[1]王敬如,张 琳,李峰敏.PLA2G4C通过p38/MAPK信号通路介导线粒体自噬促进DLBCL进展的实验研究[J].现代检验医学杂志,2024,39(06):61-66.[doi:10.3969/j.issn.1671-7414.2024.06.010]
　WANG Jingru,ZHANG Lin,LI Fengmin.Experimental Study on the Mechanism of Mitochondrial Autophagy Promoted DLBCL Progression Mediated by PLA2G4C Via p38/MAPK Signaling Pathway[J].Journal of Modern Laboratory Medicine,2024,39(06):61-66.[doi:10.3969/j.issn.1671-7414.2024.06.010]
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PLA2G4C通过p38/MAPK信号通路介导线粒体自噬促进DLBCL进展的实验研究()

《现代检验医学杂志》[ISSN:/CN:]

卷:
第39卷
期数:
2024年06期
页码:
61-66
栏目:
论著
出版日期:
2024-11-15

文章信息/Info

Title:
Experimental Study on the Mechanism of Mitochondrial Autophagy Promoted DLBCL Progression Mediated by PLA2G4C Via p38/MAPK Signaling Pathway
文章编号:
1671-7414（2024）06-061-06
作者:
王敬如，张 琳，李峰敏
（秦皇岛市第一医院血液内科，河北秦皇岛 066000）
Author(s):
WANG Jingru，ZHANG Lin，LI Fengmin
（Department of Hematology, the First Hospital of Qinhuangdao, Hebei Qinhuangdao 066000, China）
关键词:
弥漫性大B 细胞淋巴瘤; 磷脂酶A2 Ⅳ C 组; 线粒体自噬; p38/MAPK 信号通路
分类号:
R446.8
DOI:
10.3969/j.issn.1671-7414.2024.06.010
文献标志码:
A
摘要:
目的 研究磷脂酶A2 Ⅳ C 组（phospholipase A2 group Ⅳ C，PLA2G4C）在弥漫性大B 细胞淋巴瘤（diffuselarge B-cell lymphoma，DLBCL）中的作用及其可能调节机制。方法 通过免疫印迹法（Western blot）检测PLA2G4C在DLBCL组织和细胞中的表达。构建PLA2G4C过表达或敲低表达的DLBCL细胞系，后用自噬抑制剂氯喹（Chloroquine，CQ）或p38 抑制剂SB203580 处理细胞24h。实时定量聚合酶链反应（qRT-PCR）检测PLA2G4C 转染效率；Westernblot 检测PLA2G4C 蛋白、线粒体自噬相关蛋白[ 微管相关蛋白1 轻链3- Ⅱ / Ⅰ（microtubule-associated protein 1 lightchain 3- Ⅱ / Ⅰ，MAP1 LC3 Ⅱ / Ⅰ ），Beclin1，p62，PTEN 诱导激酶1（PTEN induced putative kinase 1，PINK1）和Parkin]，p38/ 丝裂原激活蛋白激酶（mitogen activated protein kinases，MAPK）通路相关蛋白[ 磷酸化p38/MAPK（phosphorylated-p38/MAPK，p-p38/MAPK）] 表达水平；CCK-8 法、Transwell 实验和流式细胞术分别检测细胞增殖、侵袭和凋亡能力。进一步构建异种移植瘤裸鼠模型，观察PLA2G4C 对裸鼠体内肿瘤生长及线粒体自噬的影响。结果 DLBCL 患者淋巴瘤组织中PLA2G4C 蛋白表达显著高于反应性增生淋巴结组织（3.47±0.42 vs 1.01±0.02），差异具有统计学意义（t= -37.002，P<0.001）；DLBCL 细胞中PLA2G4C 蛋白水平显著高于人淋巴母细胞样细胞系和B 细胞淋巴瘤细胞系，差异具有统计学意义（F=73.771，P<0.001）。沉默PLA2G4C 显著降低DLBCL 细胞活力和侵袭能力，诱导细胞凋亡（t=6.909 ～ 11.390）；过表达PLA2G4C 后结果与之相反（t=2.392~19.778），差异具有统计学意义（均P<0.001）。且过表达PLA2G4C 显著促进线粒体自噬的发生，而CQ 或SB203580 处理则可显著逆转PLA2G4C 过表达对DLBCL 细胞生物学行为及线粒体自噬的作用。体内裸鼠实验显示，敲低PLA2G4C 显著抑制移植瘤裸鼠体内肿瘤生长及线粒体自噬相关蛋白表达，差异具有统计学意义（t=13.816 ～ 25.926，均P<0.001）。结论 PLA2G4C 在DLBCL中表达显著上调，可能通过促进p38/MAPK 信号通路介导的线粒体自噬，促进肿瘤细胞增殖和侵袭，抑制细胞凋亡，参与DLBCL 的发生发展。
Abstract:
