黑色素瘤SENP1蛋白质参与达卡巴嗪耐药性的探究

doi:10. 3969/ j. issn. 1006-7795. 2024. 01. 016

首都医科大学学报 ›› 2024, Vol. 45 ›› Issue (1): 97-103.doi: 10. 3969/ j. issn. 1006-7795. 2024. 01. 016

黑色素瘤SENP1蛋白质参与达卡巴嗪耐药性的探究

赵蓓¹，施小琪²，唐雪梅¹，程石^1*

1.四川省医学科学院四川省人民医院(电子科技大学附属医院)皮肤科（皮肤病性病研究所），成都 610000；2.成都中医药大学医学与生命科学学院，成都 610000

收稿日期:2023-09-11 出版日期:2024-02-21 发布日期:2024-03-22
通讯作者: 程石 E-mail:ruiruishisan@126.com
基金资助:
四川省自然科学基金青年项目（2023NSFSC1549），四川省干部保健科研项目(川干研2023-218)，四川省医学青年创新科研课题计划（Q21043）。

Involvement of SENP1 protein in resistance of dacarbazine in melanoma

Zhao Bei¹，Shi Xiaoqi²，Tang Xuemei¹，Cheng Shi^1*

1.Department of Dermatology (Institute of Dermatology and Venereology),Sichuan Academy of Medical Sciences and Sichuan Provincial People's Hospital,University of Electronic Science and Technology of China,Chengdu 610000， China； 2.School of Medical and Life Sciences,Chengdu University of Traditional Chinese Medicine,Chengdu 610000,China

Received:2023-09-11 Online:2024-02-21 Published:2024-03-22
Supported by:
This study was supported by the Youth Program of Natural Science Foundation of Sichuan Province (2023NSFSC1549),The Cadre Health Care Research Project of Sichuan Province (2023-218), Youth Innovation Project of Sichuan Medical Association (Q21043).

摘要/Abstract

摘要： 目的探究与黑色素瘤耐药性相关的基因及信号通路，揭示其与黑色素瘤耐药性的相关性。方法以A375及M14黑色素瘤细胞为研究对象，通过逐渐提高达卡巴嗪（dacarbazine，DTIC）的浓度获得耐药性黑色素瘤细胞株，采用转录物组学研究耐药性黑色素瘤细胞系中显著性变化的基因及通路，利用实时荧光定量聚合酶链反应（real time quantitative polymerase chain reaction，RT-qPCR）、蛋白质杂交（Western blotting，WB）等对变化的基因进行验证。结果 （1）成功构建了黑色素瘤耐药细胞株：通过DTIC小剂量逐步增加的方法成功建立了耐药型的黑色素瘤细胞系A375与M14，通过计算其对DTIC的半抑制浓度值（the half maximal inhibitory concentration，IC50），确定了细胞对DTIC的敏感性发生了显著的变化；通过流式细胞技术检测，发现耐药的黑色素瘤细胞具有显著抗DTIC引发的凋亡的能力。（2）发现了黑色素瘤耐药性相关的基因及信号通路：利用建立的耐DTIC的黑色素瘤细胞系，进行了全基因组转录测序和分析，发现了类泛素特异性蛋白酶1（SUMO-specific protease 1，SENP1）的高表达和蛋白激酶Hippo信号通路的异常活化相关。（3）SENP1异常表达可能参与DTIC耐药：WB检测野生型和耐药型黑色素瘤细胞系发现在耐药的细胞中，SENP1与YAP表达都上调。（4）通过基因敲除证实SENP1参与DTIC耐药，蛋白质相互作用实验初步证实SENP1对YAP存在去泛素化调控作用。结论 SENP1与DTIC耐药性存在正相关关系，其异常上调可能导致Hippo信号通路发生变化使得黑色素瘤对DTIC耐受性的提升。

关键词: 黑色素瘤, 耐药性, 转录物组学, 类泛素特异性蛋白酶, Hippo信号通路

Abstract: Objective To investigate the genes and signaling pathways associated with drug resistance in melanoma and establish their relationship with melanoma drug resistance. Methods A375 and M14 melanoma cells were used as the experimental model to develop a drug-resistant melanoma cell line by gradually increasing the concentration of dacarbazine (DTIC). Transcriptomics was employed to analyze significant changes in genes and pathways within the drug-resistant melanoma cell line. Real-time quantitative polymerase chain reaction (RT-qPCR) and Western blotting(WB) were conducted to validate the altered genes. Results (1) Successful construction of melanoma drug-resistant cell lines: drug-resistant cell lines,namely A375 and M14,were successfully established by gradually increasing the dosage of DTIC. The determination of their the half maximal inhibitory concentration (IC50) values to DTIC indicated a significant change in cell sensitivity. Flow cytometry analysis revealed that drug-resistant melanoma cells exhibited a notable ability to resist apoptosis induced by DTIC. (2) Identification of genes and signaling pathways associated with melanoma drug resistance: transcriptomic analysis by the established DTIC-resistant melanoma cell lines demonstrated that increased expression of SENP1 was correlated with abnormal activation of the Hippo signaling pathway. (3) Implication of aberrant expression of ubiquitin-specific protease SENP1 in DTIC resistance: WB analysis of wild-type and drug-resistant melanoma cell lines revealed upregulation of both SENP1 and YAP expressions in drug-resistant cells. (4) The involvement of SENP1 in DTIC resistance was confirmed by gene knockout and the effect of SENP1 on ubiquitination of YAP was preliminarily confirmed by protein interaction experiments. Conclusion A positive correlation existed between SENP1 and DTIC resistance in melanoma,which may contribute to alterations in the Hippo signaling pathway and enhance melanoma tolerance to DTIC.

