Gonzalez Suarez Narjara, Fernandez-Marrero Yuniel, Hébert Mathieu P A, Roy Marie-Eve, Boudreau Luc H, Annabi Borhane
Laboratoire d'Oncologie Moléculaire, Département de Chimie, Université du Québec À Montréal and CERMO-FC, C.P. 8888, Succ. Centre-Ville, Montreal, QC, H3C 3P8, Canada.
Cell Biology Department, NuChem Sciences, Montreal, QC, H4R 2N6, Canada.
Cancer Cell Int. 2023 Oct 13;23(1):240. doi: 10.1186/s12935-023-03087-2.
Triple-negative breast cancer (TNBC) cells' secretome can induce a pro-inflammatory phenotype in human adipose-derived mesenchymal stem cells (hADMSC). This can be prevented by the green tea polyphenol epigallocatechin-3-gallate (EGCG). The impact of EGCG on the paracrine regulation that the extracellular vesicles (EVs) specifically exert within the TNBC secretome remains unknown.
EVs were obtained from a TNBC-derived serum-starved MDA-MB-231 cell model treated or not with EGCG under normoxic or hypoxic (< 1% O) culture conditions. RNA-Seq analysis was used to assess the EVs' genetic content. The modulation of inflammatory and senescence markers in hADMSC was evaluated by RT-qPCR using cDNA arrays and validated by immunoblotting. A protein profiler phospho-kinase array was used to explore signaling pathways.
While hypoxic culture conditions did not significantly alter the genetic content of MDA-MB-231-secreted EVs, the addition of EGCG significantly modified EVs genetic material at low oxygen tension. Gene expression of cancer-associated adipocyte pro-inflammatory markers CXCL8, CCL2 and IL-1β was increased in hADMSC treated with EVs. Concomitantly, EVs isolated from MDA-MB-231 treated with EGCG (EGCG-EVs) downregulated CCL2 and IL-1β, while inducing higher expression of CXCL8 and IL-6 levels. EVs activated CHK-2, c-Jun, AKT and GSK-3β signaling pathways in hADMSC, whereas EGCG-EVs specifically reduced the latter two as well as the serum starvation-induced senescence markers p21 and β-galactosidase. Finally, the mitochondrial content within the TNBC cells-derived EVs was found reduced upon EGCG treatment.
This proof of concept study demonstrates that the chemopreventive properties of diet-derived polyphenols may efficiently target the paracrine regulation that TNBC cells could exert upon their surrounding adipose tissue microenvironment.
三阴性乳腺癌（TNBC）细胞的分泌组可诱导人脂肪来源间充质干细胞（hADMSC）产生促炎表型。绿茶多酚表没食子儿没食子酸酯（EGCG）可预防这种情况。EGCG对细胞外囊泡（EVs）在TNBC分泌组中特异性发挥的旁分泌调节的影响尚不清楚。
在常氧或低氧（<1%O）培养条件下，从经或未经EGCG处理的TNBC来源的血清饥饿MDA-MB-231细胞模型中获取EVs。RNA测序分析用于评估EVs的遗传内容。使用cDNA阵列通过RT-qPCR评估hADMSC中炎症和衰老标志物的调节，并通过免疫印迹进行验证。使用蛋白质磷酸激酶阵列探索信号通路。
虽然低氧培养条件未显著改变MDA-MB-231分泌的EVs的遗传内容，但在低氧张力下添加EGCG可显著改变EVs的遗传物质。用EVs处理的hADMSC中，癌症相关脂肪细胞促炎标志物CXCL8、CCL2和IL-1β的基因表达增加。同时，从用EGCG处理的MDA-MB-231中分离的EVs（EGCG-EVs）下调CCL2和IL-1β，同时诱导CXCL8和IL-6水平的更高表达。EVs激活hADMSC中的CHK-2、c-Jun、AKT和GSK-3β信号通路，而EGCG-EVs特异性降低后两者以及血清饥饿诱导的衰老标志物p21和β-半乳糖苷酶。最后，发现EGCG处理后TNBC细胞来源的EVs中的线粒体含量减少。
这项概念验证研究表明，饮食来源多酚的化学预防特性可能有效地靶向TNBC细胞对其周围脂肪组织微环境施加的旁分泌调节。
