您的位置：山东大学 -> 科技期刊社 -> 《山东大学学报（医学版）》

山东大学学报 (医学版) ›› 2023, Vol. 61 ›› Issue (1): 17-26.doi: 10.6040/j.issn.1671-7554.0.2022.1064

• 基础医学 • 上一篇    下一篇

长链非编码RNA ZNF528-AS1促进乳腺癌他莫昔芬耐药及进展转移

张建树1,张瀚文1,赵文静2   

  1. 山东大学齐鲁医院 1. 乳腺外科;2.生物样本资源库, 山东 济南 250012
  • 发布日期:2023-01-10
  • 通讯作者: 赵文静. E-mail:zhaowenjing0911@163.com
  • 基金资助:
    中国博士后科学基金(2017M612292)

LncRNA ZNF528-AS1 promotes tamoxifen resistance and progression of breast cancer

ZHANG Jianshu1, ZHANG Hanwen1, ZHAO Wenjing2   

  1. 1. Department of Breast Surgery;
    2. Pathology Tissue Bank, Qilu Hospital of Shandong University, Jinan 250012, Shandong, China
  • Published:2023-01-10

PDF (PC)

283

摘要： 目的 探讨长链非编码RNA ZNF528-AS1对乳腺癌他莫昔芬耐药及进展转移的影响及潜在调控机制。 方法 利用RNA-seq高通量测序技术分析乳腺癌他莫昔芬耐药细胞/成瘤组织与亲本细胞/成瘤组织间差异表达的lncRNA,筛选与乳腺癌他莫西芬耐药相关的候选lncRNA,利用实时定量PCR(qRT-PCR)技术验证候选lncRNA的表达。通过脂质体转染乳腺癌细胞,构建过表达及敲低ZNF528-AS1的细胞模型。利用细胞增殖实验、克隆形成实验、成球实验、5-乙炔基-2'-脱氧尿嘧啶核苷(EdU)实验检测ZNF528-AS1对乳腺癌细胞增殖、肿瘤干性及他莫昔芬耐药性的影响。Transwell迁移和侵袭实验检测ZNF528-AS1对乳腺癌细胞转移能力的影响。利用RNA-seq高通量测序及生物信息学分析技术寻找ZNF528-AS1的下游调控通路,并通过Western blotting及细胞功能实验等进行验证。 结果 RNA-seq高通量测序分析筛选出了在他莫昔芬耐药细胞/成瘤组织与亲本细胞/成瘤组织间差异表达的lncRNAs。qRT-PCR实验结果证实了差异lncRNA的表达,并发现ZNF528-AS1在他莫昔芬耐药细胞中的表达上调最为明显。在乳腺癌细胞中过表达ZNF528-AS1能够提高细胞的增殖水平、克隆形成能力、肿瘤干性及对他莫昔芬的耐药性。此外,过表达ZNF528-AS1可增强乳腺癌细胞的迁移及侵袭能力。相应地,在乳腺癌细胞中敲低ZNF528-AS1能够降低细胞的增殖、他莫昔芬耐药性、肿瘤干性、迁移及侵袭等能力。RNA-seq高通量测序结合生物信息学分析结果显示,过表达ZNF528-AS1能够参与调控转化生长因子-β(TGF-β)信号通路。Western blotting结果证实ZNF528-AS1能够同时激活经典和非经典TGF-β信号通路。TGF-β通路抑制剂SB431542能够逆转ZNF528-AS1过表达增强的细胞增殖、迁移及侵袭能力。 结论 ZNF528-AS1能够促进乳腺癌他莫昔芬耐药及进展转移,其作用机制可能与激活TGF-β通路有关。

关键词: 乳腺癌, ZNF528-AS1, 他莫昔芬耐药, 肿瘤干性, 进展转移

Abstract: Objective To investigate the effects of long non-coding RNA(LncRNA)ZNF528-AS1 on tamoxifen resistance and progression of breast cancer as well as the potential regulatory mechanism. Methods The differentially expressed lncRNAs among tamoxifen-resistant breast cancer cells/xenograft tissues and parent cells/ xenograft tissues were analyzed with RNA-seq high-throughput sequencing technology, and candidate lncRNAs associated with tamoxifen resistance were screened. The expressions of candidate lncRNAs were verified with quantitative real-time PCR(qRT-PCR). Breast cancer cells were transfected with liposomes to construct ZNF528-AS1 overexpression and knockdown cell models. The effects of ZNF528-AS1 on the proliferation, tumor stemness and tamoxifen resistance were detected with MTT, colony formation, mammosphere formation, and EdU assays. The role of ZNF528-AS1 in the metastasis of breast cancer was evaluated with Transwell migration and invasion assays. The downstream regulatory pathways of ZNF528-AS1 were detected with RNA-seq high-throughput sequencing and bioinformatics analysis, and verified with Western blotting and functional experiments. Results The differentially expressed lncRNAs were screened out and verified. ZNF528-AS1 was significantly upregulated in tamoxifen-resistant cells. Overexpression of ZNF528-AS1 in breast cancer cells promoted cell proliferation, colony formation ability, tumor stemness, and tamoxifen resistance. In addition, ZNF528-AS1 overexpression enhanced the migration and invasion abilities of breast cancer cells. ZNF528-AS1 knockdown reduced cell proliferation, tamoxifen resistance, tumor stemness, migration, and invasion. ZNF528-AS1 overexpression might be involved in the regulation of transforming growth factor(TGF-β)signaling pathway. ZNF528-AS1 activated both canonical and non-canonical TGF-β signaling pathways. TGF-β pathway inhibitor SB431542 reversed the proliferation, migration and invasion of breast cancer cells enhanced by ZNF528-AS1 overexpression. Conclusion ZNF528-AS1 can promote tamoxifen resistance and progression of breast cancer, and its mechanism might be related to the activation of TGF-β pathway.