Objective To investigate the role of phospholipase A2 Group Ⅳ C (PLA2G4C) in diffuse large B-cell lymphoma (DLBCL) and its possible regulatory mechanism. Methods The protein expression of PLA2G4C in DLBCL tissues and cells was detected by Western blot. DLBCL cell lines with PLA2G4C overexpression or knockdown expression were constructed, and the cells were treated with autophagy inhibitor Chloroquine (CQ) or p38 inhibitor SB203580 for 24 h. Quantitative real time polymerase chain reaction (qRT-PCR) was used to detect the transfection efficiency of PLA2G4C; Western blot analysis was performed to detect the expression levels of PLA2G4C protein, mitochondrial autophagy related protein [microtubule-associated protein 1 light chain 3- Ⅱ / Ⅰ (LC3 Ⅱ / Ⅰ ), Beclin1, p62, PTEN induced putative kinase 1 (PINK1) and Parkin] and p38/ mitogen activated protein kinases (MAPK) pathway-related protein [Phosphorylated p38/MAPK(p-p38/MAPK)]; Cell proliferation, invasion and apoptosis were detected with CCK8, Transwell assay and the flow cytometry, respectively. The effects of PLA2G4C on tumor growth and mitochondrial autophagy in nude mice were further established. Results The PLA2G4C protein expression in lymphoma tissues of DLBCL patients was significantly higher than that in lymph nodes with reactive hyperplasia (3.47±0.42 vs 1.01±0.02)，and the difference was statistically significant(t= -37.002, P<0.001). The PLA2G4C protein level in DLBCL cells was significantly higher than that in human lymphoblast-like cell lines and B-cell lymphoma cell lines，and the difference was statistically significant (F=73.771, P<0.001). Silencing PLA2G4C significantly decreased the viability and invasion ability of DLBCL cells, and induced apoptosis, with statistical significance (t=6.909~11.390)， Overexpression of PLA2G4C gave the opposite result（t=2.392~19.778），and the differences were statistically significant(all P<0.001). PLA2G4C overexpression significantly promoted the occurrence of mitochondrial autophagy, while CQ or SB203580 treatment could significantly reverse the effects of PLA2G4C overexpression on the biological behavior of DLBCL cells and mitochondrial autophagy. In vivo nude mice experiments showed that PLA2G4C knockdown significantly inhibited tumor growth and mitochondrial autophagy related protein expression in transplanted nude mice，and the differences were statistically significant (t=13.816 ～ 25.926, all P<0.001).Conclusion PLA2G4C is up-regulated in DLBCL, which may promote tumor cell proliferation and invasion, inhibit cell apoptosis, and participate in the occurrence and development of DLBCL by promoting mitochondrial autophagy mediated by p38/MAPK signaling pathway.

