[1]付永生a,卢静芬a,赵 昕b,等.DNER 通过抑制线粒体自噬促进胃癌细胞恶性进展的机制研究[J].现代检验医学杂志,2024,39(04):50-55.[doi:10.3969/j.issn.1671-7414.2024.04.010]
　FU Yongshenga,LU Jingfena,ZHAO Xinb,et al.Study on the Mechanism of DNER Promoting Malignant Progression of Gastric Cancer Cells by Inhibiting Mitochondrial Autophagy[J].Journal of Modern Laboratory Medicine,2024,39(04):50-55.[doi:10.3969/j.issn.1671-7414.2024.04.010]
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DNER 通过抑制线粒体自噬促进胃癌细胞恶性进展的机制研究()
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《现代检验医学杂志》[ISSN:/CN:]

卷:
第39卷
期数:
2024年04期
页码:
50-55
栏目:
论著
出版日期:
2024-07-15

文章信息/Info

Title:
Study on the Mechanism of DNER Promoting Malignant Progression of Gastric Cancer Cells by Inhibiting Mitochondrial Autophagy
文章编号:
1671-7414（2024）04-050-06
作者:
付永生1a; 卢静芬1a; 赵 昕1b; 王 卫2; 朱其聪1a
（1. 解放军联勤保障部队第九二八医院 a. 血液肿瘤科；b. 普通外科，海口 570100；2. 海口市第三人民医院普通外科，海口 570100）
Author(s):
FU Yongsheng1a; LU Jingfen1a; ZHAO Xin1b; WANG Wei2; ZHU Qicong1a
（1a. Department of Hematology and Oncology；1b. Department of General Surgery，928 Medical Hospital of PLA Joint Logistic Support Force，Haikou 570100, China；2. Department of General Surgery，the Third People’s Hospital of Haikou,Haikou 570100, China）
关键词:
胃癌; 自噬; Delta/Notch 样表皮生长因子相关受体; 线粒体融合; 线粒体裂变
分类号:
R735.2；R730.43
DOI:
10.3969/j.issn.1671-7414.2024.04.010
文献标志码:
A
摘要:
目的 研究Delta/Notch 样表皮生长因子相关受体（delta/notch-like epidermal growth factor-related receptor，DNER）在胃癌中的作用及其调节机制。方法 通过实时定量聚合酶链反应（qRT-PCR）和蛋白免疫印迹（Westernblot）检测胃癌组织和细胞中DNER mRNA 和蛋白表达水平。构建沉默DNER 表达的胃癌细胞系SGC7901，用线粒体动力相关蛋白（dynamin-related protein 1，DRP1）抑制剂Mdivi-1 处理细胞。CCK-8 法、Transwell 实验和流式细胞术分别检测细胞活力、侵袭能力和细胞凋亡。Western blot 检测DNER 蛋白、凋亡相关蛋白[ 半胱天冬氨酸蛋白酶3（cysteinylaspartate-specific proteinase -3，Caspase-3）、Bcl-2 相关X 蛋白（Bcl-2 associated X，Bax）]、自噬相关蛋白[ 微管相关蛋白1 轻链3- Ⅱ / Ⅰ（microtubule-associated protein 1 light chain 3-II/ Ⅰ，LC3 Ⅱ / Ⅰ）、p62，PTEN 诱导激酶1（PTENinduced putative kinase 1，PINK1）和Parkin]，以及线粒体裂变和融合蛋白[DRP1，线粒体分裂因子（mitochondrialfission factor，MFF）、线粒体分裂蛋白1(fission mitochondrial 1, FIS1)、视神经萎缩蛋白1（optic atrophy 1，OPA1）、线粒体融合蛋白1(mitofusin 1，MFN1) 和MFN2] 水平。结果 胃癌肿瘤组织和细胞中DNER mRNA，蛋白表达水平分别显著高于相邻正常组织（t=-52.485，-46.955）和人正常胃上皮细胞（F=60.551，60.652），差异具有统计学意义（均P<0.001）。沉默DNER 显著抑制SGC7901 细胞的增殖和侵袭、诱导细胞凋亡、增加细胞凋亡相关蛋白表达，差异具有统计学意义（t=8.026~25.903，均P<0.05）。沉默DNER显著升高LC3Ⅱ / Ⅰ比率（t=18.086），降低p62 蛋白水平（t=6.747），促进PINK1和Parkin 蛋白在线粒体的聚集（t=15.630，18.171），抑制线粒体融合蛋白OPA1，MFN1和MFN2表达（t=12.835，8.963，9.732），促进线粒体裂变蛋白DRP1，MFF 和FIS1 表达（t=16.034，16.939，15.971），差异具有统计学意义（均P<0.05）。Mdivi-1 处理可抵消沉默DNER 对胃癌细胞线粒体自噬及细胞增殖、侵袭和凋亡的影响。结论 DNER 通过抑制线粒体动力学失衡减少线粒体自噬，促进细胞增殖和侵袭，抑制细胞凋亡，从而促进胃癌进展。
