Batsaikhan Bat-Erdene, Yoshikawa Kozo, Kurita Nobuhiro, Iwata Takashi, Takasu Chie, Kashihara Hideya, Shimada Mitsuo
Department of Surgery, Institute of Health Biosciences, The University of Tokushima, Tokushima, Japan.
Department of Surgery, