METHODS: SerpinB9 expression in hepatoblastoma cell lines and macrophages co-cultured with each other or stimulated by anticancer agents was examined using RT-qPCR and western blotting. Immunohistochemistry for SerpinB9 in hepatoblastoma specimens was performed. Single-cell RNA-sequence data for hepatoblastoma from an online database were analyzed to investigate which types of cells express SerpinB9.
RESULTS: HepG2, a hepatoblastoma cell line, exhibited increased expression of SerpinB9 when indirectly co-cultured with macrophages. Immunohistochemistry for the specimens demonstrated that serpinB9 is positive not in hepatoblastoma cells but in macrophages. Single-cell RNA sequence analysis in tissues from hepatoblastoma patients showed that macrophages expressed SerpinB9 more than tumor cells did. Co-culture of macrophages with hepatoblastoma cell lines led to the enhanced expression of SerpinB9 in both macrophages and cell lines. Anticancer agents induced an elevation of SerpinB9 in hepatoblastomas cell lines.
CONCLUSIONS: In hepatoblastoma, SerpinB9 is thought to be more highly expressed in macrophages and enhanced by interaction with hepatoblastoma cell.
方法:使用RT-qPCR和western印迹检测了彼此共培养或由抗癌剂刺激的肝母细胞瘤细胞系和巨噬细胞中SerpinB9的表达。对肝母细胞瘤标本中的SerpinB9进行免疫组织化学。分析了来自在线数据库的肝母细胞瘤的单细胞RNA序列数据,以研究哪些类型的细胞表达SerpinB9。
结果:HepG2,一种肝母细胞瘤细胞系,当与巨噬细胞间接共培养时,SerpinB9的表达增加。标本的免疫组织化学表明,serpinB9不是在肝母细胞瘤细胞中而是在巨噬细胞中呈阳性。肝母细胞瘤患者组织中的单细胞RNA序列分析表明,巨噬细胞比肿瘤细胞更表达SerpinB9。巨噬细胞与肝母细胞瘤细胞系的共培养导致SerpinB9在巨噬细胞和细胞系中的表达增强。抗癌剂诱导肝母细胞瘤细胞系中SerpinB9的升高。
结论:在肝母细胞瘤中,SerpinB9被认为在巨噬细胞中更高表达,并通过与肝母细胞瘤细胞的相互作用而增强。