Abstract:
Objective To investigate the role of delta/notch-like epidermal growth factor-related receptor (DNER) in gastric cancer and its regulatory mechanism. Methods The mRNA and protein levels of DNER in gastric cancer tissues and cells were detected with quantitative real time polymerase chain reaction (qRT-PCR) and Western blot. Gastric cancer cell line SGC7901 with silenced DNER expression was constructed, and cells were treated with mitochondrial dynamin-related protein 1 (DRP1) inhibitor Mdivi-1. CCK-8 assay, Transwell assay, and flow cytometry were used to detect cell viability, invasion ability and apoptosis, respectively. Western blot was used to detect DNER protein levels, apoptosis-associated proteins [Cysteinyl aspartatespecific proteinase-3 (Caspase-3), Bcl-2 Associated X (Bax)], autophagy associated proteins [microtubule-associated protein 1 light chain 3-II/ Ⅰ , LC3 Ⅱ / Ⅰ ), p62, PTEN induced putative kinase 1 (PINK1) and Parkin], and mitochondrial fission and fusion protein [DRP1, mitochondrial fission factor (MFF), mitochondrial fission protein 1(FIS1), Optic Atrophy 1(OPA1), mitofusin 1 (MFN1) and MFN2] levels. Results The expression levels of DNER mRNA and protein in gastric cancer tissues were higher than those in adjacent normal tissues(t=-52.485, -46.955), while expression levels of DNER mRNA and protein in gastric cancer cells were higher than those in normal gastric epithelial cells(F=60.551, 60.652), and the differences were significant (P<0.001). Silencing DNER inhibited the proliferation and invasion of SGC7901 cells, induced apoptosis, and increased the expression of apoptosis-related proteins, with significant differences (t= 8.026 ～ 25.903, all P<0.05). Silenced DNER increased LC3 Ⅱ / Ⅰ ratio (t=18.086), decreased p62 protein level (t=6.747), promoted the aggregation of PINK1 and Parkin proteins in mitochondria (t=15.630, 18.171), inhibited the expression of mitochondrial fusion proteins OPA1, MFN1 and MFN2 (t=12.835, 8.963, 9.732), and promoted the expression of mitochondrial fission proteins DRP1, MFF and FIS1 (t=16.034, 16.939, 15.971), with significant differences (all P<0.05). Mdivi-1 treatment could counteract the effects of silencing DNER on mitochondrial autophagy, proliferation, invasion and apoptosis of gastric cancer cells. Conclusion DNER can reduce mitochondrial autophagy by inhibiting mitochondrial dynamic imbalance, promote cell proliferation and invasion, and inhibit cell apoptosis, thus promoting the progression of gastric cancer.

