AMPK活性对HeLa细胞内质网功能稳态的影响

doi:10.3969/j.issn.1004-616x.2015.03.003

癌变·畸变·突变 ›› 2015, Vol. 27 ›› Issue (3): 172-176.doi: 10.3969/j.issn.1004-616x.2015.03.003

AMPK活性对HeLa细胞内质网功能稳态的影响

梁斌, 袁松, 刘艳敏, 宋旭红

汕头大学医学院细胞生物学与遗传学教研室, 广东汕头 515041

收稿日期:2015-03-12 修回日期:2015-04-14 出版日期:2015-05-30 发布日期:2015-05-30
作者简介:梁斌,E-mail:bliang@stu.edu.cn
基金资助:
国家自然科学基金(81470441);广东省自然科学基金(2014A030313473)

Effects of AMP-activated protein kinase activation on homeostasis of endoplasmic reticulum function in HeLa cells

LIANG Bin, YUAN Song, LIU Yanmin, SONG Xuhong

Department of Cell Biology and Genetics, Shantou University Medical College, Shantou 515041, Guangdong, China

Received:2015-03-12 Revised:2015-04-14 Online:2015-05-30 Published:2015-05-30

摘要/Abstract

摘要：

目的:探讨腺苷酸活化蛋白激酶(AMPK)对HeLa细胞内质网应激的影响。方法:试验分为对照组(1‰ DMSO)、内质网应激诱导组(2 μg/mL衣霉素或500 nmol/L MG132内质网应激诱导剂诱导)和内质网应激干预组[衣霉素或MG132+2 mmol/L AMPK激动剂5-氨基咪唑-4-甲酰胺-1-B-呋喃核糖苷(AICAR)干预组、衣霉素或MG132+2 mmol/L AMPK激动剂二甲双胍干预组以及衣霉素或MG132+0.5 mmol/L AMPK抑制剂Compound C干预组],以探究各处理组中AMPK活性状态对内质网应激的影响。收集对数生长期的HeLa细胞,分组诱导孵育后,用Western blot检测HeLa细胞AMPK/T-172磷酸化水平及内质网应激标志蛋白(p-elf2α、Grp78和XBP-1s)的表达,并采用细胞内钙集群检测分析(Indo-1 ratiometric Ca²⁺ analysis)检测各组HeLa细胞内Ca²⁺浓度的变化。结果:衣霉素和MG132可诱导内质网应激标志蛋白p-elf2α、Grp78和XBP-1s的表达。AMPK激动剂AICAR和二甲双胍干预可降低上述3种内质网应激标志蛋白的表达,并降低细胞胞浆内因内质网应激导致的Ca²⁺浓度升高,与对照组比较,差异均有统计学意义(P均<0.05)。AMPK抑制剂Compound C的干预对上述内质网应激标志蛋白的表达无明显影响(P均>0.05)。结论:AMPK激活能缓解HeLa细胞中的内质网应激,维持细胞内质网的功能稳态,可能对提高肿瘤细胞的应激耐受能力起到主要作用。

关键词: 腺苷酸活化蛋白激酶, 肿瘤细胞, 内质网, 应激

Abstract:

OBJECTIVE: To observe the effects of AMPK activation on intracellular endoplasmic reticulum stress (ER stress) in Hela cells. METHODS:Cultured HeLa cells were divided into groups:1‰ DMSO control, tunicamycin (2 μg/mL) or MG132 (500 nmol/L), tunicamycin or MG132+Compound C (0.5 mmol/L);tunicamycin or MG132+ AICAR(2 mmol/L) and tunicamycin or MG132+metformin (2 mmol/L). AMPK activation and markers of ER stress were determined by Western blot, and then intracellular Ca²⁺ concentration was measured in HeLa cells by Indo-1 ratiometric Ca²⁺ analysis. RESULTS:Our results showed that tunicamycin and MG132 could increase expression of ER stress. Activation of AMPK by AICAR and metformin could reduce the expression of above markers, and decrease the concentration of intracellular Ca²⁺. CONCLUSION:AMPK activation could relieve intracellular ER stress, maintain the homeostasis of intracellular Ca²⁺ concentration, and further, may contribute to cellular adaptation and resistance to stress stimuli.