Key words: Breast cancer, ZNF528-AS1, Tamoxifen resistance, Tumor stemness, Progression

中图分类号: 

  • R737.9
[1] Harrod A, Lai CF, Goldsbrough I, et al. Genome engineering for estrogen receptor mutations reveals differential responses to anti-estrogens and new prognostic gene signatures for breast cancer [J]. Oncogene, 2022. doi:0.1038/s41388-022-02483-8.
[2] Battisti NML, Smith IE. Preventing late recurrence in hormone receptor-positive early breast cancer: a review [J]. Eur J Cancer, 2022, 172: 53-64. doi: 10.1016/j.ejca.2022.05.028.
[3] Jeffreys SA, Powter B, Balakrishnar B, et al. Endocrine resistance in breast cancer: the role of estrogen receptor stability [J]. Cells, 2020, 9(9): 2077. doi: 10.3390/cells9092077.
[4] Chen B, Dragomir MP, Yang C, et al. Targeting non-coding RNAs to overcome cancer therapy resistance [J]. Signal Transduct Target Ther, 2022, 7(1): 121. doi: 10.1038/s41392-022-00975-3.
[5] 刘岩, 张曼, 姜朝阳, 等. LncRNA-HOTAIR调控H3K27me3影响巨噬细胞迁移的机制[J]. 山东大学学报(医学版), 2022, 60(6): 1-9. LIU Yan, ZHANG Man, JIANG Chaoyang, et al. Mechanism of LncRNA-HOTAIR regulating macrophage migration through modulating H3K27me3 [J]. Journal of Shandong University(Health Sciences), 2022, 60(6): 1-9.
[6] Lin X, Dinglin X, Cao S, et al. Enhancer-driven lncRNA BDNF-AS induces endocrine resistance and malignant progression of breast cancer through the RNH1/TRIM21/mTOR cascade [J]. Cell Rep, 2020, 31(10): 107753. doi: 10.1016/j.celrep.2020.107753.
[7] Barazetti JF, Jucoski TS, Carvalho TM, et al. From micro to long: non-coding RNAs in tamoxifen resistance of breast cancer cells [J]. Cancers(Basel), 2021, 13(15): 3688. doi: 10.3390/cancers13153688.
[8] Zhang H, Zhang J, Dong L, et al. LncRNA ATXN8OS enhances tamoxifen resistance in breast cancer [J]. Open Med(Wars), 2021, 16(1): 68-80.
[9] Shi Q, Li Y, Li S, et al. LncRNA DILA1 inhibits Cyclin D1 degradation and contributes to tamoxifen resistance in breast cancer [J]. Nat Commun, 2020, 11(1): 5513. doi: 10.1038/s41467-020-19349-w.
[10] Saha T, Lukong KE. Breast cancer stem-like cells in drug resistance: a review of mechanisms and novel therapeutic strategies to overcome drug resistance [J]. Front Oncol, 2022, 12: 856974. doi: 10.3389/fonc.2022.856974.
[11] Mao XD, Wei X, Xu T, et al. Research progress in breast cancer stem cells: characterization and future perspectives [J]. Am J Cancer Res, 2022, 12(7): 3208-3222.
[12] Xu H, Zhang F, Gao X, et al. Fate decisions of breast cancer stem cells in cancer progression [J]. Front Oncol, 2022, 12: 968306. doi: 10.3389/fonc.2022.968306.
[13] Ibragimova M, Tsyganov M, Litviakov N. Tumour stem cells in breast cancer [J]. Int J Mol Sci, 2022, 23(9): 5058. doi: 10.3390/ijms23095058.
[14] Alataki A, Dowsett M. Human epidermal growth factor receptor-2 and endocrine resistance in hormone-dependent breast cancer [J]. Endocr Relat Cancer, 2022, 29(8): R105-R122.
