Pifithrin-μ抑制TSC2突变肝癌细胞株增殖

doi:10.16352/j.issn.1001-6325.2026.08.1059

基础医学与临床 ›› 2026, Vol. 46 ›› Issue (8): 1059-1067.doi: 10.16352/j.issn.1001-6325.2026.08.1059

Pifithrin-μ抑制TSC2突变肝癌细胞株增殖

范文怡¹, 陈瑜炜², 郑翠婷¹, 吕佳蕊³, 王亚南^1*

中国医学科学院北京协和医学院基础医学研究所 1.生理学系;2.生物化学与分子生物学系,北京 100005;
3.中国医科大学附属第一医院器官移植暨肝胆外科,辽宁沈阳 110001

收稿日期:2025-11-11 修回日期:2025-12-30 出版日期:2026-08-05 发布日期:2026-07-22
通讯作者: ^*shlwangyanan@gmail.com
基金资助:
细胞生态海河实验室创新基金(22HHXBSS00012)

Pifithrin-μ inhibits the proliferation of TSC2 mutant hepatocellular carcinoma cells

FAN Wenyi¹, CHEN Yuwei², ZHENG Cuiting¹, LYU Jiarui³, WANG Yanan^1*

1. Department of Physiology; 2. Department of Biochemistry and Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100005;
3. Department of Organ Transplantation and Hepatobiliary Surgery, the First Affiliated Hospital of China Medical University, Shenyang 110001, China

Received:2025-11-11 Revised:2025-12-30 Online:2026-08-05 Published:2026-07-22
Contact: ^*shlwangyanan@gmail.com

摘要/Abstract

摘要： 目的探究Pifithrin-μ(Pifi-μ) 对TSC2突变的肝癌细胞增殖的影响及潜在作用机制。方法采用多种Tsc2突变的小鼠胚胎成纤维细胞(MEFs)及肝癌细胞株,通过CCK8法检测Pifithrin-μ处理后的细胞增殖能力;利用慢病毒感染技术构建TSC2敲低及过表达的稳转肝癌细胞株,Western blot检测TSC2及mTOR通路下游的蛋白质表达,CCK8法检测其对Pifithrin-μ的敏感性;建立裸鼠皮下移植瘤模型进行体内药效评估;采用RNA测序进行差异基因及通路富集分析,并通过Western blot及CCK8法进行验证。结果 Pifithrin-μ以剂量依赖性方式,选择性抑制Tsc2突变的MEFs及肝癌细胞株增殖(P＜0.05);在肝癌细胞中敲低TSC2会增强其对Pifithrin-μ的敏感性,而过表达TSC2则能对Pifithrin-μ产生抵抗(P＜0.05);Pifithrin-μ能够在裸鼠体内抑制Tsc2突变的移植瘤生长(P＜0.05);Pifithrin-μ能选择性下调Tsc2突变细胞中Smad7蛋白质表达,导致p-Smad2/3水平升高,过度激活TGF-β/Smad信号通路(P＜0.05);6种细胞死亡抑制剂分别与Pifithrin-μ联合作用发现活性氧抑制剂及TGF-β/Smad通路抑制剂能够逆转Pifithrin-μ诱导的Tsc2突变细胞死亡(P＜0.05)。结论 Pifithrin-μ能选择性抑制TSC2突变的肝癌细胞株增殖,其作用机制与下调Smad7,破坏TGF-β/Smad信号通路平衡有关。

关键词: TSC2, 肝癌, Pifithrin-μ, TGF-β/Smad信号通路, 细胞增殖

Abstract: Objective To investigate the effects of Pifithrin-μ on the proliferation of TSC2 mutant hepatocellular carcinoma cells. Methods The effect of Pifithrin-μ on cell proliferation was evaluated using CCK8 assay in multiple lines of Tsc2 mutant mouse embryonic fibroblasts (MEFs) and human hepatocellular carcinoma cells. Lentiviral infection was used to construct hepatocellular carcinoma cells with knockdown or overexpression of TSC2. The protein expression of TSC2 and the downstream effectors of the mTOR pathway were detected by Western blot. Cellular sensitivity to Pifithrin-μ was detected by CCK8. The nude mouse subcutaneous transplantation tumor model was established for evaluation of drug efficacy in vivo. RNA sequencing was used for differential gene and pathway enrichment analysis, which was verified by Western blot and CCK8. Results Pifithrin-μ selectively inhibited the proliferation of Tsc2 mutated MEFs and hepatocellular carcinoma cells in a dose-dependent manner(P＜0.05). Hepatocellular carcinoma cells with TSC2 knockdown showed increased sensitivity to Pifithrin-μ, whereas those with TSC2 overexpression were resistant (P＜0.05). Pifithrin-μ significantly inhibited the growth of Tsc2 mutant transplantation tumors in nude mice (P＜0.05). Pifithrin-μ downregulated the expression of Smad7 protein in Tsc2 mutant cells, resulting in elevated levels of p-Smad2/3 and over-activation of the TGF-β/Smad signaling pathway (P＜0.05). The combination of each of the six cell death inhibitors with Pifithrin-μ revealed that reactive oxygen species inhibitor and TGF-β/Smad pathway inhibitor were able to reverse Pifithrin-μ-induced cell death in TSC2 mutant cells (P＜0.05). Conclusions Pifithrin-μ selectively inhibits the proliferation of TSC2 mutant hepatocellular carcinoma cells. This effect is associated with the downregulation of Smad7 and the disruption of the TGF-β/Smad signaling pathway balance.

Key words: TSC2, hepatocellular carcinoma, Pifithrin-μ, TGF-β/Smad signaling pathway, cell proliferation

中图分类号:

R735.7

范文怡, 陈瑜炜, 郑翠婷, 吕佳蕊, 王亚南. Pifithrin-μ抑制TSC2突变肝癌细胞株增殖[J]. 基础医学与临床, 2026, 46(8): 1059-1067.

FAN Wenyi, CHEN Yuwei, ZHENG Cuiting, LYU Jiarui, WANG Yanan. Pifithrin-μ inhibits the proliferation of TSC2 mutant hepatocellular carcinoma cells[J]. Basic & Clinical Medicine, 2026, 46(8): 1059-1067.

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Pifithrin-μ抑制TSC2突变肝癌细胞株增殖

Pifithrin-μ inhibits the proliferation of TSC2 mutant hepatocellular carcinoma cells

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