘要
目的探讨莪术醇促进人肝癌HepG2细胞凋亡的作用是否与PI3K/AKT通路有关。方法采用MTT法检测不同浓度(0、10、25、50、100μmol·L~(-1))莪术醇干预HepG2细胞24、48 h的增殖抑制率,Hoechst 33258染色法观察细胞核凋亡形态,Western blot法检测不同浓度莪术醇及PI3K/AKT通路阻断剂LY294002对HepG2细胞p-PI3K、p-AKT、Caspase-3蛋白表达的影响。结果莪术醇对HepG2细胞的增殖有明显抑制作用,呈浓度依赖性,但干预48 h对比24 h差异无统计学意义;Hoechst 33258染色发现,莪术醇干预后,同一视野下细胞数目明显减少,细胞透亮,部分细胞核碎裂,核固缩,呈现典型的凋亡形态学特征;Western blot结果显示,莪术醇能显著降低细胞p-PI3K、p-AKT蛋白的表达,上调凋亡执行蛋白Caspase-3的表达,与LY294002联用后表现出良好的协同作用,对各蛋白表达的调控较单独用药更加明显。结论莪术醇能通过抑制PI3K/AKT通路诱导人肝癌HepG2细胞凋亡。
Objective To determine whether curcumol promotes the apoptosis of human hepatoma HepG2 cells and whether it is related to PI3 K/AKT pathway. Methods MTT assay was used to detect the proliferation inhibition rate of HepG2 cells treated with curcumol at different concentrations(0, 10, 25, 50, and 100 μmol·L~(-1)) for 24 h and 48 h. Nuclear apoptotic morphology was observed by Hoechst 33258 staining. Western blot was used to detect p-PI3 K, p-AKT, and Caspase-3 protein expression in different concentrations of curcumol and PI3 K/AKT pathway blocker LY294002. Results Curcumol significantly inhibited the proliferation of HepG2 cells in a concentration-dependent manner, but with no significant difference between the interventions at 48 h and 24 h. Hoechst 33258 staining showed that after the curcumol intervention, the number of cells in the same field was significantly reduced, the cells were translucent, part of the nuclei were fragmented, and nuclear condensation was observed, indicating typical apoptotic morphological features. Western blot showed that curcumol significantly reduced the expression of p-PI3 K and p-AKT proteins, up-regulated the expression of apoptosis executive protein Caspase-3. After combination with LY294002, curcumol showed good synergistic effect, and the regulation of each protein expression was more obvious than the use of drugs alone. Conclusion Curcumol induces the apoptosis of human hepatocellular HepG2 cells through the inhibition of PI3 K/AKT pathway.
Objective To determine whether curcumol promotes the apoptosis of human hepatoma HepG2 cells and whether it is related to PI3 K/AKT pathway. Methods MTT assay was used to detect the proliferation inhibition rate of HepG2 cells treated with curcumol at different concentrations(0, 10, 25, 50, and 100 μmol·L~(-1)) for 24 h and 48 h. Nuclear apoptotic morphology was observed by Hoechst 33258 staining. Western blot was used to detect p-PI3 K, p-AKT, and Caspase-3 protein expression in different concentrations of curcumol and PI3 K/AKT pathway blocker LY294002. Results Curcumol significantly inhibited the proliferation of HepG2 cells in a concentration-dependent manner, but with no significant difference between the interventions at 48 h and 24 h. Hoechst 33258 staining showed that after the curcumol intervention, the number of cells in the same field was significantly reduced, the cells were translucent, part of the nuclei were fragmented, and nuclear condensation was observed, indicating typical apoptotic morphological features. Western blot showed that curcumol significantly reduced the expression of p-PI3 K and p-AKT proteins, up-regulated the expression of apoptosis executive protein Caspase-3. After combination with LY294002, curcumol showed good synergistic effect, and the regulation of each protein expression was more obvious than the use of drugs alone. Conclusion Curcumol induces the apoptosis of human hepatocellular HepG2 cells through the inhibition of PI3 K/AKT pathway.
引文
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[2]Zhang C,Wang LM. Inhibition of autophagy attenuated curcumol-induced apoptosis in MG-63 human osteosarcoma cells via Janus kinase signaling pathway[J]. Oncol Lett,2017,14(6):6387-6394.
[3]NingL,MaH,JiangZ,etal.Curcumolsuppresses breast cancer cell metastasis by inhibiting MMP-9 via JNK1/2 and Akt-dependent NF-κB signaling pathways[J].Integr Cancer Ther,2016,15(2):216-225.
[4]Barraco D,Mudireddy M,Shah S,et al. Liver function test abnormalities and their clinical relevance in primary myelofibrosis[J]. Blood Cancer J,2017,7(4):557-562.
[5]杨全会,许荣焜.细胞凋亡与肿瘤[J].生理科学进展,2006,37(4):373-377.
[6]韩欣月,王梓,李伟.人参茎叶总皂苷对顺铂致小鼠肾损伤的保护作用及机制[J].中国药理学与毒理学杂志,2017,31(2):151-158.
[7]Bommareddy A,Crisamore K,Fillman S,et al. Survivin down-regulation byα-santalol is not mediated through PI3K-AKT pathway in human breast cancer cells[J]. Anticancer Res,2015,35(10):5353-5357.
[8]Astle F. Diabetes and depression:a review of the literature[J]. Nurs Clin North Am,2007,42(1):67-78.
[9]韦立群,李清,甘嘉亮,等.迷迭香酸衍生物RAD9通过PI3K/AKT和p38MAPK信号通路诱导胃癌MGC-803细胞凋亡[J].中国药理学通报,2018,34(2):256-260.
[10]Yang M,Wang H,Zhou M. The natural compound sulforaphene,as a novel anticancer reagent,targeting PI3KAKT signaling pathway in lung cancer[J]. Oncotarget,2016,47(7):76656-76666.
[11]Roy M,Tomar D,Singh K,et al. TRIM8 regulated autophagy modulates the level of cleaved Caspase-3 subunit to inhibit genotoxic stress induced cell death[J]. Cell Signal,2018,17(4):1-12.
[12]Malick M,Gilbert K,Daniel J,et al. Vagotomy prevents the effect of probiotics on caspase activity in a model of postmyocardial infarction depression[J]. Neurogastroenterol Motil,2015,27(5):663-671.
[13]张博.姜黄素、莪术醇对人肝癌细胞SMMC-7721 PI3K/AKT信号通路的影响[D].南京:南京中医药大学,2013.