Key words: AMPK, cancer cells, endoplasmic reticulum, stress

中图分类号:

R730.2

梁斌, 袁松, 刘艳敏, 宋旭红. AMPK活性对HeLa细胞内质网功能稳态的影响[J]. 癌变·畸变·突变, 2015, 27(3): 172-176.

LIANG Bin, YUAN Song, LIU Yanmin, SONG Xuhong. Effects of AMP-activated protein kinase activation on homeostasis of endoplasmic reticulum function in HeLa cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2015, 27(3): 172-176.

参考文献

[1] Hotamisligil GS. Endoplasmic reticulum stress and the inflammatory basis of metabolic disease[J]. Cell, 2010, 140 (6):900-917.
[2] Zhang X, Zhang G, Zhang H, et al. Hypothalamic IKKbeta/ NF-kappaB and ER stress link overnutrition to energy imbalance and obesity[J]. Cell, 2008, 135(1):61-73.
[3] Song X, Huang D, Liu Y, et al. AMP-activated protein kinase is required for cell survival and growth in HeLa-S3 cells in vivo[J]. IUBMB Life, 2014, 66(6):415-423.
[4] Kahn BB, Alquier T, Carling D, et al. AMP-activated protein kinase:ancient energy gauge provides clues to modern understanding of metabolism[J]. Cell Metab, 2005, 1(1):15-25.
[5] Carling D, Thornton C, Woods A, et al. AMP-activated protein kinase:new regulation, new roles?[J]. Biochem J, 2012, 445(1):11-27.
[6] Faubert B, Boily G, Izreig S, et al. AMPK is a negative regulator of the Warburg effect and suppresses tumor growth in vivo[J]. Cell Metab, 2013, 17(1):113-124.
[7] Liang B, Song X, Liu G, et al. Involvement of TR3/Nur77 translocation to the endoplasmic reticulum in ER stress-induced apoptosis[J]. Exp Cell Res, 2007, 313(13):2833-2844.
[8] Xin Q, Ji B, Cheng B, et al. Endoplasmic reticulum stress in cerebral ischemia[J]. Neurochem Int, 2014, 68:18-27.
[9] Hung JY, Hsu YL, Ni WC, et al. Oxidative and endoplasmic reticulum stress signaling are involved in dehydrocostuslactone-mediated apoptosis in human non-small cell lung cancer cells[J]. Lung Cancer, 2010, 68(3):355-365.
[10] Xu Y, Chiu JF, He QY, et al. Tubeimoside-1 exerts cytotoxicity in HeLa cells through mitochondrial dysfunction and endoplasmic reticulum stress pathways[J]. J Proteome Res, 2009, 8(3):1585-1593.
[11] Mujcic H, Rzymski T, Rouschop KM, et al. Hypoxic activation of the unfolded protein response (UPR) induces expression of the metastasis-associated gene LAMP3[J]. Radiother Oncol, 2009, 92(3):450-459.
[12] Jeon SM, Chandel NS, Hay N. AMPK regulates NADPH homeostasis to promote tumour cell survival during energy stress[J]. Nature, 2012, 485(7400):661-665.
[13] Demaurex N, Distelhorst C. Cell biology:Apoptosis——the calcium connection[J]. Science, 2003, 300(5616):65-67.
[14] Mekahli D, Bultynck G, Parys JB, et al. Endoplasmic-reticulum calcium depletion and disease[J]. Cold Spring Harb Perspect Biol, 2011, 3(6):a004317.