[15] Dong C, Wu J, Chen Y, et al. Activation of PI3K/AKT/mTOR pathway causes drug resistance in breast cancer [J]. Front Pharmacol, 2021, 12: 628690. doi: 10.3389/fphar.2021.628690.
[16] Azuma K, Ikeda K, Suzuki T, et al. TRIM47 activates NF-kappaB signaling via PKC-epsilon/PKD3 stabilization and contributes to endocrine therapy resistance in breast cancer [J]. Proc Natl Acad Sci USA, 2021, 118(35): e2100784118. doi: 10.1073/pnas.2100784118.
[17] Shi X, Yang J, Deng S, et al. TGF-beta signaling in the tumor metabolic microenvironment and targeted therapies [J]. J Hematol Oncol, 2022, 15(1): 135. doi: 10.1186/s13045-022-01349-6.
[18] Takahashi K, Podyma-Inoue KA, Saito M, et al. TGF-beta generates a population of cancer cells residing in G1 phase with high motility and metastatic potential via KRTAP2-3 [J]. Cell Rep, 2022, 40(13): 111411. doi: 10.1016/j.celrep.2022.111411.
[19] Maslankova J, Vecurkovska I, Rabajdova M, et al. Regulation of transforming growth factor-beta signaling as a therapeutic approach to treating colorectal cancer [J]. World J Gastroenterol, 2022, 28(33): 4744-4761.
[20] 刘群, 李丽娜, 刘健, 等. 卵巢癌SOX4介导TGF-β1诱导的上皮间质转化对侵袭转移能力的影响 [J]. 首都医科大学学报, 2022, 43(3): 336-342. LIU Qun, LI Lina, LIU Jian, et al. Effect of SOX4-mediated TGF-β1-induced epithelial-mesenchymal transition on invasion and metastasis in ovarian cancer [J]. Journal of Capital Medical University, 2022, 43(3): 336-342.
[21] Peng P, Zhu H, Liu D, et al. TGFBI secreted by tumor-associated macrophages promotes glioblastoma stem cell-driven tumor growth via integrin alphavbeta5-Src-Stat3 signaling [J]. Theranostics, 2022, 12(9): 4221-4236.
[22] Thambyrajah R, Monteiro R. In the spotlight: the role of TGFbeta signalling in haematopoietic stem and progenitor cell emergence [J]. Biochem Soc Trans, 2022, 50(2): 703-712.
[23] Garcia-Gomez P, Golan I, Dadras MS, et al. NOX4 regulates TGFbeta-induced proliferation and self-renewal in glioblastoma stem cells [J]. Mol Oncol, 2022, 16(9): 1891-1912.
[24] Zhang Z, Xu Y. FZD7 accelerates hepatic metastases in pancreatic cancer by strengthening EMT and stemness associated with TGF-beta/SMAD3 signaling [J]. Mol Med, 2022, 28(1): 82. doi: 10.1186/s10020-022-00509-1.
[25] Wang B, Cao C, Liu X, et al. BRCA1-associated protein inhibits glioma cell proliferation and migration and glioma stem cell self-renewal via the TGF-beta/PI3K/AKT/mTOR signalling pathway [J]. Cell Oncol(Dordr), 2020, 43(2): 223-235.
[26] Rodrigues-Junior DM, Tsirigoti C, Lim SK, et al. Extracellular vesicles and transforming growth factor beta signaling in cancer [J]. Front Cell Dev Biol, 2022, 10: 849938. doi: 10.3389/fcell.2022.849938.
[27] Chan MK, Chung JY, Tang PC, et al. TGF-beta signaling networks in the tumor microenvironment [J]. Cancer Lett, 2022, 550:215925. doi: 10.1016/j.canlet.2022.215925.
