Khan Hanif, Ni Zhengzhong, Feng Hai, Xing Yaqi, Wu Xuejun, Huang Danmei, Chen Ling, Niu Yongdong, Shi Ganggang
Department of Pharmacology, Shantou University Medical College, No. 22 Xinling Road, Shantou 515041, Guangdong, China.
Shanghai Shuguang Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai, China.
Phytomedicine. 2021 Oct;91:153706. doi: 10.1016/j.phymed.2021.153706. Epub 2021 Aug 14.
Hepatocellular carcinoma (HCC) is one of the most common cause of cancer-related death worldwide. Curcumin (C) has been extensively investigated in different types of malignancies, including hepatocellular carcinoma, but its physicochemical properties have significantly influenced its clinical use. Several approaches are being explored to enhance curcumin's therapeutic response, including its combination with various drugs.
This study aimed to evaluate the anti-tumor effect of curcumin (C) in combination with F2 (N-n-butyl haloperidol iodide) on hepatocellular carcinoma and its potential underlying mechanism in vitro and in vivo.
Cell proliferation was evaluated by CCK-8 and colony formation assays, and apoptosis was measured by flow cytometry. The migratory and invasive abilities of Hep3B and SMMC-7721 cells were measured by wound-healing and matrigel transwell assays. In order to investigate the molecular pathways, various experiments such as western blotting, qPCR, RNA-seq, immunostaining and transfection were performed. To evaluate the anti-HCC effects in vivo, a xenograft tumor model was used.
Our findings showed that the combination of curcumin (C) & F2 (F2C) strongly inhibited malignant proliferation and migration in SMMC-7721 and Hep3B cells. The F2C treatment downregulates enhancer of zeste homolog 2 (EZH2) transcription and protein expression, which is key epigenetic regulator responsible for HCC development. Moreover, the inhibition of EZH2 by F2C led to Wnt/β-catenin signaling inhibition by decreasing tri-methylation of histone H3 at lysine 27 (H3K27me3) and long non-coding RNA H19 expression. The inhibition of F2C was associated with the suppression of tumorigenicity in xenograft HCC models.
These findings suggested that, F2C inhibited HCC formation, migration and its modulatory mechanism seemed to be associated with downregulation of EZH2, silencing Wnt/β-catenin signaling by interacting with H19, suggesting that F2C may be a promising drug in the clinical treatment of HCC.
肝细胞癌（HCC）是全球癌症相关死亡的最常见原因之一。姜黄素（C）已在包括肝细胞癌在内的不同类型恶性肿瘤中得到广泛研究，但其物理化学性质显著影响了其临床应用。目前正在探索多种方法来增强姜黄素的治疗反应，包括将其与各种药物联合使用。
本研究旨在评估姜黄素（C）与F2（N-正丁基碘化卤哌啶）联合应用对肝细胞癌的抗肿瘤作用及其在体外和体内潜在的作用机制。
通过CCK-8和集落形成试验评估细胞增殖，通过流式细胞术检测细胞凋亡。采用伤口愈合试验和基质胶Transwell试验检测Hep3B和SMMC-7721细胞的迁移和侵袭能力。为了研究分子途径，进行了蛋白质免疫印迹、qPCR、RNA测序、免疫染色和转染等各种实验。为了评估体内抗肝癌作用，使用了异种移植肿瘤模型。
我们的研究结果表明，姜黄素（C）与F2（F2C）联合使用可强烈抑制SMMC-7721和Hep3B细胞的恶性增殖和迁移。F2C处理下调了zeste同源物2（EZH2）的转录和蛋白表达，EZH2是负责HCC发生发展的关键表观遗传调节因子。此外，F2C对EZH2的抑制通过降低组蛋白H3赖氨酸27（H3K27me3）的三甲基化和长链非编码RNA H19的表达导致Wnt/β-连环蛋白信号通路受到抑制。F2C的抑制作用与异种移植肝癌模型中肿瘤发生的抑制相关。
这些研究结果表明，F2C抑制HCC的形成、迁移，其调节机制似乎与EZH2的下调有关，通过与H19相互作用使Wnt/β-连环蛋白信号通路沉默，提示F2C可能是HCC临床治疗中有前景的药物。