[1]	艾多, 徐安琦, 孔德钦, 刘颖, 于卫华, 王钊, 彭洁, 刘瑞, 张晓迪, 海春旭, 李文丽, 王欣, 刘江正. 催化性抗氧化剂AEOL-10150对氮芥诱导小鼠急性肝损伤的保护作用[J]. 癌变·畸变·突变, 2022, 34(1): 40-46,61.
[2]	尹亚萍, 王宇, 田雨阳, 马焜, 李百祥. 褪黑素在帕金森病治疗中的研究进展[J]. 癌变·畸变·突变, 2022, 34(1): 75-78,81.
[3]	曹家豪, 彭越, 段佳琪, 刘桥松, 刘启玲, 秦绪军. 酒精性脂肪肝的线粒体作用机制研究进展[J]. 癌变·畸变·突变, 2021, 33(6): 475-481.
[4]	李宏伟, 孔德钦, 于卫华, 吴昊, 王钊, 刘瑞, 海春旭, 王欣, 刘江正, 李文丽. 齐墩果酸对酒精诱导的大鼠胃壁氧化损伤的保护作用[J]. 癌变·畸变·突变, 2021, 33(5): 365-369.
[5]	蒋杉, 杜凤, 康秉文, 殷草草, 周健, 石莹, 王玥, 周耿瑶, 秦绪军. 异鼠李素通过减轻氧化应激延缓D-半乳糖诱导的人脐静脉内皮细胞衰老[J]. 癌变·畸变·突变, 2021, 33(2): 95-100.
[6]	赵俊伟, 华辰凤, 尚平平, 谢复炜, 李翔. 亚砷酸As (Ⅲ)诱导人肺癌A549细胞氧化应激反应的实验研究[J]. 癌变·畸变·突变, 2021, 33(1): 28-31.
[7]	郭茜晨, 张婷婷, 王润新, 赵法明, 盛夏. 内质网应激反应IRE1α信号通路与细胞衰老在癌症中的作用及关系的研究进展[J]. 癌变·畸变·突变, 2021, 33(1): 72-76.
[8]	张力, 张爱华, 王文娟. 甲基汞暴露致学习记忆功能障碍作用机制的研究进展[J]. 癌变·畸变·突变, 2020, 32(6): 490-493.
[9]	孙娇娇, 车碧众, 罗秋林, 翟兵中, 信丽丽. 1,25-二羟基维生素D₃对PM_2.5所致HBE细胞氧化损伤的保护作用[J]. 癌变·畸变·突变, 2020, 32(2): 92-97.
[10]	郭倪君, 林忠宁, 林育纯. 铁死亡信号介导抗肿瘤作用的细胞器调控机制研究进展[J]. 癌变·畸变·突变, 2020, 32(2): 149-154.
[11]	金磊, 周珈右, 韩家诚, 管浩军, 王帅, 彭洁, 王欣, 海春旭, 李文丽. 降脂酮对棕榈酸诱导BRL-3A细胞胰岛素抵抗的保护作用[J]. 癌变·畸变·突变, 2020, 32(1): 21-28.
[12]	王倩, 黄云飞, 雷磊, 苏德奇. 香烟烟雾与慢性应激联合暴露对大鼠学习记忆能力的影响[J]. 癌变·畸变·突变, 2019, 31(6): 474-478,482.
[13]	张旭光, 杜瑞琥, 陈彦平, 赵艳. 维生素E琥珀酸酯通过酸性神经磷脂酶-神经酰胺诱导胃癌细胞凋亡的机制研究[J]. 癌变·畸变·突变, 2019, 31(5): 379-384.
[14]	廖鼐, 龙子, 海春旭, 王欣. 2,3,7,8-四氯二苯并二噁英染毒或联合高脂饮食致小鼠糖脂代谢紊乱的实验研究[J]. 癌变·畸变·突变, 2019, 31(2): 111-118.
[15]	侯腾, 马璐, 张爱华. 组蛋白修饰在砷中毒发病机制中的研究进展[J]. 癌变·畸变·突变, 2019, 31(2): 153-156.

AMPK活性对HeLa细胞内质网功能稳态的影响

Effects of AMP-activated protein kinase activation on homeostasis of endoplasmic reticulum function in HeLa cells

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价