Yang Wei-Chung Vivian, Lin Yen-Yi, Ding Jeak Ling, Hung Chin-Sheng, Nguyen Phung-Anh, Zhang Bo-Xiang, Hsieh Tsung-Han, Chang Shu-Chun
The Ph.D. Program for Translational Medicine, College for Medical Science and Technology, Taipei Medical University, Taipei, 110, Taiwan.
International Ph.D. Program for Translational Science, College of Medical Science and Technology, Taipei Medical University, Taipei, 110, Taiwan.
Cell Mol Biol Lett. 2025 Jun 29;30(1):75. doi: 10.1186/s11658-025-00752-4.
Benzodiazepines (Diazepam) and related Z-drugs (Zolpidem), henceforth referred to as BZDRs, are widely used for clinical treatment of insomnia and anxiety disorders. BZDRs act on GABA type A receptors to inhibit neurotransmitters. We previously demonstrated that prolonged clinical use of BZDRs exacerbates the risk of breast cancer (BRCA).
By biomedical, health informatics platform analyses and in vivo studies, we explored clinical association between BZDR usage and BRCA development and advancement. Furthermore, by retrospective studies on patient clinical data and in vitro empirical analyses of the impact of BZDR on BRCA cells, and together with ingenuity pathway analysis (IPA) analyses, we validated the signaling pathways and identified potential intermolecular crosstalk involved.
Clinical data showed that BRCA patients on long term treatment with BZDRs suffered increased mortality rate (p = 0.034). Studies on patient samples indicated that among 16 GABA receptors examined, GABRA3 (a pro-tumorigenic player) was significantly upregulated by BZDRs, which advanced BRCA disease. To support our clinical findings, we examined in vivo, the impact of BZDRs on BRCA advancement using MDA-MB231 cells to mediate metastasis in mice model. Our results show that BZDRs indeed promoted cancer advancement to the lungs and localized in the tibia. Using BRCA cell lines, we revealed the molecular-cellular effects of prolonged treatment with BZDRs in vitro. We showed significant metastasis indicated by increased cancer cell migration and invasion, which correlated well with our clinical observations. We discovered that BZDR-mediated GABRA3 stimulation was associated with downregulation of anti-tumorigenic extracellular matrix (ECM) molecules (S100B, COL6A6 and VIT) and upregulation of pro-tumorigenic FBN3 in BRCA cells. Notably, GABRA3-shRNA and GABRA3-CRISPR/Cas9 disrupted the abovementioned dynamics dramatically and suppressed BRCA cell invasion induced by BZDRs. Bioinformatics analyses highlighted molecular pathways showing interplay between GABRA3 and ECMs, which presumably exacerbated BZDR-induced BRCA progression via immune modulators.
Long-term clinical use of BZDRs significantly increased the mortality rate of BRCA patients. We provide in vivo and in vitro evidence confirming that BZDRs promote BRCA advancement. We revealed that BZDR-mediated BRCA signaling pathways through GABRA3-ECMs, which promotes metastasis, probably through immune modulation and changes in the tumor microenvironment.
苯二氮䓬类药物（地西泮）及相关的Z类药物（唑吡坦），以下简称BZDRs，被广泛用于失眠和焦虑症的临床治疗。BZDRs作用于A型γ-氨基丁酸（GABA）受体以抑制神经递质。我们之前证明，长期临床使用BZDRs会增加患乳腺癌（BRCA）的风险。
通过生物医学、健康信息学平台分析及体内研究，我们探索了BZDRs使用与BRCA发生和进展之间的临床关联。此外，通过对患者临床数据的回顾性研究以及BZDRs对BRCA细胞影响的体外实证分析，并结合 Ingenuity 通路分析（IPA），我们验证了信号通路并确定了潜在的分子间串扰。
临床数据显示，长期接受BZDRs治疗的BRCA患者死亡率增加（p = 0.034）。对患者样本的研究表明，在检测的16种GABA受体中，GABRA3（一种促肿瘤发生因子）被BZDRs显著上调，这促进了BRCA疾病的发展。为支持我们的临床发现，我们在体内使用MDA-MB231细胞介导小鼠模型中的转移，研究了BZDRs对BRCA进展的影响。我们的结果表明，BZDRs确实促进了癌症向肺部转移并在胫骨中定位。使用BRCA细胞系，我们在体外揭示了长期用BZDRs治疗的分子细胞效应。我们显示癌细胞迁移和侵袭增加表明有显著的转移，这与我们的临床观察结果高度相关。我们发现BZDR介导的GABRA3刺激与BRCA细胞中抗肿瘤细胞外基质（ECM）分子（S100B、COL6A6和VIT）的下调以及促肿瘤发生的FBN3的上调有关。值得注意的是，GABRA3短发夹RNA（shRNA）和GABRA3-CRISPR/Cas9显著破坏了上述动态变化并抑制了BZDRs诱导的BRCA细胞侵袭。生物信息学分析突出了显示GABRA3与ECM之间相互作用的分子通路，这可能通过免疫调节剂加剧了BZDR诱导的BRCA进展。
长期临床使用BZDRs显著增加了BRCA患者的死亡率。我们提供了体内和体外证据，证实BZDRs促进BRCA进展。我们揭示了BZDR通过GABRA3-ECM介导的BRCA信号通路，这可能通过免疫调节和肿瘤微环境的变化促进转移。
