去泛素化酶USP35促进非小细胞肺癌细胞迁移和侵袭

doi:10.6040/j.issn.1671-7554.0.2021.1108

摘要/Abstract

摘要： 目的探究泛素特异性蛋白酶35(USP35)对非小细胞肺癌(NSCLC)侵袭、迁移能力的影响。方法 Western blotting检测7种NSCLC细胞中USP35蛋白的表达。根据Lipofectamine 2000说明书,USP35过表达或对照质粒转入H1299细胞,干扰或对照质粒转入Anip973细胞。Transwell无基底胶小室检测细胞迁移能力,预加入matrigel基底胶包被小室检测细胞侵袭能力。Western blotting检测USP35蛋白过表达和干扰时相应的上皮钙黏蛋白(Ecadherin)、波形蛋白(Vimentin)表达水平。结果选取的7种NSCLC细胞中,H1299细胞USP35蛋白表达最低、Anip973细胞USP35蛋白表达最高。与转染对照质粒相比,转染过表达USP35质粒后的H1299细胞迁移和侵袭的数目明显增加,E-cadherin表达下调,Vimentin表达上调。与转染对照质粒相比,转染USP35干扰质粒后的Anip973细胞迁移和侵袭的数目明显降低,E-cadherin表达上调,Vimentin表达下调。结论 USP35在NSCLC细胞迁移和侵袭中发挥重要作用,USP35可能成为NSCLC潜在的治疗选择。

关键词: 泛素特异性蛋白酶35, 非小细胞肺癌, 侵袭, 迁移, 上皮间质转化

Abstract: Objective To investigate the effects of deubiquitinase ubiquitin-speci cproteases 35(USP35)on the migration and invasion of non-small cell lung cancer(NSCLC). Methods The protein expression of USP35 in a panel of 7 human NSCLC cell lines was examined with Western blotting. USP35 overexpression plasmids and control plasmids were transfected into H1299 cells, while USP35 knockdown plasmids and control plasmids were transfected into Anip973 cells according to Lipofectamine 2000 instructions. The migration of NSCLC cells was analyzed using Transwell blank chambers, and the invasion was analyzed using the Transwell chambers containing matrigel substrate gels. The protein expressions of E-cadherin and Vimentin were detected with Western blotting after USP35 overexpressing or silencing in aforementioned cell lines. Results Of the 7 NSCLC cell lines tested, USP35 expression was the lowest in H1299 cells, and the highest in Anip973 cells. Compared with transfected USP35 control plasmids, transfected USP35 overexpression plasmids had significantly increased migration and invasion, down-regulated E-cadherin, and up-regulated Vimentin expressions. Compared with transfected USP35 control plasmids, the Anip973 cells of transfected USP35 knockdown plasmids had significantly reduced migration and invasion, up-regulated E-cadherin, and down-regulated Vimentin expressions. Conclusion USP35 plays a critical role in the migration and invasion of NSCLC cells, hence may become a therapeutic option for NSCLC.

Key words: Ubiquitin-speciı, c proteases 35, Non-small cell lung cancer, Invasion, Migration, Epithelial-mesenchymal transition

中图分类号:

R574

陈兆波,方敏,薛浩然,刘春艳. 去泛素化酶USP35促进非小细胞肺癌细胞迁移和侵袭[J]. 山东大学学报 (医学版), 2022, 60(4): 30-37.

CHEN Zhaobo, FANG Min, XUE Haoran, LIU Chunyan. Deubiquitinase USP35 promotes the cells migration and invasion of non-small cell lung cancer[J]. Journal of Shandong University (Health Sciences), 2022, 60(4): 30-37.

