敲减抗增殖蛋白2促进非小细胞肺癌细胞系A549凋亡

doi:10.16352/j.issn.1001-6325.2023.10.1522

基础医学与临床 ›› 2023, Vol. 43 ›› Issue (10): 1522-1529.doi: 10.16352/j.issn.1001-6325.2023.10.1522

敲减抗增殖蛋白2促进非小细胞肺癌细胞系A549凋亡

张婧¹, 杨自更², 韦红梅¹, 宁海虹³, 薛茜茜¹, 金玲⁴, 吴宾^4*

新疆军区总医院 1.营养科;
2.核医学科;
3.卫勤训练中心;
4.老年医学科,新疆乌鲁木齐 830000

收稿日期:2023-03-01 修回日期:2023-06-01 出版日期:2023-10-05 发布日期:2023-10-05
通讯作者: *wubin0917@163.com
基金资助:
新疆维吾尔族自治区自然科学基金(2022D01C644);新疆军区总医院喀喇昆仑基金(2022JC002)

Knockdown of prohibitin 2 promotes apoptosis in non-small cell lung cancer cell line A549

ZHANG Jing¹, YANG Zigeng², WEI Hongmei¹, NING Haihong³, XUE Xixi¹, JIN Ling⁴, WU Bin^4*

1. Department of Nutrition;
2. Department of Nuclear Medicine;
3. Training Center of Health Service;
4. Department of Geriatrics,General Hospital of Xinjiang Military Command, Urumqi 830000, China

Received:2023-03-01 Revised:2023-06-01 Online:2023-10-05 Published:2023-10-05
Contact: *wubin0917@163.com

摘要/Abstract

摘要： 目的探讨抑制抗增殖蛋白2(PHB2)表达对非小细胞肺癌细胞系A549凋亡的影响及机制。方法慢病毒感染非小细胞肺癌细胞系A549,建立敲减PHB2的稳转细胞株,用动力相关蛋白1(DRP1)抑制剂Mdivi-1处理敲减PHB2的A549细胞。Western blot检测PHB2、p-DRP1(Ser616)和DRP1蛋白表达水平;TUNEL染色和annexin V-FITC/PI双染检测细胞凋亡;MitoTracker染色评估线粒体分裂情况;用相应试剂盒检测线粒体含量、柠檬酸合酶活性和细胞色素c含量。结果与shCtrl组相比,shPHB2组A549细胞凋亡率增加(P＜0.05),DRP1介导的线粒体分裂增加(P＜0.05),ATP含量减少(P＜0.05),柠檬酸合酶活性降低(P＜0.05),线粒体内细胞色素c减少(P＜0.05)。与shPHB2组相比,shPHB2+Mdivi-1组A549细胞凋亡率减少(P＜0.05),DRP1介导的线粒体分裂减少(P＜0.05),ATP含量增加(P＜0.05),柠檬酸合酶活性增加(P＜0.05),线粒体内细胞色素c增加(P＜0.05)。结论抑制PHB2促进A549细胞凋亡,其机制可能与促进DRP1介导的线粒体分裂有关。

关键词: 抗增殖蛋白2, 线粒体分裂, 非小细胞肺癌, 动力相关蛋白1, 凋亡

Abstract: Objective To explore mechanism of prohibitin 2(PHB2) inhibition on the apoptosis of non-small cell lung cancer cell line A549. Methods For stable knockdown of PHB2, A549 strain was infected with lentiviruses. A549 cells with PHB2 depletion were incubated with Mdivi-1(DRP1 inhibitor). Apoptotic index of A549 cells was evaluated by TUNEL and annexin V-FITC/PI. Mitochondrial morphology was visualized by MitoTracker. ATP content, citrate synthase activity and the level of cytochrome c in cytosolic and mitochondrial fractions were analyzed by commercially available kits. Western blot was used to measure the levels of PHB2, DRP1 and p-DRP1 protein expression. Results Compared with the shCtrl group, the apoptosis of A549 cells increased significantly (P＜0.05) with up-regulated DRP1-dependent mitochondrial fission (P＜0.05), down-regulated ATP content (P＜0.05) and citrate synthase activity (P＜0.05). The level of cytochrome C was decreased in mitochondrial fractions (P＜0.05). Compared with the shPHB2 group, the apoptosis of A549 cells decreased significantly (P＜0.05) with alleviated DRP1-dependent mitochondrial fission (P＜0.05), increase of ATP content (P＜0.05) and citrate synthase activity(P＜0.05). The level of cytochrome C was increased in mitochondrial fractions (P＜0.05) in the shPHB2+Mdivi-1 group. Conclusions PHB2 inhibition can significantly enhance the apoptosis rate of A549 cells, potentially through promoting DRP1-dependent mitochondrial fission.