Key words: melanoma, drug resistance, transcriptome, Ubiquitin-like specific proteases, Hippo signaling pathway

中图分类号:

R751

赵蓓, 施小琪, 唐雪梅, 程石. 黑色素瘤SENP1蛋白质参与达卡巴嗪耐药性的探究[J]. 首都医科大学学报, 2024, 45(1): 97-103.

Zhao Bei, Shi Xiaoqi, Tang Xuemei, Cheng Shi. Involvement of SENP1 protein in resistance of dacarbazine in melanoma[J]. Journal of Capital Medical University, 2024, 45(1): 97-103.

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链接本文: https://journal03.magtech.org.cn/Jweb_sdykdxxb/CN/10. 3969/ j. issn. 1006-7795. 2024. 01. 016

https://journal03.magtech.org.cn/Jweb_sdykdxxb/CN/Y2024/V45/I1/97

参考文献

［1］Kalal B S, Upadhya D, Pai V R. Chemotherapy resistance mechanisms in advanced skin cancer［J］. Oncol Rev, 2017, 11(1): 326.
［2］Sung H, Ferlay J, Siegel R L, et al. Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries［J］. CA Cancer J Clin, 2021, 71(3): 209-249.
［3］Wu S, Singh R K. Resistance to chemotherapy and molecularly targeted therapies: rationale for combination therapy in malignant melanoma［J］. Curr Mol Med, 2011, 11(7): 553-563.
［4］代强, 孙充洲, 王帅道, 等. 皮肤恶性黑色素瘤的治疗进展［J］. 医学综述, 2020, 26(15): 2982-2985, 2991.
［5］罗毅, 陈佳, 李先安, 等. 重组人血管内皮抑制素(恩度)联合达卡巴嗪治疗晚期黑色素瘤的临床研究［J］. 肿瘤药学, 2017, 7(3): 341-344.
［6］孙金中, 孙圣荣, 蒋冠. 达卡巴嗪诱导黑素瘤耐药细胞系的建立及其耐药性［J］. 临床与病理杂志, 2017, 37(11): 2305-2309.
［7］梁亚奇, 段彤. SENP 1与恶性肿瘤相关性的研究进展［J］. 重庆医学, 2023, 52(5): 776-781.
［8］Hu H J, Ling B, Shi Y H, et al. Plasma exosome-derived SENP1 may be a potential prognostic predictor for melanoma［J］. Front Oncol, 2021, 11: 685009.
［9］王明珠, 周阳, 何凯桐, 等. Hippo通路在肿瘤发生发展中作用机制的研究进展［J］. 现代肿瘤医学, 2022, 30(16): 3061-3064.
［10］Vittoria M A, Kingston N, Kotynkova K, et al. Inactivation of the hippo tumor suppressor pathway promotes melanoma［J］. Nat Commun, 2022, 13(1): 3732.
［11］Zhao B, Xie J, Zhou X Y, et al. YAP activation in melanoma contributes to anoikis resistance and metastasis［J］. Exp Biol Med (Maywood), 2021, 246(8): 888-896.
［12］Pópulo H, Soares P, Lopes J M. Insights into melanoma: targeting the mTOR pathway for therapeutics［J］. Expert Opin Ther Targets, 2012, 16(7): 689-705.
［13］Domingues B, Lopes J M, Soares P, et al. Melanoma treatment in review［J］. Immunotargets Ther, 2018, 7: 35-49.
［14］Siegel R, DeSantis C, Virgo K, et al. Cancer treatment and survivorship statistics, 2012［J］. CA Cancer J Clin, 2012, 62(4): 220-241.
［15］Li J, Wang Y, Liang R, et al. Recent advances in targeted nanoparticles drug delivery to melanoma［J］. Nanomedicine, 2015, 11(3): 769-794.
［16］Austin E, Mamalis A, Ho D, et al. Laser and light-based therapy for cutaneous and soft-tissue metastases of malignant melanoma: a systematic review［J］. Arch Dermatol Res, 2017, 309(4): 229-242.
［17］Jia Y Y, Guo Y T, Jin Q, et al. A SUMOylation-dependent HIF-1α/CLDN6 negative feedback mitigates hypoxia-induced breast cancer metastasis［J］. J Exp Clin Cancer Res, 2020, 39(1): 42.
［18］Wang X W, Liang X J, Liang H, et al. SENP1/HIF-1α feedback loop modulates hypoxia-induced cell proliferation, invasion, and EMT in human osteosarcoma cells［J］. J Cell Biochem, 2018, 119(2): 1819-1826.
［19］Zhu S B, Hu J H, Cui Y H, et al. Knockdown of SENP1 inhibits HIF-1α SUMOylation and suppresses oncogenic CCNE1 in Wilms tumor［J］. Mol Ther Oncolytics, 2021, 23: 355-366.
［20］Song Z, Wang J F, Su Q H, et al. The role of MMP-2 and MMP-9 in the metastasis and development of hypopharyngeal carcinoma［J］. Braz J Otorhinolaryngol, 2021, 87(5): 521-528.
［21］Gao C C, Xiao F J, Zhang L, et al. SENP1 inhibition suppresses the growth of lung cancer cells through activation of A20-mediated ferroptosis［J］. Ann Transl Med, 2022, 10(4): 224.
［22］Chen M C, Nhan D C, Hsu C H, et al. SENP1 participates in Irinotecan resistance in human colon cancer cells［J］. J Cell Biochem, 2021, 122(10): 1277-1294.

黑色素瘤SENP1蛋白质参与达卡巴嗪耐药性的探究

Involvement of SENP1 protein in resistance of dacarbazine in melanoma

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