LncRNA ABHD11-AS1在Erastin诱导的胰腺癌细胞铁死亡中的作用及其机制

摘要
图/表
参考文献(0)
相关文章 (11)

全文: PDF (1613 KB) HTML (1 KB)
输出: BibTeX | EndNote (RIS) 背景资料

文章导读

摘要［摘要］目的：探讨长链非编码RNA(long noncoding RNA,lncRNA)α/β水解酶结构域蛋白11的反义链1(α/βhydrolase domain containing protein 11antisense strand 1，ABHD11AS1)在Erastin诱导的胰腺癌细胞铁死亡中的作用及其潜在机制。方法: 通过GEPIA平台分析ABHD11AS1在胰腺癌中的表达和患者预后。采用荧光实时定量PCR(qRTPCR)检测人胰腺癌PANC1、MIApaca2和PaTu8988细胞中ABHD11AS1表达，CCK8法计算Erastin半数抑制浓度(IC50)。在PANC1细胞中过表达ABHD11AS1，分别转染pcDNA3.1、pcDNA3.1ABHD11AS1质粒；在PaTu8988细胞中干扰ABHD11AS1表达，分别转染siControl、siRNA1、siRNA2，qRTPCR检测转染效率，分别予以对照(0 μmol/L Erastin)、Erastin(IC50浓度)处理，CCK8法检测细胞活性，ELISA法检测丙二醛和还原型谷胱甘肽(glutathione，GSH)含量；另在pcDNA3-1-ABHD11-AS1和siRNA1组加入Erastin的同时加入铁死亡挽救剂Ferrostatin1，坏死挽救剂Necrostatin-1或凋亡挽救剂Z-VAD-FMK，CCK8法检测细胞活性；免疫印迹法检测5组胰腺癌细胞中谷胱甘肽过氧化物酶4(glutathione peroxidase 4，GPX4)、雷帕霉素靶蛋白(mammalian target of rapamycin，mTOR)以及磷酸化mTOR(p-mTOR)表达水平。结果: 胰腺癌组织中ABHD11-AS1表达水平明显高于癌旁组织，高表达ABHD11-AS1组患者预后较差(P<0.05)。ABHD11-AS1相对表达量由低到高依次是人胰腺癌PANC1、MIApaca2和PaTu8988细胞，Erasin IC50趋势与ABHD11AS1表达量相同，依次是2.245、11.760、17.120 μmol/L。与pcDNA3.1组相比，pcDNA3.1ABHD11AS1组细胞活性及GSH含量明显增高，而丙二醛含量明显降低(P<0.05)；与siControl组相比，siRNA1组和siRNA2组细胞活性及GSH含量明显降低，丙二醛含量明显增高(P均<0.05)。仅Ferrostatin-1可以挽救pcDNA3.1-ABHD11AS1和siRNA1组细胞活性(P<0.05)。过表达ABHD11AS1可促进mTOR活化，增强GPX4表达(P<0.05)；而干扰ABHD11AS1则相反。结论: LncRNA ABHD11AS1可通过活化mTOR促进GPX4表达，从而促进胰腺癌细胞铁死亡抵抗。

	服务

	把本文推荐给朋友
	加入我的书架
	加入引用管理器
	E-mail Alert
	RSS
	作者相关文章

关键词 ：长链非编码RNAfont-family: ????, font-weight: 700, text-align: -webkit-right, ')" href="#">background-color: rgb(212, 226, 255);">, α/β-水解酶结构域蛋白11的反义链1(ABHD11-AS1)font-family: ????, font-weight: 700, text-align: -webkit-right, ')" href="#">background-color: rgb(212, 226, 255);">, 胰腺癌font-family: ????, font-weight: 700, text-align: -webkit-right, ')" href="#">background-color: rgb(212, 226, 255);">, 铁死亡font-family: ????, font-weight: 700, text-align: -webkit-right, ')" href="#">background-color: rgb(212, 226, 255);">, Erastin

