Sha Meng, Zhang Jiang, Liu Jin-Kai, Qu Xiao-Ye, Shen Chuan, Tong Ying, Cao Jie
Department of Liver Surgery, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, 160 Pujian Road, Shanghai, 200127, China.
Cancer Cell Int. 2025 Jun 7;25(1):205. doi: 10.1186/s12935-025-03841-8.
Liver cancer stem cells (LCSCs) are critical drivers of metastasis and chemoresistance in hepatocellular carcinoma (HCC). Proline-rich tyrosine kinase 2 (Pyk2) has been implicated in tumor progression, but its role in LCSC stemness and HCC malignancy remains unclear. This study explores the effects of Pyk2 and its regulation by miR-23b-3p on LCSC function and HCC progression.
LCSCs were enriched from HepG2 and HCCLM3 cell lines, and Pyk2 knockdown was induced through siRNA transfection, with or without miR-23b-3p inhibitor co-transfection. We assessed cell proliferation, sphere formation, migration, invasion, and chemosensitivity. Stemness markers (Nanog, Oct4, Sox2, KLF4, and Bmi1) and Pyk2/Src signaling were analyzed via RT-qPCR, Western blotting, and immunohistochemistry. In vivo, tumor growth and Pyk2/Src expressions were evaluated in a BALB/c mouse xenograft model.
Pyk2 expression was significantly elevated in the identified LCSCs compared to the parental HCCs. Pyk2 knockdown significantly suppressed the LCSCs proliferation, sphere formation, migration, invasion, and enhanced chemosensitivity. The expression of stemness markers and miR-23b-3p was significantly inhibited in HCCLM3-LCSC cells. miR-23b-3p inhibition restored Pyk2 level and Src phosphorylation, reversing the suppressive effects of Pyk2 knockdown. In BALB/c mice, tumor volume, weight, and Pyk2/Src expressions were significantly elevated in HCCLM3-LCSC and HCCLM3-LCSC+miR-23b-3p inhibitor groups comparing to HCCLM3/HCCLM3-LCSC groups, whereas were even heightened in the HCCLM3-LCSC + miR-23b-3p inhibitor group.
Inhibiting Pyk2/Src expression by miR-23b-3p suppressed LCSCs function and aggravated HCC progression.
Not applicable.
肝癌干细胞（LCSCs）是肝细胞癌（HCC）转移和化疗耐药的关键驱动因素。富含脯氨酸的酪氨酸激酶2（Pyk2）与肿瘤进展有关，但其在LCSC干性和HCC恶性肿瘤中的作用仍不清楚。本研究探讨Pyk2及其受miR-23b-3p调控对LCSC功能和HCC进展的影响。
从HepG2和HCCLM3细胞系中富集LCSCs，通过siRNA转染诱导Pyk2基因敲低，同时或不同时共转染miR-23b-3p抑制剂。我们评估了细胞增殖、成球、迁移、侵袭和化疗敏感性。通过RT-qPCR、蛋白质印迹和免疫组织化学分析干性标志物（Nanog、Oct4、Sox2、KLF4和Bmi1）以及Pyk2/Src信号通路。在体内，在BALB/c小鼠异种移植模型中评估肿瘤生长和Pyk2/Src表达。
与亲本HCC相比，在鉴定出的LCSCs中Pyk2表达显著升高。Pyk2基因敲低显著抑制LCSCs增殖、成球、迁移、侵袭，并增强化疗敏感性。在HCCLM3-LCSC细胞中，干性标志物和miR-23b-3p的表达显著受到抑制。miR-23b-3p抑制恢复了Pyk2水平和Src磷酸化，逆转了Pyk2基因敲低的抑制作用。在BALB/c小鼠中，与HCCLM3/HCCLM3-LCSC组相比，HCCLM3-LCSC和HCCLM3-LCSC+miR-23b-3p抑制剂组的肿瘤体积、重量和Pyk2/Src表达显著升高，而在HCCLM3-LCSC+miR-23b-3p抑制剂组中甚至更高涨。
miR-23b-3p抑制Pyk2/Src表达可抑制LCSCs功能并加剧HCC进展。
不适用。
