p62蛋白在食管鳞癌细胞中核浆易位的机制研究

doi:10.3969/j.issn.1004-616x.2021.05.004

Abstract

Abstract: OBJECTIVE: To explore possible mechanisms that mediated the translocation of p62 protein from nuclei into cytoplasm in esophageal squamous cell carcinoma. METHODS: After treatments with KPT330 in KYSE70,KYSE150 and KYSE510 ESCC cell lines,localizations of p62 were observed by immunofluorescence assay combined with confocal laser microscopy as compared with the untreated group. LC-MS/MS mass spectrometry was used to identify the phosphorylation sites of p62 in esophageal cancer cells. Site-directed mutagenesis was performed to construct mutants with different phosphorylation status of p62. Co-IP and PLA experiments were conducted to analyze interactions between p62 and GSK3β. p62 accumulated in the nucleus after treatment with nuclear export inhibitor. RESULTS: The results of mass spectrometry showed phosphorylation of p62 protein at four sites of T269/S272/S328/S332 in esophageal cancer cells. By transfecting different phosphorylated mutants of p62 into KYSE30 cells and KYSE150 cells with endogenous knockdown of p62,phosphorylation at T269/S272 and dephosphorylation of S328/S332 contributed to the transfer of p62 from cytoplasm to nucleus. Co-IP and PLA results showed direct interactions between p62 and GSK-3β. CONCLUSION: Our results indicates that nucleo-plasmic translocation of p62 in esophageal cancer cells was related to the phosphorylation status of S328/S332 and might be regulated by GSK-3β kinase.

Key words: p62 protein, esophageal squamous cell carcinoma, nucleo-plasma translocation, phosphorylation, GSK-3β

CLC Number:

R73.02

YANG Liyan, LIU Zou, HAO Jiajie, ZHANG Yu, WANG Mingrong. Study on the mechanism of nucleo-plasma translocation of p62 protein in esophageal squamous cell carcinoma[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2021, 33(5): 338-344.

References

[1] CHEN W, ZHENG R, BAADE P D, et al. Cancer statistics in China, 2015[J]. CA:Cancer J Clin, 2016, 66(2):115-132.
[2] LAGERGREN J, SMYTH E, CUNNINGHAM D, et al. Oesophageal cancer[J]. Lancet, 2017, 390(10110):2383-2396.
[3] PENNATHUR A, GIBSON M K, JOBE B A, et al. Oesophageal carcinoma[J]. Lancet, 2013, 381(9864):400-412.
[4] SHI C, PAN B Q, SHI F, et al. Sequestosome 1 protects esophageal squamous carcinoma cells from apoptosis via stabilizing SKP2 under serum starvation condition[J]. Oncogene, 2018, 37(24):3260-3274.
[5] SÁNCHEZ-MARTÍN P, SAITO T, KOMATSU M. p62/SQSTM1:'Jack of all trades'in health and cancer[J]. Febs J, 2019, 286(1):8-23.
[6] LI D, HE C, YE F, et al. p62 overexpression promotes bone metastasis of lung adenocarcinoma out of LC3-dependent autophagy[J]. Front Oncol, 2021, 11:609548.
[7] YOKOTA A, HIRAMOTO M, HINO H, et al. Sequestosome 1(p62) accumulation in breast cancer cells suppresses progesterone receptor expression via argonaute 2[J]. Biochem Biophys Res Commun, 2020, 531(2):256-263.
[8] MAO Y, DENG S J, SU Y J, et al. The role of P62 in the development of human thyroid cancer and its possible mechanism[J]. Cancer Genet, 2021, 256/257:5-16.
[9] LEI C, ZHAO B, LIU L, et al. Expression and clinical significance of p62 protein in colon cancer[J]. Medicine:Baltimore, 2020, 99(3):e18791.
[10] BURDELSKI C, REISWICH V, HUBE-MAGG C, et al. Cytoplasmic accumulation of sequestosome 1(p62) is a predictor of biochemical recurrence, rapid tumor cell proliferation, and genomic instability in prostate cancer[J]. Clin Cancer Res, 2015, 21(15):3471-3479.
[11] PANKIV S, LAMARK T, BRUUN J A, et al. Nucleocytoplasmic shuttling of p62/SQSTM1 and its role in recruitment of nuclear polyubiquitinated proteins to promyelocytic leukemia bodies[J]. J Biol Chem, 2010, 285(8):5941-5953.
[12] ISLAM M, SOORO M, ZHANG P H. Autophagic regulation of p62 is critical for cancer therapy[J]. Int J Mol Sci, 2018, 19(5):1405.
[13] WANG S B, VENKATRAMAN V, CROWGEY E L, et al. Protein S-nitrosylation controls glycogen synthase kinase 3β function independent of its phosphorylation state[J]. Circ Res, 2018, 122(11):1517-1531.
[14] FRAME S, COHEN P. GSK₃ takes centre stage more than 20 years after its discovery[J]. Biochem J, 2001, 359(pt 1):1-16.
[15] JOPE R S. Lithium and GSK-3:one inhibitor, two inhibitory actions, multiple outcomes[J]. Trends Pharmacol Sci, 2003, 24(9):441-443.
[16] JOPE R S, JOHNSON G V. The glamour and gloom of glycogen synthase kinase-3[J]. Trends Biochem Sci, 2004, 29(2):95-102.
[17] LIN J T, SONG T, LI C, et al. GSK-3β in DNA repair, apoptosis, and resistance of chemotherapy, radiotherapy of cancer[J]. Biochim et Biophys Acta BBA Mol Cell Res, 2020, 1867(5):118659.
[18] GUO L, CHEN D K, YIN X, et al. GSK-3β promotes cell migration and inhibits autophagy by mediating the AMPK pathway in breast cancer[J]. Oncol Res, 2019, 27(4):487-494.
[19] CHEN R, ZHANG Y Y, LAN J N, et al. Ischemic postconditioning alleviates intestinal ischemia-reperfusion injury by enhancing autophagy and suppressing oxidative stress through the Akt/GSK-3β/Nrf2 pathway in mice[J]. Oxid Med Cell Longev, 2020, 2020:6954764.
[20] REN J L, LIU T T, HAN Y, et al. GSK-3β inhibits autophagy and enhances radiosensitivity in non-small cell lung cancer[J]. Diagn Pathol, 2018, 13(1):1-10.
[21] WEIBRECHT I, LEUCHOWIUS K J, CLAUSSON C M, et al. Proximity ligation assays:a recent addition to the proteomics toolbox[J]. Expert Rev Proteom, 2010, 7(3):401-409.