参考文献

[1] Chen W, Zheng R, Baade PD, et al. Cancer statistics in China, 2015 [J]. CA Cancer J Clin, 2016, 66(2): 115-132.
[2] Siegel RL, Miller KD, Fuchs HE, et al. Cancer Statistics, 2021 [J]. CA Cancer J Clin, 2021, 71(1): 7-33.
[3] Samson P, Keogan K, Crabtree T, et al. Interpreting survival data from clinical trials of surgery versus stereotactic body radiation therapy in operable Stage I non-small cell lung cancer patients [J]. Lung Cancer, 2017, 103: 6-10. doi: 10.1016/j.lungcan.2016.11.005.
[4] Beckett P, Tata LJ, Hubbard RB. Risk factors and survival outcome for non-elective referral in non-small cell lung cancer patients - analysis based on the National Lung Cancer Audit [J]. Lung cancer, 2014, 83(3): 396-400.
[5] Miller KD, Nogueira L, Mariotto AB, et al. Cancer treatment and survivorship statistics, 2019 [J]. CA Cancer J Clin, 2019, 69(5): 363-385.
[6] Xiao Y, Yin C, Wang Y, et al. FBXW7 deletion contributes to lung tumor development and confers resistance to gefitinib therapy [J]. Mol Oncol, 2018, 12(6): 883-895.
[7] 简政, 陈学瑜, 张亚杰, 等. 2022 年 V1 版《NCCN 非小细胞肺癌临床实践指南》更新解读[J]. 中国胸心血管外科临床杂志, 2022, 29(2): 150-157. JIAN Zheng, CHEN Xueyu, ZHANG Yajie, et al. Interpretation of updated NCCN guidelines for non-small cell lung cancer(version 1, 2022)[J]. Chinese Journal of Clinical Thoracic and Cardiovascular Surgery, 2022, 29(2): 150-157.
[8] 周广华, 林琳. 靶向治疗在 NSCLC 中的研究进展[J]. 临床肺科杂志, 2020, 25(4): 602-605.
[9] 戴陈新, 袁健, 郑亚新, 等. 去泛素化酶与肿瘤发生及发展关系的研究进展[J]. 中国癌症杂志, 2013, 23(7): 547-550. DAI Chenxin, YUAN Jian, ZHENG Yaxin, et al. Advances in the study of deubiquitinating enzymes in cancer research [J]. China Oncology, 2013, 23(7): 547-550.
[10] Wang W, Wang M, Xiao Y, et al. USP35 mitigates endoplasmic reticulum stress-induced apoptosis by stabilizing RRBP1 in non-small cell lung cancer [J]. Mol Oncol, 2021. doi: 10.1002/1878-0261.13112.
[11] Park J, Kwon MS, Kim EE, et al. USP35 regulates mitotic progression by modulating the stability of Aurora B [J]. Nat Commun, 2018, 9(1): 688.
[12] Cao J, Wu D, Wu G, et al. USP35, regulated by estrogen and AKT, promotes breast tumorigenesis by stabilizing and enhancing transcriptional activity of estrogen receptor α [J]. Cell Death Dis, 2021, 12(6): 619.
[13] Clague MJ, Urbé S, Komander D. Breaking the chains: deubiquitylating enzyme specificity begets function [J]. Nat Rev Mol Cell Biol, 2019, 20(6): 338-352.
[14] Harrigan JA, Jacq X, Martin NM, et al. Deubiquitylating enzymes and drug discovery: emerging opportunities [J]. Nat Rev Drug Discov, 2018, 17(1): 57-78.
[15] Liu S, González-Prieto R, Zhang M, et al. Deubiquitinase activity profiling identifies UCHL1 as a candidate oncoprotein that promotes TGFβ-induced breast cancer metastasis [J]. Clin Cancer Res, 2020, 26(6): 1460-1473.
[16] Zhang HH, Li C, Ren JW, et al. OTUB1 facilitates bladder cancer progression by stabilizing ATF6 in response to endoplasmic reticulum stress [J]. Cancer Sci, 2021, 112(6): 2199-2209.
[17] Wu X, Luo Q, Zhao P, et al. JOSD1 inhibits mitochondrial apoptotic signalling to drive acquired chemoresistance in gynaecological cancer by stabilizing MCL1 [J]. Cell Death Differ, 2020, 27(1): 55-70.
[18] Khan OM, Carvalho J, Spencer-Dene B, et al. The deubiquitinase USP9X regulates FBW7 stability and suppresses colorectal cancer [J]. J Clin Invest, 2018, 128(4): 1326-1337.
