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郭利培,刘洁,张文青,史红健,范婧莹,王贤文,何迎春.吴茱萸碱调控PI3K/AKT信号通路抑制鼻咽癌细胞增殖和诱导凋亡[J].湖南中医药大学学报,2023,43(4):612-618[点击复制] |
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| 吴茱萸碱调控PI3K/AKT信号通路抑制鼻咽癌细胞增殖和诱导凋亡 |
| 郭利培,刘洁,张文青,史红健,范婧莹,王贤文,何迎春 |
| (湖南中医药大学, 湖南 长沙 410208;湖南中医药大学, 湖南 长沙 410208;中医药防治眼耳鼻咽喉疾病湖南省重点实验室, 湖南 长沙 410208;湖南省中医药防治眼耳鼻咽喉疾病与视功能保护工程技术研究中心, 湖南 长沙 410208;中医药防治眼耳鼻咽喉疾病湖南省重点实验室, 湖南 长沙 410208;湖南省中医药防治眼耳鼻咽喉疾病与视功能保护工程技术研究中心, 湖南 长沙 410208;湖南中医药大学第一附属医院, 湖南 长沙 410007) |
| 摘要: |
| 目的 研究吴茱萸碱对鼻咽癌5-8F细胞增殖、凋亡及PI3K/AKT信号通路的影响。方法 （1）将5-8F细胞分为溶剂对照组、不同浓度吴茱萸碱组（0.5、1.0、2.0、4.0 μmol·L-1）、顺铂组（cisplatin，4.0 μg·mL-1）；（2）将5-8F细胞设为5组：溶剂对照组、SC79 2.5 μmol·L-1组、SC79+吴茱萸碱 2.0 μmol·L-1组 、吴茱萸碱 2.0 μmol·L-1组、LY294002 50 μmol·L-1组。采用实时无标记细胞功能分析仪（real time cellular analysis technology, RTCA）监测细胞增殖的情况；Annexin V-FITC/PI双荧光染色法检测细胞凋亡率，Hoechest 33342染色法观察细胞凋亡形态；Western blot法检测磷酸肌醇3-激酶蛋白(phosphatidylinositol 3-kiases, PI3K)、磷酸化蛋白质激酶蛋白B(phosphorylated protein kinase B, p-AKT)、增殖细胞核抗原蛋白（proliferating cell nuclear antigen, PCNA）、X连锁凋亡抑制蛋白（X-linked inhibitor of apoptosis, XIAP）、存活蛋白（Survivin）的表达水平。结果 与溶剂对照组相比，不同浓度吴茱萸碱组（0.5、1.0、2.0、4.0 μmol·L-1）显著抑制5-8F细胞增殖（P＜0.05或P<0.01）；吴茱萸碱组（1.0、2.0 μmol·L-1）的细胞核荧光染色增强，凋亡率明显升高（P<0.01）；PI3K、p-AKT、PCNA、XIAP、Survivin蛋白表达水平明显下降（P＜0.05或P＜0.01）。与吴茱萸碱2.0 μmol·L-1组比较，SC79+吴茱萸碱2.0 μmol·L-1组的PI3K、p-AKT、PCNA、XIAP、Survivin蛋白表达水平升高（P＜0.05或P＜0.01），且吴茱萸碱抑制5-8F细胞增殖和诱导凋亡的效应降低（P＜0.05或P＜0.01）。结论 吴茱萸碱可能通过抑制PI3K/AKT信号通路活性，下调增殖及凋亡相关蛋白PCNA、XIAP、Survivin的表达水平，最终抑制鼻咽癌细胞增殖和诱导凋亡。 |
| 关键词: 鼻咽癌细胞 吴茱萸碱 增殖 凋亡 PI3K/AKT信号通路 |
| DOI：10.3969/j.issn.1674-070X.2023.04.006 |
| 投稿时间：2022-11-07 |
| 基金项目:国家自然科学基金项目（81973914，81874408）；湖南省教育厅项目（21B0358，21C0241）；湖南省中医药管理局项目（D2022105）；2021年度湖南中医药大学校级科研基金项目（2021XJJJ014，2021XJJJ008）；湖南省卫生健康委员会项目（D202307017740）。 |
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| Regulation of evodiamine on PI3K/AKT signaling pathway to inhibit proliferation and induce apoptosis of nasopharyngeal carcinoma cells |
| GUO Lipei,LIU Jie,ZHANG Wenqing,SHI Hongjian,FAN Jingying,WANG Xianwen,HE Yingchun |
| (Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;Hunan Provincial Key Laboratory for the Prevention and Treatment of Ophthalmology and Otolaryngology Diseases with Chinese Medicine, Changsha, Hunan 410208, China;Hunan Provincial Ophthalmology and Otolaryngology Diseases Prevention and Treatment with Chinese Medicine and Visual Function Protection Engineering and Technological Research Center, Changsha, Hunan 410208, China;Hunan Provincial Key Laboratory for the Prevention and Treatment of Ophthalmology and Otolaryngology Diseases with Chinese Medicine, Changsha, Hunan 410208, China;Hunan Provincial Ophthalmology and Otolaryngology Diseases Prevention and Treatment with Chinese Medicine and Visual Function Protection Engineering and Technological Research Center, Changsha, Hunan 410208, China;The First Hospital of Hunan University of Chinese Medicine, Changsha, Hunan 410007, China) |
