Department of Urology, Deyang People's Hospital, Deyang, 618000, China.
Department of Urology, School of Medicine, Sichuan Provincial People's Hospital, Chengdu, 610072, China.
Amino Acids. 2024 Sep 30;56(1):57. doi: 10.1007/s00726-024-03419-0.
Function of HECTD2 in renal cell carcinoma malignant progression is undefined. Molecular mechanism behind anti-cancer effects of veratric acid (VA) from traditional Chinese medicine (TCM) is underexplored. The Cancer Genome Atlas was leveraged to study HECTD2 expression in renal cell carcinoma and its relationship with histological grading. Kaplan-Meier survival analysis was performed. HECTD2 expression was detected in cancer cells and tissues via qRT-PCR and immunohistochemistry. GPX4 and SLC7A11 expression in tumor samples with high or low HECTD2 expression was examined by immunohistochemistry, cell viability by CCK8, cell proliferation by colony formation assay, lipid ROS and mitochondrial superoxide levels by flow cytometry, Fe and MDA content by assay kits, and GPX4 and SLC7A11 proteins by western blot. SeeSAR software screened TCM small molecule compounds with highest affinity to HECTD2, confirmed with cellular thermal shift assay. VA IC was measured by CCK8. Xenograft model was developed and treated with VA. Tumor size and weight were monitored, with immunohistochemistry to detect HECTD2 expression in tumors and assess ferroptosis-related markers. HECTD2 was overexpressed in tumor tissues and cells, which positively correlated with histological grading. HECTD2 depletion inhibited cell vitality and proliferation, raised intracellular lipid ROS, mitochondrial superoxide, Fe, and MDA. HECTD2 was a target with highest VA affinity. In vitro and vivo experiments concurred that VA treatment hindered malignancy of renal cell carcinoma and enhanced its susceptibility to ferroptosis. HECTD2 supports ferroptosis resistance in renal cell carcinoma, but VA, through its targeting of HECTD2, initiates ferroptosis, showcasing its anti-cancer efficacy.
HECTD2 在肾细胞癌恶性进展中的功能尚未确定。来自中药的原儿茶酸 (VA) 的抗癌作用的分子机制尚未得到充分探索。利用癌症基因组图谱研究肾细胞癌中 HECTD2 的表达及其与组织学分级的关系。进行 Kaplan-Meier 生存分析。通过 qRT-PCR 和免疫组织化学检测癌细胞和组织中的 HECTD2 表达。通过免疫组织化学检测高或低 HECTD2 表达肿瘤样本中的 GPX4 和 SLC7A11 表达，通过 CCK8 检测细胞活力，通过集落形成试验检测细胞增殖，通过流式细胞术检测脂质 ROS 和线粒体超氧化物水平，通过试剂盒检测 Fe 和 MDA 含量，通过 Western blot 检测 GPX4 和 SLC7A11 蛋白。SeeSAR 软件筛选与 HECTD2 亲和力最高的中药小分子化合物，并用细胞热转移试验确认。通过 CCK8 测量 VA 的 IC。建立异种移植模型并用 VA 治疗。监测肿瘤大小和重量，通过免疫组织化学检测肿瘤中 HECTD2 的表达并评估铁死亡相关标志物。HECTD2 在肿瘤组织和细胞中过度表达，与组织学分级呈正相关。HECTD2 耗竭抑制细胞活力和增殖，增加细胞内脂质 ROS、线粒体超氧化物、Fe 和 MDA。HECTD2 是与 VA 亲和力最高的靶标。体内外实验均表明，VA 治疗抑制肾细胞癌的恶性程度并增强其对铁死亡的敏感性。HECTD2 支持肾细胞癌的铁死亡抵抗，但 VA 通过靶向 HECTD2 引发铁死亡，展示其抗癌功效。
