Wu Si-Han, Xiao Meng-Chao, Liu Fang, Hong Huan-Yu, Ding Chen-Hong, Zhang Xin, Xie Wei-Fen
Department of Gastroenterology, Changzheng Hospital, Naval Medical University, Shanghai, China.
Department of Gastroenterology, Shanghai East Hospital, School of Medicine, Tongji University, Shanghai, 200092, China.
Discov Oncol. 2024 Dec 5;15(1):752. doi: 10.1007/s12672-024-01661-2.
Hepatocyte nuclear factor 4α (HNF4α) is a key regulator of hepatocyte function and has a strong therapeutic effect on hepatocellular carcinoma (HCC) by inducing the differentiation of hepatoma cell into hepatocytes. Our previous study showed that Tribbles homolog 3 (TRIB3) directly interacts with and promotes the degradation of HNF4α in non-alcoholic fatty liver disease (NAFLD). Disrupting the TRIB3-HNF4α interaction by a cell-permeating peptide, called P-T3H2, stabilized HNF4α protein. This study aimed to assess the anti-tumor impact of P-T3H2 in HCC.
The expression of TRIB3 and HNF4α was evaluated using western blot and immunohistochemistry (IHC). Hepatic functions and cellular senescence of HCC cells were evaluated through periodic acid-Schiff (PAS) staining, acetylated low-density lipoprotein (ac-LDL) uptake and senescence-associated β-galactosidase (SA-β-gal) activity staining, respectively. RNA-Seq analysis was performed to identify differentially expressed genes in Huh7 cells treated with P-T3H2. The impact of P-T3H2 on HCC malignancy was assessed in vitro and in vivo.
TRIB3 exhibited a negative correlation with HNF4α in both human and mouse HCC tissues. The administration of P-T3H2 significantly inhibited the malignancy of HCC cells. Additionally, P-T3H2 stabilized HNF4α protein and facilitated the restoration of hepatic functions and the cellular senescence in HCC cells. RNA-Seq analysis demonstrated that P-T3H2 enhanced the transcriptional activity of HNF4α in HCC. Furthermore, P-T3H2 effectively suppressed the carcinogenesis and progression of HCC in mice.
P-T3H2 suppressed HCC progression through the stabilization of HNF4α protein and may be a promising therapeutic candidate for clinical application in the treatment of HCC.
肝细胞核因子4α（HNF4α）是肝细胞功能的关键调节因子，通过诱导肝癌细胞分化为肝细胞，对肝细胞癌（HCC）具有强大的治疗作用。我们之前的研究表明，在非酒精性脂肪性肝病（NAFLD）中，Tribbles同源物3（TRIB3）直接与HNF4α相互作用并促进其降解。一种名为P-T3H2的细胞穿透肽破坏TRIB3-HNF4α相互作用，可使HNF4α蛋白稳定。本研究旨在评估P-T3H2对HCC的抗肿瘤作用。
采用蛋白质免疫印迹法和免疫组织化学（IHC）法评估TRIB3和HNF4α的表达。分别通过过碘酸希夫（PAS）染色、乙酰化低密度脂蛋白（ac-LDL）摄取和衰老相关β-半乳糖苷酶（SA-β-gal）活性染色评估HCC细胞的肝功能和细胞衰老情况。进行RNA测序（RNA-Seq）分析，以鉴定用P-T3H2处理的Huh7细胞中差异表达的基因。在体外和体内评估P-T3H2对HCC恶性程度的影响。
在人和小鼠HCC组织中，TRIB3与HNF4α均呈负相关。给予P-T3H2可显著抑制HCC细胞的恶性程度。此外，P-T3H2可稳定HNF4α蛋白，并促进HCC细胞肝功能的恢复和细胞衰老。RNA-Seq分析表明，P-T3H2可增强HCC中HNF4α的转录活性。此外，P-T3H2可有效抑制小鼠HCC的发生和进展。
P-T3H2通过稳定HNF4α蛋白抑制HCC进展，可能是HCC临床治疗中有前景的候选治疗药物。