Key words: prohibitin 2(PHB2), mitochondrial fission, non-small cell lung cancer, dynamin-related protein 1(DRP1), apoptosis

中图分类号:

R734.2

张婧, 杨自更, 韦红梅, 宁海虹, 薛茜茜, 金玲, 吴宾. 敲减抗增殖蛋白2促进非小细胞肺癌细胞系A549凋亡[J]. 基础医学与临床, 2023, 43(10): 1522-1529.

ZHANG Jing, YANG Zigeng, WEI Hongmei, NING Haihong, XUE Xixi, JIN Ling, WU Bin. Knockdown of prohibitin 2 promotes apoptosis in non-small cell lung cancer cell line A549[J]. Basic & Clinical Medicine, 2023, 43(10): 1522-1529.

参考文献 15

[1]	Siegel RL, Miller KD, Fuchs HE, et al. Cancer statistics, 2021[J]. CA Cancer J Clin, 2021,71:7-33.
[2]	Sung H, Ferlay J, Siegel RL, et al. Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries[J]. CA Cancer J Clin, 2021,71:209-249.
[3]	薛剑超, 王亚东, 李博文, 等. DNA甲基化在肺癌发生及预后中作用的研究进展[J]. 基础医学与临床, 2023,43:514-518.
[4]	任孟先, 梁伟, 刘艳. 塞来昔布联合IMRT对老年中晚期非小细胞肺癌患者免疫功能和血清肿瘤标志物的影响[J]. 基础医学与临床, 2021,41:890-894.
[5]	Signorile A, Sgaramella G, Bellomo F, et al. Prohibitins: a critical role in mitochondrial functions and implication in diseases[J]. Cells, 2019,8:278-286.
[6]	Wu B, Chang N, Xi H, et al. PHB2 promotes tumorigenesis via RACK1 in non-small cell lung cancer[J]. Theranostics, 2021,11:3150-3166.
[7]	Kasashima K, Ohta E, Kagawa Y, et al. Mitochondrial functions and estrogen receptor-dependent nuclear translocation of pleiotropic human prohibitin 2[J]. J Biol Chem, 2006,281:36401-36410.
[8]	Cai XW, Yu WW, Yu W, et al. Tissue-based quantita-tive proteomics to screen and identify the potential biomarkers for early recurrence/metastasis of esophageal squamous cell carcinoma[J]. Cancer Med, 2018,7:2504-2517.
[9]	Cheng J, Gao F, Chen X, et al. Prohibitin-2 promotes hepatocellular carcinoma malignancy progression in hypoxia based on a label-free quantitative proteomics strategy[J]. Mol Carcinog, 2014,53:820-832.
[10]	Cao Y, Liang H, Zhang F, et al. Prohibitin overexpres-sion predicts poor prognosis and promotes cell proliferation and invasion through ERK pathway activation in gallbl-adder cancer[J]. J Exp Clin Cancer Res, 2016,35:68-78.
[11]	Yapa N, Lisnyak V, Reljic B, et al. Mitochondrial dynamics in health and disease[J]. FEBS Lett, 2021,595:1184-1204.
[12]	Srinivasan S, Guha M, Kashina A, et al. Mitochondrial dysfunction and mitochondrial dynamics-the cancer connection[J]. Biochim Biophys Acta Bioenerg, 2017,1858:602-614.
[13]	Willems PH, Rossignol R, Dieteren CE, et al. Redox homeostasis and mitochondrial dynamics[J]. Cell Metab, 2015,22:207-218.
[14]	Serasinghe MN, Chipuk JE. Mitochondrial fission in human diseases[J]. Handb Exp Pharmacol, 2017,240:159-188.
[15]	Wu B, Xiong J, Zhou Y, et al. Luteolin enhances TRAIL sensitivity in non-small cell lung cancer cells through increasing DR5 expression and Drp1-mediated mitochondrial fission[J]. Arch Biochem Biophys, 2020,692:108539-108543.