[28] Wang Z, Chen J, Wang S, et al. RGS6 suppresses TGF-beta-induced epithelial-mesenchymal transition in non-small cell lung cancers via a novel mechanism dependent on its interaction with SMAD4 [J]. Cell Death Dis, 2022, 13(7): 656. doi: 10.1038/s41419-022-05093-0.
[29] Zou ML, Chen ZH, Teng YY, et al. The Smad dependent TGF-beta and BMP signaling pathway in bone remodeling and therapies [J]. Front Mol Biosci, 2021, 8: 593310. doi: 10.3389/fmolb.2021.593310.
[30] Aykul S, Maust J, Thamilselvan V, et al. Smad2/3 activation regulates Smad1/5/8 signaling via a negative feedback loop to inhibit 3T3-L1 adipogenesis [J]. Int J Mol Sci, 2021, 22(16): 8472. doi: 10.3390/ijms22168472.
[31] Tzavlaki K, Moustakas A. TGF-beta signaling [J]. Biomolecules, 2020, 10(3): 487. doi: 10.3390/biom10030487.
[32] Song X, Wei C, Li X. The signaling pathways associated with breast cancer bone metastasis [J]. Front Oncol, 2022, 12: 855609. doi: 10.3389/fonc.2022.855609.
[33] Chen X, Yan N. Stachydrine inhibits TGF-beta1-induced epithelial-mesenchymal transition in hepatocellular carcinoma cells through the TGF-beta/Smad and PI3K/Akt/mTOR signaling pathways[J]. Anticancer Drugs, 2021, 32(8): 786-792.
[1] 古春青,郭睿思,周勤勤,刘恒辉,巴婉玉,孙士玲,王冰,郑玉玲,吴宿慧. 基于网络药理学和动物实验探讨酸枣仁-远志药对治疗乳腺癌相关性失眠的作用机制[J]. 山东大学学报 (医学版), 2026, 64(1): 99-108.
[2] 刘保国,宋翔,赵晓文,毛亚丽. 血清STAT5B、NKAIN1 mRNA检测在乳腺癌中的应用价值[J]. 山东大学学报 (医学版), 2025, 63(7): 68-74.
[3] 王雪梅,杨豪,宋洋,程世超,张婷婷,王艳春. 抗糖尿病药物与女性恶性肿瘤的因果关联:一项两样本孟德尔随机化分析[J]. 山东大学学报 (医学版), 2025, 63(6): 67-77.
[4] 余之刚,郑超. 乳腺癌多学科诊疗的现状、挑战与创新模式[J]. 山东大学学报 (医学版), 2025, 63(1): 1-9.
[5] 山东省医学会乳腺疾病多学科联合委员会. 乳腺癌多学科协作诊疗山东共识(2024年版)[J]. 山东大学学报 (医学版), 2025, 63(1): 10-16.
[6] 程跃启,王斐,于理想,郑超,余之刚. 曲妥珠单抗致HER2阳性乳腺癌患者心脏毒性的研究进展[J]. 山东大学学报 (医学版), 2025, 63(1): 17-24.
[7] 王敏, 李习平, 檀军, 邱梅, 侯泽宇, 田莹, 罗鸿莹, 范超文, 齐玲, 俞琦, 谢薇. 慢病毒载体介导Gag-Caspase-8诱导三阴性乳腺癌原代细胞凋亡及S期阻滞[J]. 山东大学学报 (医学版), 2025, 63(1): 25-34.
[8] 张洁,张芳芳,王靖楠,李泽宇,宋颖,李娜. circ_0000144在乳腺癌中的表达及其对乳腺癌细胞增殖、迁移和侵袭能力的影响[J]. 山东大学学报 (医学版), 2025, 63(1): 35-42.
[9] 宋雅雯,郭联涛,孔德光,孙圣荣. VTCN1导致HR+乳腺癌预后不良及内分泌治疗耐药[J]. 山东大学学报 (医学版), 2025, 63(1): 43-59.
[10] 刘晶晶,庞婧,赵晓丹,林昕,付敏,陈静静. 基于乳腺X线摄影及DCE-MRI机器学习模型预测乳腺癌新辅助治疗后病理完全缓解:双中心研究[J]. 山东大学学报 (医学版), 2025, 63(1): 60-72.
[11] 孙婧,杨瑞敏,王聪,张月,罗兵. 基于术前超声、炎症指标及超声影像组学联合模型预测乳腺癌腋窝淋巴结转移[J]. 山东大学学报 (医学版), 2025, 63(1): 73-80.
[12] 李永,崔书君,杨飞,张凡,殷晓霞. 基于增强MRI的亚区域影像组学模型可预测乳腺癌患者新辅助化疗后的病理完全反应[J]. 山东大学学报 (医学版), 2025, 63(1): 81-89.
[13] 袁宗怀,潘广晔,迟曰梅,安传国,张永刚. 孟德尔随机化分析低级别浆液性卵巢癌与乳腺癌的因果关系[J]. 山东大学学报 (医学版), 2025, 63(1): 99-107.
[14] 艾宪程,王斐,周文重,于理想,周飞,相玉娟,李亮,余之刚. 山东省38所三甲医院乳腺多学科诊疗现状[J]. 山东大学学报 (医学版), 2025, 63(1): 108-114.
[15] 刘向荣,张新胜,杨荩冉,杨雪艳,刘钊,刘英华. ω-6/ω-3多不饱和脂肪酸比值对不同肿瘤发病风险的系统综述和Meta分析[J]. 山东大学学报 (医学版), 2024, 62(8): 34-48.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
版权所有 © 2018 《山东大学学报(医学版)》编辑部　
地址:山东大学科技期刊社（济南市历城区山大南路27号山东大学中心校区明德楼B座721室） 电话: 0531-88366918 E-mail: xbyxb@sdu.edu.cn
本系统由北京玛格泰克科技发展有限公司设计开发