参考文献/References:

[1] HUANG R J, LASZKOWSKA M, IN H, et al. Controlling gastric cancer in a world of heterogeneous risk[J]. Gastroenterology, 2023, 164(5): 736-751.
[2] GUAN Wenlong, HE Ye, XU Ruihua. Gastric cancer treatment: recent progress and future perspectives[J]. Journal of Hematology & Oncology, 2023, 16(1): 57.
[3] FERRO F, SERVAIS S, BESSON P, et al. Autophagy and mitophagy in cancer metabolic remodelling [J]. Seminars in Cell Developmental Biology, 2020, 98: 129-138.
[4] PICCA A, FAITG J, AUWERX J, et al. Mitophagy in human health, ageing and disease [J]. Nature Metabolism, 2023, 5(12): 2047-2061.
[5] LIU Lei, LI Yanjun, CHEN Guo, et al. Crosstalk between mitochondrial biogenesis and mitophagy to maintain mitochondrial homeostasis[J]. Journal of Biomedical Science, 2023, 30(1): 86.
[6] 代晶, 张勇, 潘长清, 等．RMP 通过AMPK 通路调节线粒体稳态和氧化应激诱导卵巢癌细胞增殖与凋亡的机制研究[J]．现代检验医学杂志, 2023, 38(2):57-62. DAI Jing, ZHANG Yong, PAN Changqing, et al. Mechanism of RMP regulating mitochondrial homeostasis and oxidative stress-induced ovarian cancer cell proliferation and apoptosis through the AMPK pathway [J]. Journal of Modern Laboratory Medicine, 2023, 38(2): 57-62.
[7] RADEMAKER G, BOUMAHD Y, PEIFFER R, et al. Myoferlin targeting triggers mitophagy and primes ferroptosis in pancreatic cancer cells [J]. Redox Biology, 2022, 53: 102324.
[8] MENG Yang, QIU Lei, ZENG Xinyi, et al. Targeting CRL4 suppresses chemoresistant ovarian cancer growth by inducing mitophagy[J]. Signal Transduction and Targeted therapy, 2022, 7(1): 388.
[9] SUN Yueqin, SHEN Weitao, HU Shulu, et al. METTL3 promotes chemoresistance in small cell lung cancer by inducing mitophagy[J]. Journal of Experimental & Clinical Cancer Research, 2023, 42(1): 65.
[10] DENG Rong, ZHANG Hailiang, HUANG Junhao, et al. MAPK1/3 kinase-dependent ULK1 degradation attenuates mitophagy and promotes breast cancer bone metastasis[J]. Autophagy, 2021, 17(10): 3011-3029.
[11] WANG Xiaohong, ZHANG Lianhai, CHAN F K L, et al. Gamma-glutamyltransferase 7 suppresses gastric cancer by cooperating with RAB7 to induce mitophagy[J]. Oncogene, 2022, 41(26): 3485-3497.
[12] WANG Lijun, WU Qi, LI Zhiyu, et al. Delta/notchlike epidermal growth factor-related receptor promotes stemness to facilitate breast cancer progression [J]. Cellular Signalling, 2019, 63: 109389.
[13] WANG Zhong, LI Zhiyu, WU Qi, et al. DNER promotes epithelial-mesenchymal transition and prevents chemosensitivity through the Wnt/β-catenin pathway in breast cancer [J]. Cell Death Disease, 2020,11(8): 642.
[14] WANG Lijun, WU Qi, ZHU Shan, et al. Delta/notchlike epidermal growth factor-related receptor (DNER) orchestrates stemness and cancer progression in prostate cancer [J]. American Journal of Translational Research, 2017, 9(11): 5031-5039.
[15] HE Rui, ZHANG Wunai, CHEN Shuo, et al. Transcriptional profiling reveals the regulatory role of DNER in promoting pancreatic neuroendocrine neoplasms [J]. Frontiers in Genetics, 2020, 11: 587402.
