Department of Biology, University of Alabama at Birmingham, 902 14th Street South, Birmingham, AL, 35294, USA.
Department of Microbiology, Heersink School of Medicine, University of Alabama at Birmingham, Birmingham, AL, 35205, USA.
Sci Rep. 2024 May 27;14(1):12091. doi: 10.1038/s41598-024-62084-1.
Estrogen receptor-negative [ER(-)] mammary cancer is the most aggressive type of breast cancer (BC) with higher rate of metastasis and recurrence. In recent years, dietary prevention of BC with epigenetically active phytochemicals has received increased attention due to its feasibility, effectiveness, and ease of implementation. In this regard, combinatorial phytochemical intervention enables more efficacious BC inhibition by simultaneously targeting multiple tumorigenic pathways. We, therefore, focused on investigation of the effect of sulforaphane (SFN)-rich broccoli sprouts (BSp) and withaferin A (WA)-rich Ashwagandha (Ash) combination on BC prevention in estrogen receptor-negative [ER(-)] mammary cancer using transgenic mice. Our results indicated that combinatorial BSp + Ash treatment significantly reduced tumor incidence and tumor growth (~ 75%) as well as delayed (~ 21%) tumor latency when compared to the control treatment and combinatorial BSp + Ash treatment was statistically more effective in suppressing BC compared to single BSp or Ash intervention. At the molecular level, the BSp and Ash combination upregulated tumor suppressors (p53, p57) along with apoptosis associated proteins (BAX, PUMA) and BAX:BCL-2 ratio. Furthermore, our result indicated an expressional decline of epigenetic machinery HDAC1 and DNMT3A in mammary tumor tissue because of combinatorial treatment. Interestingly, we have reported multiple synergistic interactions between BSp and Ash that have impacted both tumor phenotype and molecular expression due to combinatorial BSp and Ash treatment. Our RNA-seq analysis results also demonstrated a transcriptome-wide expressional reshuffling of genes associated with multiple cell-signaling pathways, transcription factor activity and epigenetic regulations due to combined BSp and Ash administration. In addition, we discovered an alteration of gut microbial composition change because of combinatorial treatment. Overall, combinatorial BSp and Ash supplementation can prevent ER(-) BC through enhanced tumor suppression, apoptosis induction and transcriptome-wide reshuffling of gene expression possibly influencing multiple cell signaling pathways, epigenetic regulation and reshaping gut microbiota.
雌激素受体阴性（ER(-)）乳腺癌是最具侵袭性的乳腺癌（BC）类型，具有更高的转移和复发率。近年来，由于其可行性、有效性和易于实施，饮食预防 BC 的方法引起了人们的广泛关注，其中包括使用具有表观遗传活性的植物化学物质。在这方面，联合植物化学干预可以通过同时针对多个致癌途径来更有效地抑制 BC。因此，我们专注于研究富含萝卜硫素（SFN）的西兰花芽（BSp）和富含醉茄内酯 A（WA）的 Ashwagandha（Ash）组合对雌激素受体阴性（ER(-)）乳腺癌的预防作用，使用转基因小鼠进行研究。我们的结果表明，与对照组相比，联合 BSp+Ash 治疗显著降低了肿瘤发生率和肿瘤生长（75%），并延迟了（21%）肿瘤潜伏期，并且与单独的 BSp 或 Ash 干预相比，联合 BSp+Ash 治疗在抑制 BC 方面更有效。在分子水平上，BSp 和 Ash 联合使用上调了肿瘤抑制因子（p53、p57）以及与细胞凋亡相关的蛋白（BAX、PUMA）和 BAX:BCL-2 比值。此外，我们的结果表明，由于联合治疗，乳腺癌组织中的表观遗传机制 HDAC1 和 DNMT3A 的表达下降。有趣的是，我们报道了 BSp 和 Ash 之间的多种协同相互作用，由于联合 BSp 和 Ash 治疗，这些协同作用影响了肿瘤表型和分子表达。我们的 RNA-seq 分析结果还表明，由于联合 BSp 和 Ash 给药，与多个细胞信号通路、转录因子活性和表观遗传调控相关的基因在全转录组水平上的表达发生了重新排列。此外，我们发现由于联合治疗，肠道微生物组成发生了变化。总的来说，联合 BSp 和 Ash 补充可以通过增强肿瘤抑制、诱导细胞凋亡和全转录组基因表达的重新排列来预防 ER(-)BC，可能影响多个细胞信号通路、表观遗传调控和重塑肠道微生物群。