敲减抗增殖蛋白2促进非小细胞肺癌细胞系A549凋亡

Knockdown of prohibitin 2 promotes apoptosis in non-small cell lung cancer cell line A549

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献 15

相关文章 15

编辑推荐

Metrics

本文评价

[1]	罗梦琳, 郑峰, 季欣瑶, 祁彩虹, 肖雨珩, 牛长春. 流体剪切力通过激活Piezo 1蛋白促进肾小球内皮细胞凋亡[J]. 基础医学与临床, 2024, 44(9): 1236-1242.
[2]	王婷婷, 魏欢, 杨永丽, 周贤, 胡阳. 三七皂苷Rg1对阿尔茨海默症大鼠的神经系统的保护作用[J]. 基础医学与临床, 2024, 44(8): 1094-1100.
[3]	高超, 张润菡, 王伟, 赵曼婷, 焦艳, 李喆. 青蒿琥酯调节cGAS-STING信号通路对抑郁症模型小鼠神经炎性反应的影响[J]. 基础医学与临床, 2024, 44(8): 1126-1132.
[4]	郁冬玲, 莫少娥, 傅文, 陈实, 谢酬勤, 蓝雨雁. 抑制sigma-1受体可减少神经病理性疼痛大鼠DRG细胞凋亡[J]. 基础医学与临床, 2024, 44(8): 1101-1106.
[5]	郑红, 陈晓霞, 韩李周, 陈苗, 皇甫娟. 降低lncRNA-RMRP表达抑制人肺癌细胞系A549的增殖和侵袭[J]. 基础医学与临床, 2024, 44(7): 974-978.
[6]	高寅洁, 童安莉. 醛固酮产生腺瘤中细胞死亡和增殖机制[J]. 基础医学与临床, 2024, 44(6): 758-762.
[7]	李晋平, 刘汉卿, 杨全会. 大鼠严重腹腔感染后脾脏细胞周期的变化[J]. 基础医学与临床, 2024, 44(6): 828-832.
[8]	龚金涛, 厉建伟, 李玉恒, 李堑, 赵春华. 骨髓间充质干细胞缓解模拟微重力对小鼠大脑皮质的不利影响[J]. 基础医学与临床, 2024, 44(6): 772-778.
[9]	梁香存, 魏小雨, 梁健, 王庆, 耿广. 安罗替尼通过调控miR-16-5p/PD-1轴抑制人非小细胞肺癌细胞系增殖并促进凋亡[J]. 基础医学与临床, 2024, 44(4): 503-512.
[10]	杨春生, 王添兴, 张铁成, 武恒敏, 王宝兰. 软骨细胞中生物钟基因Bmal1对细胞周期相关基因表达的影响[J]. 基础医学与临床, 2024, 44(4): 496-502.
[11]	杨铠菲, 朱静格, 张洋洋, 赵俊果, 高雨悦, 胡焕焕, 姬国杰. 索拉菲尼诱导的细胞凋亡和自噬对HeLa细胞耐药性的影响[J]. 基础医学与临床, 2024, 44(4): 467-473.
[12]	李娜, 李涛, 杨冬梨, 姚媛. 和厚朴酚抑制人脂肪间充质干细胞增殖[J]. 基础医学与临床, 2024, 44(4): 483-488.
[13]	张婧, 杨自更, 蔡文琴, 曹维维, 韦红梅, 薛茜茜, 吴宾. 抑制抗增殖蛋白2增强非小细胞肺癌细胞系A549对厄洛替尼敏感性[J]. 基础医学与临床, 2024, 44(3): 325-332.
[14]	安琪, 齐光照, 韩超. 桃叶珊瑚苷抑制人肝癌细胞系HepG2增殖[J]. 基础医学与临床, 2024, 44(3): 333-338.
[15]	李娟, 李晓峰, 徐生志, 唐凯. 华蟾素胶囊联合多西紫杉醇+顺铂化疗用于非小细胞肺癌患者的疗效[J]. 基础医学与临床, 2024, 44(2): 247-251.