[16] LIANG Yan, LUO Hesheng, ZHANG Haidong, et al. Oncogene delta/Notch-like EGF-related receptor promotes cell proliferation, invasion, and migration in hepatocellular carcinoma and predicts a poor prognosis[J]. Cancer Biotherapy and Radiopharmaceuticals, 2018, 33(9): 380-386.
[17] WANG Qun, LI Yaqiong, LI Jiamei, et al. Delta and notch-like epidermal growth factor-related receptor suppresses human glioma growth by inhibiting oncogene TOR4A[J]. Journal of Cancer Research and Therapeutics, 2022, 18(5): 1372-1379.
[18] TO H T N, PARK J H, KIM J W, et al. Delta/notch-like epidermal growth factor-related receptor (DNER), a potential prognostic marker of gastric cancer regulates cell survival and cell cycle progression[J]. International Journal of Molecular Sciences, 2023, 24(12): 10077.
[19] YANG Wenjuan, ZHAO Heping, YU Yan, et al. Updates on global epidemiology, risk and prognostic factors of gastric cancer[J]. World Journal of Gastroenterology, 2023, 29(16): 2452-2468.
[20] NG M Y W, WAI T, SIMONSEN A. Quality control of the mitochondrion[J]. Developmental Cell, 2021, 56(7):881-905.
[21] 骆慧, 于月, 廖照辉, 等. 线粒体自噬的信号通路及其在肿瘤中的研究进展[J]. 临床肿瘤学杂志, 2022,27(7): 661-668． LUO Hui, YU Yue, LIAO Zhaohui, et al. Progress on the signal pathway of mitochondrial autophagy and its application in tumor [J]. Chinese Clinical Oncology, 2022, 27(7): 661-668.
[22] ADEBAYO M, SINGH S, SINGH A P, et al. Mitochondrial fusion and fission: the fine-tune balance for cellular homeostasis[J]. FASEB Journal, 2021, 35(6): e21620.
[23] ZHANG Baoyi, PAN Cunyao, FENG Chong, et al. Role of mitochondrial reactive oxygen species in homeostasis regulation[J]. Redox Report, 2022, 27(1): 45-52.
[24] ROTH K G, MAMBETSARIEV I, KULKARNI P, et al. The mitochondrion as an emerging therapeutic target in cancer[J]. Trends in Molecular Medicine, 2020, 26(1): 119-134.
[25] SREEDHAR A, AGUILERA-AGUIRRE L, SINGH K K. Mitochondria in skin health, aging, and disease [J]. Cell death Disease, 2020, 11(6): 444.
[26] 陈元菁, 艾红军, 王湘云, 等．COPD 患者线粒体功能及代谢水平变化检测与骨骼肌功能障碍的相关性研究[J]. 现代检验医学杂志, 2020, 35(6): 25-27, 32. CHEN Yuanjing, AI Hongjun, WANG Xiangyun, et al. Study on the correlation between changes detection in mitochondrial function, metabolic level and skeletal muscle dysfunction in patients with COPD [J]. Journal of Modern Laboratory Medicine, 2020, 35(6): 25-27, 32.
[27] JIAO Haifeng, JIANG Dong, HU Xiaoyu, et al. Mitocytosis, a migrasome-mediated mitochondrial quality-control process[J]. Cell, 2021, 184(11): 2896-2910, e13.
[28] POPOV L D. Mitochondrial biogenesis: an update[J]. Journal of Cellular and Molecular Medicine, 2020, 24(9): 4892-4899.

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备注/Memo

备注/Memo:
基金项目：海南省卫生厅科学研究项目（编号：22A200057）。
作者简介：付永生（1985-），男，本科，主治医师，研究方向：肿瘤学，E-mail：FYs850606@126.com。
通讯作者：朱其聪（1973-），男，硕士研究生，副主任医师，研究方向：肿瘤学，E-mail： zhuqico1234@163.com。
更新日期/Last Update: 2024-07-15