[19] Tanguturi P, Kim KS, Ramakrishna S. The role of deubiquitinating enzymes in cancer drug resistance [J]. Cancer Chemother Pharmacol, 2020, 85(4): 627-639.
[20] Liao XH, Wang Y, Zhong B, et al. USP3 promotes proliferation of non-small cell lung cancer through regulating RBM4 [J]. Eur Rev Med Pharmacol Sci, 2020, 24(6): 3143-3151.
[21] Wu Y, Qin J, Li F, et al. USP3 promotes breast cancer cell proliferation by deubiquitinating KLF5 [J]. J Biol Chem, 2019, 294(47): 17837-17847.
[22] Garcia DA, Baek C, Estrada MV, et al. USP11 enhances TGFβ-induced epithelial-mesenchymal plasticity and human breast cancer metastasis [J]. Mol Cancer Res, 2018, 16(7): 1172-1184.
[23] Sun H, Ou B, Zhao S, et al. USP11 promotes growth and metastasis of colorectal cancer via PPP1CA-mediated activation of ERK/MAPK signaling pathway [J]. EbioMedicine, 2019, 48: 236-247. doi: 10.1016/j.ebiom.2019.08.061.
[24] Zhang C, Xie C, Wang X, et al. Aberrant USP11 expression regulates NF90 to promote proliferation and metastasis in hepatocellular carcinoma [J]. Am J Cancer Res, 2020, 10(5): 1416-1428.
[25] Ha J, Kim M, Seo D, et al. The deubiquitinating enzyme USP20 regulates the TNFα-induced NF-κB signaling pathway through stabilization of p62 [J]. Int J Mol Sci, 2020, 21(9): 3116.
[26] Li W, Shen M, Jiang YZ, et al. Deubiquitinase USP20 promotes breast cancer metastasis by stabilizing SNAI2 [J]. Genes Dev, 2020, 34(19-20): 1310-1315.
[27] Wu C, Luo K, Zhao F, et al. USP20 positively regulates tumorigenesis and chemoresistance through β-catenin stabilization [J]. Cell Death Differ, 2018, 25(10): 1855-1869.
[28] Gan Q, Shao J, Cao Y, et al. USP33 regulates c-Met expression by deubiquitinating SP1 to facilitate metastasis in hepatocellular carcinoma [J]. Life Sci, 2020, 261:118316. doi: 10.1016/j.lfs.2020.118316.
[29] Han PP, Zhang GQ, Li L, et al. Downregulation of USP33 inhibits Slit/Robo signaling pathway and is associated with poor patient survival of glioma [J]. J Neurosurg Sci, 2020. doi: 10.23736/S0390-5616.20.04929-2.
[30] Huang Z, Wen P, Kong R, et al. USP33 mediates Slit-Robo signaling in inhibiting colorectal cancer cell migration [J]. Int J Cancer, 2015, 136(8): 1792-1802.
[31] Xia Y, Wang L, Xu Z, et al. Reduced USP33 expression in gastric cancer decreases inhibitory effects of Slit2-Robo1 signalling on cell migration and EMT [J]. Cell Prolif, 2019, 52(3): e12606.
[32] Dou N, Hu Q, Li L, et al. USP32 promotes tumorigenesis and chemoresistance in gastric carcinoma via upregulation of SMAD2 [J]. Int J Biol Sci, 2020, 16(9): 1648-1657.
[33] Hu W, Wei H, Li K, et al. Downregulation of USP32 inhibits cell proliferation, migration and invasion in human small cell lung cancer [J]. Cell Prolif, 2017, 50(4): e12343.
[34] Liu C, Wang L, Chen W, et al. USP35 activated by miR let-7a inhibits cell proliferation and NF-κB activation through stabilization of ABIN-2 [J]. Oncotarget, 2015, 6(29): 27891-27906.
[35] Wang Y, Serricchio M, Jauregui M, et al. Deubiquitinating enzymes regulate PARK2-mediated mitophagy [J]. Autophagy, 2015, 11(4): 595-606.
[36] Zhang J, Chen Y, Chen X, et al. Deubiquitinase USP35 restrains STING-mediated interferon signaling in ovarian cancer [J]. Cell Death Differ, 2021, 28(1): 139-155.
[37] Tang Z, Jiang W, Mao M, et al. Deubiquitinase USP35 modulates ferroptosis in lung cancer via targeting ferroportin [J]. Clin Transl Med, 2021, 11(4): e390.

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