参考文献/References:

[1] SMITH S M, PASQUALUCCI L. Introduction to series: diffuse large B-cell lymphoma[J]. Seminars in Hematology, 2023, 60(5): 251-254.
[2] HILTON L K, SCOTT D W, MORIN R D. Biological heterogeneity in diffuse large B-cell lymphoma[J]. Seminars in Hematology, 2023, 60(5): 267-276.
[3] LU Yingying, LI Zhijia, ZHANG Shuangqian, et al.Cellular mitophagy: mechanism, roles in diseases and small molecule pharmacological regulation[J]. Theranostics, 2023, 13(2): 736-766.
[4] NAKAHARA Y, MITSUI J, DATE H, et al. Genomewide association study identifies a new susceptibility locus in PLA2G4C for multiple system atrophy[J]. medRxiv[Preprint], 2023.doi:10.1101/2023.05.02.23289328.
[5] PENG Zhangxiao, CHANG Yanxin, FAN Jianhui, et al. Phospholipase A2 superfamily in cancer[J]. Cancer Letters, 2021, 497: 165-177.
[6] WARD K E, SENGUPTA R, ROPA J P, et al. The cytosolic phospholipase A2α N-terminal C2 domain binds and oligomerizes on membranes with positive curvature[J]. Biomolecules, 2020, 10(647): 647.
[7] OH M, JANG S Y, LEE J Y, et al. The lipoproteinassociated phospholipase A2 inhibitor Darapladib sensitises cancer cells to ferroptosis by remodelling lipid metabolism[J]. Nature Communications, 2023,14(1): 5728.
[8] OLSEN R S, ANDERSSON R E, ZAR N, et al. Prognostic s ignif icance of PLA2G4C gene polymorphism in patients with stage II colorectal cancer[J]. Acta Oncologica, 2016, 55(4): 474-479.
[9] NANASHIMA N, YAMADA T, SHIMIZU T, et al. Deletion of phospholipase A2 group IVC induces apoptosis in rat mammary tumour cells by the nuclear factor-κB/lipocalin 2 pathway[J]. Biochemical Journal, 2015, 469(2): 315-324.
[10] WANG Yunju, CHANG Songbin, WANG C Y, et al. The selective lipoprotein-associated phospholipase A2 inhibitor darapladib triggers irreversible actions on glioma cell apoptosis and mitochondrial dysfunction[J]. Toxicology and Applied Pharmacology, 2020, 402: 115133.
[11] LIM S C, LEE T B, KANG B S, et al. Extracellular acidity-mediated expression of cPLA2γ confers resistance in gastric cancer cells[J]. Anticancer Research, 2021, 41(1): 211-218.
[12] 陈思言, 张伶莉, 杨丽华.弥漫性大B 细胞淋巴瘤组织中miR-448 和KDM2B 的水平表达及临床意义[J]. 现代检验医学杂志, 2022, 37(4): 128-133. CHEN Siyan, ZHANG Lingli, YANG Lihua. Expression levels and clinical significance of miR-448 and KDM2B in diffuse large B-cell lymphoma tissues[J]. Journal of Modern Laboratory Medicine,2022, 37(4): 128-133.
[13] LI Anqi, GAO Meng, LIU Bilin, et al. Mitochondrial autophagy: molecular mechanisms and implications for cardiovascular disease[J]. Cell Death & Disease, 2022,13(5): 444.
[14] SMITH A G, MACLEOD KF. Autophagy, cancer stem cells and drug resistance[J]. Journal of Pathology, 2019,247(5): 708-718.
[15] PANIGRAHI D P, PRAHARAJ P P, BHOL C S, et al. The emerging, multifaceted role of mitophagy in cancer and cancer therapeutics[J]. Seminars in Cancer Biology,2020, 66: 45-58.
[16] 黄基峰, 张怡, 晏琛. 线粒体自噬在肿瘤干细胞中作用的研究进展[J]. 中国肿瘤临床, 2020, 47(5): 255-259. HUANG Jifeng, ZHANG Yi, YAN Chen. Research advances in the role of mitophagy in cancer stem cells[J]. Chinese Journal of Clinical Oncology, 2020,47(5): 255-259.
[17] TANG Junwei, PENG Wen, JI Jiangzhou, et al. GPR176 promotes cancer progression by interacting with G protein GNAS to restrain cell mitophagy in colorectal cancer[J]. Advanced Science, 2023, 10(12): e2205627.
[18] JIANG Ying, KRANTZ S, QIN Xiang, et al. Caveolin-1 controls mitochondrial damage and ROS production by regulating fission - fusion dynamics and mitophagy[J]. Redox Biology, 2022, 52: 102304.
[19] FENG Ji, ZHOU Jing, WU Yong, et al. Targeting mitophagy as a novel therapeutic approach in liver cancer[J]. Autophagy, 2023, 19(7): 2164-2165.
[20] LI Yun, CHEN Hengxing, LU Daning, et al. Mitophagy is a novel protective mechanism for drug-tolerant persister (DTP) cancer cells[J]. Autophagy, 2023, 19(9): 2618-2619.
[21] DENNY W A. Inhibitors and activators of the p38 mitogen-activated MAP kinase (MAPK) family as drugs to treat cancer and inflammation[J]. Current Cancer Drug Targets, 2022, 22(3): 209-220.
[22] LIU Chi, JIANG Shan, XIE Hui, et al. Long noncoding RNA AC245100.4 contributes to prostate cancer migration via regulating PAR2 and activating p38-MAPK pathway[J]. Medical Oncology, 2022, 39(5): 94.
[23] TANIGAWA K, TSUKAMOTO S, KOMA Y I, et al. S100A8/A9 induced by interaction with macrophages in esophageal squamous cell carcinoma promotes the migration and invasion of cancer cells via Akt and p38 MAPK pathways[J]. American Journal of Pathology,2022, 192(3): 536-552.
[24] KWAK A W, LEE J Y, LEE S O, et al. Echinatin induces reactive Oxygen species-mediated apoptosis via JNK/p38 MAPK signaling pathway in colorectal cancer cells[J]. Phytotherapy Research, 2023, 37(2): 563-577.

相似文献/References:

[1]陈思言,张伶莉,杨丽华.弥漫性大B 细胞淋巴瘤组织中miR-448 和KDM2B 的水平表达及临床意义[J].现代检验医学杂志,2022,37(04):128.[doi:10.3969/j.issn.1671-7414.2022.04.025]
　CHEN Si-yan,ZHANG Ling-li,YANG Li-hua.Expression Levels and Clinical Significance of miR-448 and KDM2B in Diffuse Large B-cell Lymphoma Tissues[J].Journal of Modern Laboratory Medicine,2022,37(06):128.[doi:10.3969/j.issn.1671-7414.2022.04.025]

备注/Memo

备注/Memo:
基金项目：秦皇岛市重点研发计划科技支撑项目（202101A156）。
作者简介：王敬如（1984-），女，大学本科，中级职称，研究方向：淋巴瘤，E-mail：WJR8403@163.com。
更新日期/Last Update: 2024-11-15