Abstract：［Abstract］Objective： To investigate the role of long non-coding RNA (lncRNA) α/β-hydrolase domain containing protein 11-antisense strand 1 (ABHD11-AS1) in Erastin-induced ferroptosis of pancreatic cancer cells and its potential mechanism. Methods： The expression of ABHD11-AS1 and prognosis of patients in pancreatic cancer were analyzed by GEPIA platform. qRT-PCR was used to detect ABHD11-AS1 expression in human pancreatic cancer cells PANC1, MIApaca2 and PaTu8988, and CCK8 assay was used to calculate the IC50 of Erastin. PANC1 cells was transfected with pcDNA3.1 or PCDNA3.1-ABHD11-AS1 plasmid to overexpress ABHD11-AS1. PaTu8988 cells was transfected with siControl, siRNA1 and siRNA2 to interfere ABHD11-AS1; and the transfection efficiency was detected by qRT-PCR. The cells were treated with control (0 μmol/L Erastin), Erastin (IC50 concentration), respectively; and cell viability was detected by CCK8 assay. The content of malondialdehyde (MDA) and glutathione (GSH) were determined by ELISA. In addition, in pcDNA3.1ABHD11-AS1 and siRNA1 groups, Erastin was added and ferroptosis inhibiting agent Ferrostatin-1, necrosis salvage agent Necrosstatin-1 or apoptosis inhibitor ZVADFMK were added at the same time, and cell viability was detected by CCK8 assay. Western blotting was used to detect the protein expression of glutathione peroxidase 4 (GPX4), mammalian target of rapamycin (mTOR), and phosphorylated mTOR (p-mTOR) in pancreatic cancer cells of 5 groups. Results： The expression level of ABHD11-AS1 in pancreatic cancer tissues was significantly higher than that in adjacent tissues, and the prognosis of patients with higher expression of ABHD11-AS1 was poorer (P<0.05). The expression level of ABHD11-AS1 from low to high in human pancreatic cancer cells were PANC1, MIApaca-2, and PaTu8988. The trend of Erasin IC50 was the same as that of ABHD11-AS1, which was 2.245, 11.760, 17.120 μmol/L, respectively. Compared with the pcDNA3.1 group, PCDNA3.1ABHD11AS1 group showed higher levels of cell viability and GSH content, while lower content of MDA (all P<0.05). Compared with the siControl group, the cell viability and GSH content in siRNA1 group and siRNA2 group decreased significantly, and the MDA content increased significantly (P<0.05). Only Ferrostatin-1 could rescue the cell viability of pcDNA3.1ABHD11-AS1 and siRNA1 groups (P<0.05). ABHD11-AS1 increased cell viability under Erastin treatment by enhancing ferroptosis resistance. Overexpression of ABHD11-AS1 could promote the activation of mTOR and enhance the expression of GPX4 (P<0.05); knockdown of ABHD11-AS1 produced the opposite effect. Conclusion： LncRNA ABHD11-AS1 could promote the expression of GPX4 by activating mTOR1, thus promoting ferroptosis resistance of pancreatic cancer cells.

Key words：

收稿日期: 2022-02-24

基金资助: 国家自然科学基金面上项目(82072754)；江苏省卫生健康委科技项目(M2020011)；江苏省社会发展基金资助项目(BE2018689)；镇江市社会发展项目(SH2018033)

通讯作者: 徐岷(通讯作者)，主任医师，博士生导师，E-mail: peterxu1974@163.com

作者简介: 徐春晖(1993—)，男，硕士研究生；

引用本文:

徐春晖，王慧之，刘雅雯，等.. LncRNA ABHD11-AS1在Erastin诱导的胰腺癌细胞铁死亡中的作用及其机制[J]. 江苏大学学报:医学版, 2023, 33(01): 1-8. XU Chunhui1, WANG Huizhi1, LIU Yawen1, LI Zheng1, GONG Aihua2, XU Min1. Role of LncRNA ABHD11AS1 in Erastininduced ferroptosis of pancreatic cancer cells and its mechanism. Journal of Jiangsu University(Medicine Edition), 2023, 33(01): 1-8.

链接本文:

http://zzs.ujs.edu.cn/xbyxb/CN/ 或 http://zzs.ujs.edu.cn/xbyxb/CN/Y2023/V33/I01/1

［1］Siegel RL, Miller KD, Fuchs HE, et al. Cancer Statistics, 2021［J］. CA Cancer J Clin, 2021, 71(1): 7-33.

［2］虞先濬, 刘亮, 徐华祥, 等. 胰腺癌综合诊治指南(2018版)［J］. 临床肝胆病杂志, 2018, 34(10): 2109-2120.

［3］Wang C, Huang H. Effects of ribosomal associated protein hnRNP D in gemcitabineresistant pancreatic cancer［J］. Biochem Biophys Res Commun, 2021, 579: 181-187.

［4］Palazzo AF, Koonin EV. Unctional long noncoding RNAs evolve from junk transcripts［J］. Cell, 2020, 183(5): 1151-1161.

［5］Liu SJ, Dang HX, Lim DA, et al. Long noncoding RNAs in cancer metastasis［J］. Nat Rev Cancer, 2021, 21(7): 446-460.

［6］Liu Y, Feng W, Liu W, et al. Circulating lncRNA ABHD11AS1 serves as a biomarker for early pancreatic cancer diagnosis［J］. J Cancer, 2019, 10(16): 3746-3756.

