Zhang Miao, Zhou Xiaolei, Li Zhenzhen, Zhao TianYu, Miao Yu, Liu Sen, Han Qiaoqiao, Wang Libo, Xu Yongdeng, Cui Tao, Wang Ze, Yi Xiulin, Yan Fengying, Wang Xiaoliang
State Key Laboratory of Druggability Evatuation and Systematic Translational Medicine, Tianjin Institute of Pharmaceutical Research, Tianjin, 300301, China.
Saifu Glotope Laboratories Wuxi, Wuxi, 214000, China.
Clin Epigenetics. 2025 Jul 7;17(1):120. doi: 10.1186/s13148-025-01925-w.
To evaluate the efficacy of sorafenib in combination with the DNA methylation inhibitor decitabine (DAC) for the treatment of hepatocellular carcinoma (HCC), and to investigate the mechanism of sorafenib resistance from an epigenetic perspective, aiming to provide new insights and strategies for HCC therapy.
The GEPIA2 database was used to analyze the expression of solute carrier organic anion transporter family member 1B3 (SLCO1B3) in various tumors and adjacent normal tissues. The Kaplan-Meier method was applied to assess the relationship between SLCO1B3 expression and overall survival. The Cancer Genome Atlas (TCGA) Liver Hepatocellular Carcinoma (LIHC) dataset was used to analyze correlations between SLCO1B3 and DNA methyltransferases (DNMTs). Methylation levels of the SLCO1B3 promoter in Hep3B, HepG2, SNU182, and SNU387 cells were determined by bisulfite sequencing PCR. The expression of organic anion transporting polypeptide 1B3 (OATP1B3), encoded by SLCO1B3, was measured by RT-qPCR and Western blot. The effect of sorafenib combined with DAC on Hep3B and HepG2 cells proliferation was dynamically monitored using the Agilent xCELLigence Real-Time Cell Analysis eSight system (RTCA-eSight). The mechanism was further validated in vivo using a Hep3B xenograft model in nude mice. OATP1B3 expression in tumor tissues was examined by immunohistochemistry and Western blot.
HCC patients with high SLCO1B3 expression had significantly better overall survival than those with low expression. SLCO1B3 expression was negatively correlated with DNMTs expression. Compared to other HCC cell lines, Hep3B and HepG2 cells exhibited higher DNA methylation levels and lower OATP1B3 protein expression. DAC treatment upregulated OATP1B3 expression in Hep3B and HepG2 cells. Co-administration of DAC increased sorafenib uptake and enhanced its cytotoxic effect in these cells. In the Hep3B xenograft model, tumor volumes in the combination group were markedly smaller than those in the monotherapy and control groups. OATP1B3 expression was significantly higher in both the combination and DAC-only groups compared to the control and sorafenib-only groups.
DAC promoted OATP1B3 expression by inhibiting SLCO1B3 methylation, thereby enhancing HCC sensitivity to sorafenib. These findings may offer novel therapeutic strategies for the clinical management of HCC.
评估索拉非尼联合DNA甲基化抑制剂地西他滨（DAC）治疗肝细胞癌（HCC）的疗效，并从表观遗传学角度探讨索拉非尼耐药的机制，旨在为HCC治疗提供新的见解和策略。
使用GEPIA2数据库分析溶质载体有机阴离子转运体家族成员1B3（SLCO1B3）在各种肿瘤及相邻正常组织中的表达。采用Kaplan-Meier法评估SLCO1B3表达与总生存期的关系。利用癌症基因组图谱（TCGA）肝细胞癌（LIHC）数据集分析SLCO1B3与DNA甲基转移酶（DNMTs）之间的相关性。通过亚硫酸氢盐测序PCR测定Hep3B、HepG2、SNU182和SNU387细胞中SLCO1B3启动子的甲基化水平。采用RT-qPCR和蛋白质免疫印迹法检测由SLCO1B3编码的有机阴离子转运多肽1B3（OATP1B3）的表达。使用安捷伦xCELLigence实时细胞分析eSight系统（RTCA-eSight）动态监测索拉非尼联合DAC对Hep3B和HepG2细胞增殖的影响。使用Hep3B裸鼠异种移植模型在体内进一步验证其机制。通过免疫组织化学和蛋白质免疫印迹法检测肿瘤组织中OATP1B3的表达。
SLCO1B3高表达的HCC患者总生存期明显优于低表达患者。SLCO1B3表达与DNMTs表达呈负相关。与其他HCC细胞系相比，Hep3B和HepG2细胞表现出更高的DNA甲基化水平和更低的OATP1B3蛋白表达。DAC处理上调了Hep3B和HepG2细胞中OATP1B3的表达。联合使用DAC增加了索拉非尼的摄取并增强了其在这些细胞中的细胞毒性作用。在Hep3B异种移植模型中，联合治疗组的肿瘤体积明显小于单药治疗组和对照组。与对照组和仅使用索拉非尼组相比，联合治疗组和仅使用DAC组中OATP1B3的表达均显著更高。
DAC通过抑制SLCO1B3甲基化促进OATP1B3表达，从而增强HCC对索拉非尼的敏感性。这些发现可能为HCC的临床管理提供新的治疗策略。