Study on the mechanism of nucleo-plasma translocation of p62 protein in esophageal squamous cell carcinoma

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	YANG Xingxiao, ZHANG Xueyuan, ZOU Naiyi, SHAN Bao'en, MA Ming, ZHU Shuchai. Effects of HMGB1 silencing on migration, invasion and radiosensitivity of esophageal squamous cell carcinoma cells [J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2021, 33(5): 327-333.
[2]	CHANG Chen, YANG Liyan, CAI Hongqing, YUAN Qing, HAO Jiajie, WANG Mingrong. Expressions and clinical implications of SERPINE1 mRNA in esophageal squamous carcinoma tissues [J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2021, 33(4): 245-249.
[3]	ZENG Zhuoying, WU Desheng, LU Jingjing, LIAO Hui, LAI Hongpiao, YUAN Jianhui, HU Zhangli. Mediation by estrogen receptor α on oxidative phosphorylation pathways in development of lung cancer [J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2021, 33(4): 262-268.
[4]	ZHAO Li, WANG Li, MO Ling, SUN Yan. Effects of liraglutide on cognitive function and phosphorylation of Tau protein in streptozotocin-induced Alzheimer in rats [J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2021, 33(4): 280-285.
[5]	ZHENG Junwen, CHANG Chen, HAO Jiajie, CAI Yan, WANG Mingrong, ZHANG Yu. Investigation on expression and function of NEDD4 in esophageal squamous cell carcinoma [J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2021, 33(3): 203-207,224.
[6]	JIAO Wenjing, GUO Xiujuan, SHAO Junguo, JIAO Wenpeng, ZHANG Jinyan. Relationships among blood cell ratios, clinicopathological characteristics and prognosis in esophageal squamous cell carcinoma [J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2021, 33(3): 213-217.
[7]	ZHANG Die, LIU Xianghe, HUO Miaomiao, LIU Mei, XU Ningzhi, ZHU Hongxia. Effects of miR-195-5p on radiosensitivity of esophageal squamous cell carcinoma cells [J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2021, 33(2): 81-88,94.
[8]	ZHANG Na, FAN Zhilu, YANG Liyan, CHANG Chen, CAI Yan, HAO Jiajie, WANG Mingrong. Clinical significance of decreased SERPINB5 expression in esophageal squamous cell carcinomas [J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2021, 33(2): 101-109.
[9]	GULZARIA·Aikula, TAN Yao, PALIDA·Apiziaji. Expression of long non-coding RNAs in esophageal squamous cell carcinoma of Kazak nationality in Xinjiang [J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2020, 32(3): 194-197,202.
[10]	SHI Pan, LIN Chonghua, SHU Yifang, QU Jiaquan, SONG Shuliang. Role of mitochondrial citrate transporter SLC25A1 in radiosensitivity of human esophageal squamous cell carcinoma in vitro [J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2020, 32(2): 132-138.
[11]	SUN Xiaonan, YUAN Qing, YANG Liyan, ZHANG Yu, CAI Yan, XU Xin, WANG Mingrong, HAO Jiajie, JIA Xuemei. FGFBP2 over-expresssion promoted cell proliferation in esophageal squamous carcinoma cells [J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2019, 31(3): 180-185,192.
[12]	HUANG Pengcheng, ZHOU Changhui, LI Shenning, CHANG Yan. Establishment of detection method for histone γ-H2AX phosphorylation based on flow cytometry [J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2017, 29(4): 284-288.
[13]	XIE Zhihui, YANG Hai, ZHANG Yu, HAO Jiajie, CAI Yan, XU Xin, WANG Mingrong. Analysis of expression of KIAA1522 in esophageal squamous cell carcinoma [J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2017, 29(3): 166-171.
[14]	GAO Hua, SONG Tieniu, LI Bin, YANG Jianbao, ZHU Duojie, GUO Quanwei. Effect of RNA helicase gene DDX46 knockdown on proliferation and apoptosis in esophageal squamous cell carcinoma Eca-109 cells [J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2017, 29(3): 216-221,229.
[15]	CHANG Chen, ZHAO Xiaohan, JIANG Ye, XU Xin, CAI Yan, PEI Yuhui, ZHANG Yu, HAO Jiajie, WANG Mingrong. Identification of predominant genome DNA alterations and their clinical implication in esophageal squamous cell carcinoma [J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2017, 29(1): 1-6.