School of Pharmacy, Guilin Medical University, Guilin, PR China.
School of Basic Medical Sciences, Youjiang Medical University for Nationalities, Baise, PR China.
Pharm Biol. 2022 Dec;60(1):1566-1577. doi: 10.1080/13880209.2022.2106251.
Urolithin A (UroA) can inhibit the growth of many human cancer cells, but it has not be reported if UroA inhibits nasopharyngeal carcinoma (NPC) cells.
To explore the inhibitory effect of UroA on NPC and potential mechanism .
RNA-sequencing-based mechanistic prediction was conducted by comparing KEGG enrichment of 40 μM UroA-treated for 24 h with untreated CNE2 cells. The untreated cells were selected as control. After NPC cells were treated with 20-60 μM UroA, proliferation, migration and invasion of were measured by colony formation, wound healing and transwell experiments. Apoptosis, mitochondrial membrane potential (MMP), reactive oxygen species (ROS) were measured by flow cytometry, Hoechst 33342, Rhodamine 123, JC-1 staining and ROS assay methods, respectively. Gene and protein expression were measured by RT-qPCR and Western blotting assay.
RNA-sequencing and KEGG enrichment revealed UroA mainly altered the ECM receptor interaction pathway. UroA inhibited cells proliferation, epithelial-mesenchymal-transition pathway, migration and invasion with IC values of 34.72 μM and 44.91 μM, induced apoptosis, MMP depolarization and increase ROS content at a concentration of 40 μM. UroA up-regulated E-cadherin, Bax/Bcl-2, c-caspase-3 and PARP proteins, while inhibiting COL4A1, MMP2, MMP9, N-cadherin, Vimentin and Snail proteins at 20-60 μM. Moreover, co-treatment of UroA (40 μM) and NAC (5 mM) could reverse the effect of UroA on apoptosis-related proteins.
RNA-sequencing technology based on bioinformatic analyses may be applicable for studiying the mechanism of drugs for tumour treatment.
尿石素 A（UroA）可以抑制许多人类癌细胞的生长，但尚未有报道表明 UroA 是否可以抑制鼻咽癌（NPC）细胞。
探索 UroA 对 NPC 的抑制作用及其潜在机制。
通过比较 40μM UroA 处理 24 小时与未处理的 CNE2 细胞的 KEGG 富集，进行基于 RNA 测序的机制预测。未处理的细胞被选为对照。用 20-60μM UroA 处理 NPC 细胞后，通过集落形成、划痕愈合和 Transwell 实验测量细胞增殖、迁移和侵袭。通过流式细胞术、Hoechst 33342、罗丹明 123、JC-1 染色和 ROS 测定方法分别测量细胞凋亡、线粒体膜电位（MMP）和活性氧（ROS）。通过 RT-qPCR 和 Western blot 测定基因和蛋白表达。
RNA 测序和 KEGG 富集表明 UroA 主要改变了 ECM 受体相互作用途径。UroA 在 34.72μM 和 44.91μM 时抑制细胞增殖、上皮-间充质转化途径、迁移和侵袭，并在 40μM 时诱导细胞凋亡、MMP 去极化和增加 ROS 含量。UroA 在 20-60μM 时上调 E-cadherin、Bax/Bcl-2、c-caspase-3 和 PARP 蛋白，同时抑制 COL4A1、MMP2、MMP9、N-cadherin、Vimentin 和 Snail 蛋白。此外，UroA（40μM）和 NAC（5mM）联合处理可逆转 UroA 对凋亡相关蛋白的作用。
基于生物信息学分析的 RNA 测序技术可能适用于研究肿瘤治疗药物的作用机制。
