TMEM120B与MYH9结合,通过β1-整合素/FAK-TAZ-mTOR信号轴增强乳腺癌细胞的干性并加速化疗耐受性

IF 6.1 1区 医学 Q1 ONCOLOGY Breast Cancer Research Pub Date : 2024-03-19 DOI:10.1186/s13058-024-01802-z
Ran Hu, Yu Cao, Yuanyuan Wang, Tingting Zhao, Kaibo Yang, Mingwei Fan, Mengyao Guan, Yuekang Hou, Jiao Ying, Xiaowen Ma, Ning Deng, Xun Sun, Yong Zhang, Xiupeng Zhang
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引用次数: 0

摘要

乳腺癌干细胞(CSC)扩增会导致肿瘤进展和化疗耐药性;然而,对CSC多能性的调控仍有待探索。跨膜蛋白120B(TMEM120B)是一种新发现的在人体组织中表达的蛋白,尤其是在恶性组织中;然而,它在CSC扩增中的作用尚未被研究。本研究旨在确定TMEM120B在具有PDZ结合基调的转录辅激活因子(TAZ)介导的CSC扩增和化疗耐药中的作用。研究人员通过生物信息学分析和免疫组化检测,研究了TMEM120B在肺癌、乳腺癌、胃癌、结肠癌和卵巢癌中的表达模式。此外,还对临床病理因素和总生存率进行了评估。接着,研究人员进行了菌落形成试验、MTT试验、EdU试验、transwell试验、伤口愈合试验、流式细胞分析、球体形成试验、Western印迹分析、小鼠异种移植模型分析、RNA测序试验、免疫荧光试验和反转录聚合酶链反应,以研究TMEM120B相互作用对增殖、侵袭、干性、化疗敏感性和整合素/FAK/TAZ/mTOR激活的影响。此外,还进行了液相色谱-串联质谱分析、GST牵引试验和免疫沉淀试验,以评估TMEM120B、肌球蛋白重链9(MYH9)和CUL9之间的相互作用。TMEM120B在肺癌、乳腺癌、胃癌、结肠癌和卵巢癌中表达升高。TMEM120B的表达与TNM分期晚期、淋巴结转移和预后不良呈正相关。TMEM120B的过表达通过激活TAZ-mTOR信号传导促进了乳腺癌细胞的增殖、侵袭和干性。TMEM120B直接与MYH9的线圈盘旋结构域结合,从而加速了焦点粘附(FA)的组装,并促进了TAZ的转位。此外,TMEM120B还以泛素依赖的方式阻止MYH9被CUL9降解,从而稳定了MYH9。过表达 TMEM120B 会增强对多西他赛和多柔比星的耐药性。相反,TMEM120B-∆CCD的过表达会延迟FA的形成,抑制TAZ-mTOR信号转导,减轻化疗耐药性。治疗效果不佳的乳腺癌患者(Miller/Payne 分级 1-2)的 TMEM120B 表达高于治疗效果较好(Miller/Payne 分级 3-5)的患者。我们的研究发现,TMEM120B 与 MYH9 结合并阻止其降解,从而稳定了 MYH9。这种相互作用激活了β1-整合素/FAK-TAZ-mTOR信号轴,维持了干性并加速了化疗耐药性。
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TMEM120B strengthens breast cancer cell stemness and accelerates chemotherapy resistance via β1-integrin/FAK-TAZ-mTOR signaling axis by binding to MYH9
Breast cancer stem cell (CSC) expansion results in tumor progression and chemoresistance; however, the modulation of CSC pluripotency remains unexplored. Transmembrane protein 120B (TMEM120B) is a newly discovered protein expressed in human tissues, especially in malignant tissues; however, its role in CSC expansion has not been studied. This study aimed to determine the role of TMEM120B in transcriptional coactivator with PDZ-binding motif (TAZ)-mediated CSC expansion and chemotherapy resistance. Both bioinformatics analysis and immunohistochemistry assays were performed to examine expression patterns of TMEM120B in lung, breast, gastric, colon, and ovarian cancers. Clinicopathological factors and overall survival were also evaluated. Next, colony formation assay, MTT assay, EdU assay, transwell assay, wound healing assay, flow cytometric analysis, sphere formation assay, western blotting analysis, mouse xenograft model analysis, RNA-sequencing assay, immunofluorescence assay, and reverse transcriptase-polymerase chain reaction were performed to investigate the effect of TMEM120B interaction on proliferation, invasion, stemness, chemotherapy sensitivity, and integrin/FAK/TAZ/mTOR activation. Further, liquid chromatography–tandem mass spectrometry analysis, GST pull-down assay, and immunoprecipitation assays were performed to evaluate the interactions between TMEM120B, myosin heavy chain 9 (MYH9), and CUL9. TMEM120B expression was elevated in lung, breast, gastric, colon, and ovarian cancers. TMEM120B expression positively correlated with advanced TNM stage, lymph node metastasis, and poor prognosis. Overexpression of TMEM120B promoted breast cancer cell proliferation, invasion, and stemness by activating TAZ-mTOR signaling. TMEM120B directly bound to the coil-coil domain of MYH9, which accelerated the assembly of focal adhesions (FAs) and facilitated the translocation of TAZ. Furthermore, TMEM120B stabilized MYH9 by preventing its degradation by CUL9 in a ubiquitin-dependent manner. Overexpression of TMEM120B enhanced resistance to docetaxel and doxorubicin. Conversely, overexpression of TMEM120B-∆CCD delayed the formation of FAs, suppressed TAZ-mTOR signaling, and abrogated chemotherapy resistance. TMEM120B expression was elevated in breast cancer patients with poor treatment outcomes (Miller/Payne grades 1–2) than in those with better outcomes (Miller/Payne grades 3–5). Our study reveals that TMEM120B bound to and stabilized MYH9 by preventing its degradation. This interaction activated the β1-integrin/FAK-TAZ-mTOR signaling axis, maintaining stemness and accelerating chemotherapy resistance.
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来源期刊
Breast Cancer Research
Breast Cancer Research 医学-肿瘤学
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期刊介绍: Breast Cancer Research is an international, peer-reviewed online journal, publishing original research, reviews, editorials and reports. Open access research articles of exceptional interest are published in all areas of biology and medicine relevant to breast cancer, including normal mammary gland biology, with special emphasis on the genetic, biochemical, and cellular basis of breast cancer. In addition to basic research, the journal publishes preclinical, translational and clinical studies with a biological basis, including Phase I and Phase II trials.
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