| Abstract: |
| Objective To investigate the effects of evodiamine on proliferation and apoptosis of 5-8F cells in nasopharyngeal carcinoma cells and the role of PI3K/AKT signaling pathway.Methods The 5-8F cells were divided into solvent control group, evodiamine (0.5, 1.0, 2.0, 4.0 μmol·L-1) groups and cisplatin （4.0 μgmol·L-1） group; in addition, the 5-8F cells were divided into 5 groups: solvent control group, SC79 (2.5 μmol ·L-1) group, SC79+evodiamine (2.0 μmol·L-1) group, evodiamine (2.0 μmol·L-1) group, and LY294002 (50 μmol·L-1) group. The cell proliferation was monitored by real time cellular analysis technology (RTCA), the apoptosis rate was detected by annexin V-FITC/PI double fluorescence staining, and the apoptosis morphology was observed by Hoechest 33342 staining. The expression levels of phosphatidylinositol 3-kinase (PI3K), phosphorylated protein kinase B (p-AKT), proliferating cell nuclear antigen (PCNA), X-linked inhibitor of apoptosis (XIAP) and Survivin were detected by Western blot.Results Compared with the solvent control group, different concentrations of evodiamine (0.5, 1.0, 2.0, 4.0 μmol·L-1) significantly inhibited the proliferation of 5-8F cells (P<0.05 or P<0.01), and the nuclear fluorescence staining and apoptosis rate of cells in evodiamine group (1.0, 2.0 μmol·L-1) increased significantly (P<0.01). The protein expression of PI3K, p-AKT, PCNA, XIAP and Survivin decreased significantly (P<0.05 or P<0.01). Compared with evodiamine (2.0 μmol·L-1) group, the protein expression levels of PI3K, p-AKT, PCNA, XIAP and Survivin in SC79+evodiamine (2.0 μmol·L-1) group were elevated (P<0.05 or P<0.01), and the effects of evodiamine on inhibiting proliferation and inducing apoptosis of 5-8F cells decreased (P<0.05 or P<0.01).Conclusion Evodiamine may inhibit proliferation and induce apoptosis of nasopharyngeal carcinoma cells by inhibiting the activity of PI3K/AKT signaling pathway and down-regulating the expression of proliferation and apoptosis-related proteins PCNA, XIAP and Survivin. |
| Key words: nasopharyngeal carcinoma cell evodiamine proliferation apoptosis PI3K/AKT signaling pathway |
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