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《天津医科大学学报》[ISSN:1006-8147/CN:12-1259/R]

卷:

30卷

期数:

2024年02期

页码:

110-115

栏目:

肿瘤疾病专题

出版日期:

2024-03-20

文章信息/Info

Title:

The mechanism of telomerase TERT resisting ferroptosis in gastric cancer cells by translocating to mitochondria

文章编号:

1006-8147（2024）02-0110-06

作者:

刘思琪; 郝名英; 黄欣宇; 耿 鑫

（天津医科大学基础医学院生物化学与分子生物学系，天津 300070）

Author(s):

LIU Siqi; HAO Mingying; HUANG Xinyu; GENG Xin

（Department of Biochemistry and Molecular Biology，School of Basic Medical Sciences，Tianjin Medical University，Tianjin 300070，China）

关键词:

端粒酶; 胃癌; 铁死亡; 线粒体

Keywords:

telomerase; gastric cancer; ferroptosis; mitochondria

分类号:

R735.2

DOI:

10.20135/j.issn.1006-8147.2024.02.0110

文献标志码:

A

摘要:

目的：探究胃癌细胞铁死亡发生时，端粒酶TERT如何抵抗铁死亡的发生。方法：提取胃癌细胞BGC823的线粒体及细胞总蛋白，通过蛋白质印迹实验检测10 μmol/L erastin处理细胞后，铁死亡相关蛋白SLC7A11、谷胱甘肽过氧化物酶4（GPX4）及端粒酶蛋白TERT表达量的变化以及线粒体中TERT含量变化；通过免疫荧光实验检测TERT是否定位于线粒体；使用5 μmol/L bosutinib预处理细胞后再加入10 μmol/L erastin处理细胞，通过免疫荧光实验检测TERT与线粒体定位情况；通过活性氧簇（ROS）检测试剂盒、丙二醛（MDA）检测试剂盒、Fe2+检测试剂盒分别检测ROS、MDA、Fe2+含量变化；通过CCK-8及划痕实验检测细胞生长、增殖及迁移能力。结果：蛋白质印迹实验结果显示，GPX4（t=15.30，P<0.001）、SLC7A11（t=5.228，P<0.01）、TERT（t=3.682，P<0.05）水平明显降低，线粒体中TERT含量降低（t=6.736，P<0.001）。免疫荧光实验结果显示，TERT定位于线粒体，联合用药组细胞免疫荧光实验结果显示TERT更多的定位在细胞核。联合用药组与单独用药组相比，铁死亡指标ROS（t=4.109，P<0.05）、MDA（t=6.491，P<0.01）、Fe2+（F=9.703，P<0.01）含量均升高，CCK-8（F=4.706，P<0.05）及划痕实验（t=4.631，P<0.05）结果显示细胞增殖、迁移能力进一步降低。结论：铁死亡发生时，TERT由细胞核向线粒体进行转位，使用bosutinib抑制TERT向线粒体转位后，铁死亡相关指标增强，抑制TERT转位可以增强铁死亡的发生，抑制胃癌细胞生长、增殖。

Abstract:

Objective：To explore how telomerase TERT resists ferroptosis in gastric cancer cells. Methods：Mitochondria and total cell proteins of gastric cancer cell BGC823 were extracted. Changes in the expression levels of ferroptosis related proteins SLC7A11，glutathione peroxidase 4（GPX4），and telomerase protein TERT after treatment with 10 μmol/L erastin in cells were detected by Western blotting. Whether TERT was localized in mitochondria was detected through immunofluorescence assay. TERT and mitochondrial localization were detected by immunofluorescence assay after 5 μmol/L bosutinib pretreatment and 10 μmol/L erastin treatment of cells. Changes in ROS，MDA，and Fe2+ contents were detected by reactive oxygen species（ROS） detection kit，malondialdehyde （MDA） detection kit，and Fe2+ detection kit，respectively. Cell growth，proliferation，and migration were measured by CCK-8 and wound healing.Results：Western blotting showed a significant decrease in GPX4（t=15.30，P<0.001），SLC7A11（t=5.228，P<0.01），and TERT（t=3.682， P<0.05） protein levels，and reduction in the content of TERT in mitochondria（t=6.736，P<0.001）. The immunofluorescence experiment results showed that TERT was localized in mitochondria，and the combined use of the drug group showed that TERT was more localized in the nucleus. Compared with the monotherapy group，the combination therapy group showed an increase in iron death markers ROS（t=4.109，P<0.05），MDA（t=6.491，P<0.01），and Fe2+（F=9.703，P<0.01） content. The CCK-8 and wound healing results showed a further decrease in cell proliferation and migration ability. Conclusion：When ferroptosis occurs，TERT translocates from the nucleus to mitochondria. Inhibiting TERT translocation to mitochondria with bosutinib enhances ferroptosis related indicators，indicating that inhibiting TERT translocation can enhance ferroptosis and inhibit the growth and proliferation of gastric cancer cells.

