PDF(Pubmed)
Abstract：
Cholangiocarcinoma (CCA) is an aggressive cancer originating from bile duct epithelium, particularly prevalent in Asian countries with liver fluke infections. Current chemotherapy for CCA often fails due to drug resistance, necessitating novel anticancer agents. This study investigates the potential of 5\'-deoxy-5\'-methylthioadenosine (MTA), a naturally occurring nucleoside, against CCA. While MTA has shown promise against various cancers, its effects on CCA remain unexplored. We evaluated MTA\'s anticancer activity in CCA cell lines and drug-resistant sub-lines, assessing cell viability, migration, invasion, and apoptosis. The potential anticancer mechanisms of MTA were explored through proteomic analysis using LC-MS/MS and bioinformatic analysis. The results show a dose-dependent reduction in CCA cell viability, with enhanced effects on cancer cells compared to normal cells. Moreover, MTA inhibits growth, induces apoptosis, and suppresses cell migration and invasion. Additionally, MTA enhanced the anticancer effects of gemcitabine on drug-resistant CCA cells. Proteomics revealed the down-regulation of multiple proteins by MTA, affecting various molecular functions, biological processes, and cellular components. Network analysis highlighted MTA\'s role in inhibiting proteins related to mitochondrial function and energy derivation, crucial for cell growth and survival. Additionally, MTA suppressed proteins involved in cell morphology and cytoskeleton organization, important for cancer cell motility and metastasis. Six candidate genes, including ZNF860, KLC1, GRAMD1C, MAMSTR, TANC1, and TTC13, were selected from the top 10 most down-regulated proteins identified in the proteomics results and were subsequently verified through RT-qPCR. Further, KLC1 protein suppression by MTA treatment was confirmed through Western blotting. Additionally, based on TCGA data, KLC1 mRNA was found to be upregulated in the tissue of CCA patients compared to that of normal adjacent tissues. In summary, MTA shows promising anticancer potential against CCA by inhibiting growth, inducing apoptosis, and suppressing migration and invasion, while enhancing gemcitabine\'s effects. Proteomic analysis elucidates possible molecular mechanisms underlying MTA\'s anticancer activity, laying the groundwork for future research and development of MTA as a treatment for advanced CCA.
摘要：
胆管癌（CCA）是起源于胆管上皮的侵袭性癌症，在亚洲国家尤其普遍的肝吸虫感染。目前用于CCA的化疗通常由于耐药性而失败，需要新的抗癌剂。本研究调查了5'-脱氧-5'-甲硫腺苷（MTA）的潜力，一种天然存在的核苷，反对CCA。虽然MTA已经显示出对抗各种癌症的希望，其对CCA的影响仍未被探索。我们评估了MTA在CCA细胞系和耐药亚系中的抗癌活性，评估细胞活力，迁移,入侵,和凋亡。通过使用LC-MS/MS的蛋白质组学分析和生物信息学分析，探索了MTA的潜在抗癌机制。结果显示CCA细胞活力的剂量依赖性降低，与正常细胞相比，对癌细胞的作用增强。此外，MTA抑制生长,诱导细胞凋亡，并抑制细胞迁移和侵袭。此外,MTA增强吉西他滨对耐药CCA细胞的抗癌作用。蛋白质组学揭示了MTA对多种蛋白质的下调，影响各种分子功能，生物过程，和细胞成分。网络分析强调MTA在抑制与线粒体功能和能量衍生相关的蛋白质中的作用，对细胞生长和存活至关重要。此外,MTA抑制参与细胞形态和细胞骨架组织的蛋白质，重要的癌细胞运动和转移。六个候选基因,包括ZNF860，KLC1，GRAMD1C，妈妈,TANC1和TTC13选自蛋白质组学结果中鉴定的前10种最下调的蛋白质,随后通过RT-qPCR验证。Further,通过蛋白质印迹证实MTA处理对KLC1蛋白的抑制。此外,基于TCGA数据，与正常邻近组织相比，CCA患者的组织中KLC1mRNA被上调。总之,MTA通过抑制生长显示出对CCA的有希望的抗癌潜力，诱导细胞凋亡，抑制迁移和入侵，同时增强吉西他滨的效果。蛋白质组学分析阐明了MTA抗癌活性潜在的分子机制，为未来MTA作为先进CCA治疗方法的研究和发展奠定基础。
