Key Laboratory of Integrated Oncology and Intelligent Medicine of Zhejiang Province, Department of Hepatobiliary and Pancreatic Surgery, Affiliated Hangzhou First People's Hospital, Zhejiang University School of Medicine, Hangzhou, 310006, People's Republic of China.
Westlake Laboratory of Life Sciences and Biomedicine, Hangzhou, 310024, People's Republic of China.
Cell Commun Signal. 2023 Aug 3;21(1):193. doi: 10.1186/s12964-023-01194-1.
Sorafenib resistance greatly reduces the efficacy of treatments in advanced hepatocellular carcinoma (HCC) patients, but the underlying mechanisms are not thoroughly understood. All-trans retinoic acid (ATRA), an anti-leukaemia agent, has attracted considerable attention due to its role in sensitizing cells to other anticancer treatments. We aimed to investigate the combined effect of ATRA and Sorafenib on HCC and the underlying mechanisms.
CCK-8, cell sphere formation, trans-well migration, and wound-healing assays were used to analyse the biological behaviours of HCC cells in vitro. Western blotting and qRT-PCR analysis were conducted to measure the expression of p21 activated kinase 1 (PAK1) and phospho-p21 activated kinase 1 (pPAK1). Xenograft models were established to confirm the synergistic effects of ATRA and Sorafenib in vivo. TUNEL assays and immunohistochemistry were utilized to determine apoptosis, proliferation, PAK1 and pPAK1 levels in tumour tissues.
We observed that PAK1 was overexpressed in HCC, and its expression was negatively correlated with the survival of patients. PAK1 promoted the proliferation, self-renewal and epithelial-mesenchymal transition of HCC cells. Correlation analysis indicated that the IC of Sorafenib was positively correlated with the level of pPAK1 in HCC cell lines. ATRA inhibited the progression of HCC and sensitized HCC response to Sorafenib by downregulation of PAK1, as shown by the calculated coefficient of drug interaction and the data obtained from xenograft models.
Our findings indicated that instead of treatment with Sorafenib alone, the combination of ATRA and Sorafenib provides a more effective treatment for HCC patients. Video Abstract.
索拉非尼耐药极大地降低了晚期肝细胞癌（HCC）患者的治疗效果，但其中的机制尚不完全清楚。全反式维甲酸（ATRA）作为一种抗白血病药物，因其能够增强细胞对其他抗癌治疗的敏感性而受到广泛关注。我们旨在研究 ATRA 和索拉非尼联合应用于 HCC 的效果及其潜在机制。
使用 CCK-8、细胞球形成、Transwell 迁移和划痕愈合实验分析 HCC 细胞的体外生物学行为。Western blot 和 qRT-PCR 分析用于测量 p21 激活激酶 1（PAK1）和磷酸化 p21 激活激酶 1（pPAK1）的表达。建立异种移植模型以在体内证实 ATRA 和索拉非尼的协同作用。TUNEL 检测和免疫组化用于确定肿瘤组织中的细胞凋亡、增殖、PAK1 和 pPAK1 水平。
我们观察到 PAK1 在 HCC 中过表达，其表达与患者的生存呈负相关。PAK1 促进了 HCC 细胞的增殖、自我更新和上皮-间充质转化。相关性分析表明，索拉非尼的 IC 与 HCC 细胞系中 pPAK1 的水平呈正相关。ATRA 通过下调 PAK1 抑制 HCC 的进展并增强 HCC 对索拉非尼的敏感性，这一点可以从计算的药物相互作用系数和异种移植模型的数据中得到证实。
我们的研究结果表明，与单独使用索拉非尼相比，ATRA 和索拉非尼的联合应用为 HCC 患者提供了更有效的治疗方法。