参考文献/References:

[1] BRAY F，FERLAY J，SOERJOMATARAM I，et al. Global cancer statistics 2018：GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries[J]. CA Cancer J Clin，2018，68（6）：394-424.
[2] SMYTH E C，NILSSON M，GRABSCH H I，et al. Gastric cancer[J]. Lancet，2020，396（10251）：635-648.
[3] JEONG Y S，EUN Y G，LEE S H，et al. Clinically conserved genomic subtypes of gastric adenocarcinoma[J]. Mol Cancer，2023，22（1）：147.
[4] ZHANG C，LIU X，JIN S，et al. Ferroptosis in cancer therapy：a novel approach to reversing drug resistance[J]. Mol Cancer，2022，21（1）：47.
[5] ZHENG J，CONRAD M. The metabolic underpinnings of ferropto-sis[J]. Cell Metab，2020，32（6）：920-937.
[6] ZHANG R，KANG R，TANG D. Ferroptosis in gastrointestinal cancer：from mechanisms to implications[J]. Cancer Lett，2023，561：216147.
[7] HAENDELER J，HOFFMANN J，BRANDES R P，et al. Hydrogen peroxide triggers nuclear export of telomerase reverse transcriptase via Src kinase family-dependent phosphorylation of tyrosine 707[J]. Mol Cell Biol，2003，23（13）：4598-4610.
[8] ROSEN J，JAKOBS P，ALE-AGHA N，et al. Non-canonical functions of telomerase reverse transcriptase-impact on redox homeostasis[J]. Redox Biol，2020，34：101543.
[9] LING X，WEN L，ZHOU Y. Role of mitochondrial translocation of telomerase in hepatocellular carcinoma cells with multidrug resistance[J]. Int J Med Sci，2012，9（7）：545-554.
[10] GAN B. Mitochondrial regulation of ferroptosis[J]. J Cell Biol，2021， 220（9）：e202105043.
[11] WU S，MAO C，KONDIPARTHI L，et al. A ferroptosis defense mecha-nism mediated by glycerol-3-phosphate dehydrogenase 2 in mitochondria[J]. Proc Natl Acad Sci U S A，2022，119（26）：e2121987119.
[12] MAO C，LIU X，ZHANG Y，et al. DHODH-mediated ferroptosis defence is a targetable vulnerability in cancer[J]. Nature，2021，593（7860）：586-590.
[13] QIU S，ZHONG X，MENG X，et al. Mitochondria-localized cGAS suppresses ferroptosis to promote cancer progression[J]. Cell Res，2023，33（4）：299-311.
[14] 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.
[15] DANG Q，SUN Z，WANG Y，et al. Ferroptosis：a double-edged sword mediating immune tolerance of cancer[J]. Cell Death Dis，2022，13（11）：925.
[16] PULIGA E，CORSO S，PIETRANTONIO F，et al. Microsatellite instability in gastric cancer：between lights and shadows[J]. Cancer Treat Rev，2021，95：102175.
[17] HAENDELER J，DROSE S，BUCHNER N，et al. Mitochondrial telomerase reverse transcriptase binds to and protects mitochondrial DNA and function from damage[J]. Arterioscler Thromb Vasc Biol，2009，29（6）：929-935.
[18] ARNDT G M，MACKENZIE K L. New prospects for targeting te-lomerase beyond the telomere[J]. Nat Rev Cancer，2016，16（8）：508-524.
[19] HEIDENREICH B，KUMAR R. TERT promoter mutations in telo-mere biology[J]. Mutat Res Rev Mutat Res，2017，771：15-31.
[20] INDRAN I R，HANDE M P，PERVAIZ S. hTERT overexpression alleviates intracellular ROS production，improves mitochondrial function，and inhibits ROS-mediated apoptosis in cancer cells[J]. Cancer Res，2011，71（1）：266-276.
[21] ABBAS R，HSYU P H. Clinical pharmacokinetics and pharmacodynamics of bosutinib[J]. Clin Pharmacokinet，2016，55（10）：1191-1204.
[22] CORTES J E，GAMBACORTI-PASSERINI C，DEININGER M W，et al. Bosutinib versus imatinib for newly diagnosed chronic myeloid leukemia：results from the randomized BFORE trial[J]. J Clin Oncol，2018，36（3）：231-237.
[23] MIWA S，CZAPIEWSKI R，WAN T，et al. Decreased mTOR signalling reduces mitochondrial ROS in brain via accumulation of the telomerase protein TERT within mitochondria[J]. Aging （Albany NY），2016，8（10）：2551-2567.
[24] PATRICK S，GOWDA P，LATHORIA K，et al. YAP1-mediated regulation of mitochondrial dynamics in IDH1 mutant gliomas[J]. J Cell Sci， 2021， 134（22）：jcs259188.

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备注/Memo

备注/Memo:

基金项目 天津市教委科研计划（2021ZD037）
作者简介 刘思琪（1998-），女，硕士在读，研究方向：医学生物化学与分子生物学；通信作者 ：耿鑫，E-mail：gengx@tmu.edu.cn。

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更新日期/Last Update: 2024-03-20