Li Jian, Qin Yuanhua, Yang Yingying, Chen Hang, Li Mengjuan, Liu Yadi, Liu Bingjie, Shang Jingli, Zhang Yu, Han Tao, Hu Yuhan, Ren Feng
Department of Pathology, the First Affiliated Hospital of Xinxiang Medical University, Xinxiang, 453003, China.
School of Basic Medical Sciences, Xinxiang Medical University, Xinxiang, 453003, China.
Biol Proced Online. 2025 Jun 17;27(1):22. doi: 10.1186/s12575-025-00282-5.
Hepatocellular carcinoma (HCC) ranks as the third most common malignant tumor globally. Although hyperthermia has shown promise as a non-invasive treatment for tumors, its specific mechanisms and impact on HCC remain underexplored. This study aims to investigate the effects of hyperthermia on HCC proliferation and apoptosis and to elucidate the role of the Hippo signaling pathway in these processes. Huh7 and HepG2 HCC cells were cultured at various temperatures (37 °C, 40 °C, 43 °C, and 46 °C), with 37 °C as the control. Cell proliferation, apoptosis, and cell cycle distribution were assessed via CCK-8 assays and flow cytometry. A subcutaneous xenograft model in nude mice, treated with indocyanine green (ICG) and near-infrared (NIR) irradiation to achieve local hyperthermia, was used to evaluate in vivo tumor growth. RNA-seq and KEGG pathway analyses identified differentially expressed genes, and Western blotting and immunofluorescence were used to confirm the involvement of the Hippo signaling pathway. Hyperthermia at 43 °C significantly inhibited Huh7 and HepG2 cell proliferation and induced apoptosis, accompanied by cell cycle arrest. In vivo, local hyperthermia reduced tumor volume and weight in the ICG + NIR-treated group. RNA-seq and KEGG analyses revealed that the Hippo signaling pathway was activated under hyperthermic conditions, with YAP expression and nuclear translocation markedly downregulated. Further experiments showed that YAP overexpression mitigated hyperthermia-induced effects on cell proliferation and apoptosis, underscoring the role of the Hippo pathway. These findings demonstrate that hyperthermia inhibits HCC growth by regulating the Hippo signaling pathway, reducing cell proliferation, and promoting apoptosis. This study highlights the potential of hyperthermia as an effective therapeutic approach for HCC, with implications for developing targeted hyperthermic therapies.
肝细胞癌（HCC）是全球第三大常见恶性肿瘤。尽管热疗已显示出作为肿瘤非侵入性治疗方法的潜力，但其具体机制以及对HCC的影响仍未得到充分研究。本研究旨在探讨热疗对HCC增殖和凋亡的影响，并阐明Hippo信号通路在这些过程中的作用。将Huh7和HepG2肝癌细胞在不同温度（37℃、40℃、43℃和46℃）下培养，以37℃作为对照。通过CCK-8测定和流式细胞术评估细胞增殖、凋亡和细胞周期分布。使用经吲哚菁绿（ICG）和近红外（NIR）照射以实现局部热疗的裸鼠皮下异种移植模型来评估体内肿瘤生长。RNA测序和KEGG通路分析确定了差异表达基因，并使用蛋白质印迹和免疫荧光来确认Hippo信号通路的参与情况。43℃的热疗显著抑制Huh7和HepG2细胞增殖并诱导凋亡，同时伴有细胞周期停滞。在体内，局部热疗降低了ICG + NIR治疗组的肿瘤体积和重量。RNA测序和KEGG分析显示，热疗条件下Hippo信号通路被激活，YAP表达和核转位明显下调。进一步的实验表明，YAP过表达减轻了热疗对细胞增殖和凋亡的影响，突出了Hippo通路的作用。这些发现表明，热疗通过调节Hippo信号通路、减少细胞增殖和促进凋亡来抑制HCC生长。本研究强调了热疗作为HCC有效治疗方法的潜力，对开发靶向热疗具有启示意义。