［7］Luo J, Jiang Y, Wu L, et al. Long noncoding RNA ABHD11AS1 promotes colorectal cancer progression and invasion through targeting the integrin subunit α 5/focal adhesion kinase/phosphoinositide 3 kinase/Akt signaling pathway［J］. Aging (Albany NY), 2021, 13(16): 20179-20191.

［8］Zhang W, Huang X, Shi J. EZH2mediated lncRNA ABHD11AS1 promoter regulates the progression of ovarian cancer by targeting miR133a3p［J］. Anticancer Drugs, 2021, 32(3): 269-277.

［9］Dixon SJ, Lemberg KM, Lamprecht MR, et al. Ferroptosis: an irondependent form of nonapoptotic cell death［J］. Cell, 2012, 149(5): 1060-1072.

［10］Distefano AM, Lopez GA, Setzes N, et al. Ferroptosis in plants: triggers, proposed mechanisms, and the role of iron in modulating cell death［J］. J Exp Bot, 2021, 72(6): 2125-2135.

［11］Bogacz M, KrauthSiegel RL. Tryparedoxin peroxidasedeficiency commits trypanosomes to ferroptosistype cell death［J］. Elife, 2018, 7: e37503.

［12］Ye Z, Hu Q, Zhuo Q, et al. Abrogation of ARF6 promotes RSL3induced ferroptosis and mitigates gemcitabine resistance in pancreatic cancer cells［J］. Am J Cancer Res, 2020, 10(4): 1182-1193.

［13］Zhang Y, Tan H, Daniels JD, et al. Imidazole ketone erastin induces ferroptosis and slows tumor growth in a mouse lymphoma model［J］. Cell Chem Biol, 2019, 26(5): 623-633.e9.

［14］Viswanathan VS, Ryan MJ, Dhruv HD, et al. Dependency of a therapyresistant state of cancer cells on a lipid peroxidase pathway［J］. Nature, 2017, 547(7664): 453-457.

［15］Zuo YB, Zhang YF, Zhang R, et al. Ferroptosis in cancer progression: role of noncoding RNAs［J］. Int J Biol Sci, 2022, 18(5): 1829-1843.

［16］Chen X, Kang R, Kroemer G, et al. Broadening horizons: the role of ferroptosis in cancer［J］. Nat Rev Clin Oncol, 2021, 18(5): 280-296.

［17］Wang H, Liu C, Zhao YX, et al. Inhibition of LONP1 protects against erastininduced ferroptosis in pancreatic ductal adenocarcinoma PANC1 cells［J］. Biochem Biophys Res Commun, 2020, 522(6): 1063-1068.

［18］Daher B, Parks SK, Durivault J, et al. Genetic ablation of the cystine transporter xCT in PDAC cells inhibits mTORC1, growth, survival, and tumor formation via nutrient and oxidative stresses［J］. Cancer Res, 2019, 79(15): 3877-3890.

［19］蔡晓杰, 王鸣, 高洁, 等. 二甲双胍逆转低氧诱导的胰腺癌细胞铁死亡抵抗及潜在的作用机制［J］. 江苏大学学报(医学版), 2020, 30(2): 125-130.

［20］Xia J, Song XY, Meng J, et al. Endothelial progenitor cellsderived exosomes transfer microRNA30e5p to regulate Erastininduced ferroptosis in human umbilical vein endothelial cells via the specificity protein 1/adenosine monophosphateactivated protein kinase axis［J］. Bioengineered, 2022, 13(2): 3566-3580.

［21］Li Z, Dai Hui Q, Huang X, et al. Artesunate synergizes with sorafenib to induce ferroptosis in hepatocellular carcinoma［J］. Acta Pharmacol Sin, 2021, 42(2): 301-310.

［22］Kim EH, Shin D, Lee J, et al. CISD2 inhibition overcomes resistance to sulfasalazineinduced ferroptotic cell death in head and neck cancer［J］. Cancer Lett, 2018, 432: 180-190.

［23］Yao X, Xie R, Cao Y, et al. Simvastatin induced ferroptosis for triplenegative breast cancer therapy［J］. J Nanobiotechnology, 2021, 19(1): 311.

［24］Zhu S, Yu Q, Huo C, et al. Ferroptosis: a novel mechanism of artemisinin and its derivatives in cancer therapy［J］. Curr Med Chem, 2021, 28(2): 329-345.

［25］Yang WS, SriRamaratnam R, Welsch ME, et al. Regulation of ferroptotic cancer cell death by GPX4［J］. Cell, 2014, 156(1-2): 317-331.