Sun J, Lun Y Z, Liu B, Dong W
Key Laboratory of Screening and Control of Infectious Diseases, Fujian Provincial University, Quanzhou Medical College, Quanzhou, China.
Department of Laboratory Medicine, Putian University, Putian, China.
Bull Exp Biol Med. 2024 Dec;178(2):237-243. doi: 10.1007/s10517-025-06314-0. Epub 2025 Jan 7.
The mechanism of Hespintor (a protein of serpin family) inhibitory action on the growth of inoculated hepatocellular carcinoma was studied in a model of human hepatoma in nude mice by using on long-noncoding RNA (lncRNA) sequencing. Two days after tumor transplantation, Hespintor or normal saline was injected into the caudal vein at a dose of 15 μg/kg (2 times a week over 4 weeks). The tumors were isolated in 4 weeks after subcutaneous injection of human hepatoma MHCC97-H cells. In Hespintor and control groups, the complementary DNA libraries of tumor tissues were established, and transcriptome sequencing was performed. Based on RNA-sequencing data, the differentially expressing lncRNA genes (DEGs lncRNA) were obtained, and functional enrichment and interaction analyses were performed to find the regulatory gene sets. Then, the network module division method was employed to identify the key genes of the Hespintor action, as well as to build the regulatory network and critical pathways associated with the key genes with validation of the results by Western blotting. The target gene sets regulated by DEGs lncRNA were mainly enriched in cell behavior, transcriptional regulation, and cell cycle. The PI3K/Akt signaling pathway related to the revealed gene sets plays a leading role in the antitumor action of Hespintor, targeted by this serpin to down-regulate expression levels of the cell cycle regulatory proteins Cyclin D1, P-Rb, CDK4, and CDK6, thereby arresting the cell cycle in G1/S phase.
通过长链非编码RNA（lncRNA）测序，在裸鼠人肝癌模型中研究了丝氨酸蛋白酶抑制剂家族蛋白Hespintor对接种的肝细胞癌生长的抑制作用机制。肿瘤移植两天后，以15μg/kg的剂量经尾静脉注射Hespintor或生理盐水（每周2次，共4周）。皮下注射人肝癌MHCC97-H细胞4周后分离肿瘤。在Hespintor组和对照组中，建立肿瘤组织的互补DNA文库并进行转录组测序。基于RNA测序数据，获得差异表达的lncRNA基因（DEGs lncRNA），并进行功能富集和相互作用分析以找到调控基因集。然后，采用网络模块划分方法鉴定Hespintor作用的关键基因，构建与关键基因相关的调控网络和关键途径，并通过蛋白质印迹法验证结果。DEGs lncRNA调控的靶基因集主要富集于细胞行为、转录调控和细胞周期。与所揭示的基因集相关的PI3K/Akt信号通路在Hespintor的抗肿瘤作用中起主导作用，这种丝氨酸蛋白酶抑制剂靶向该通路以下调细胞周期调节蛋白Cyclin D1、P-Rb、CDK4和CDK6的表达水平，从而使细胞周期停滞在G1/S期。
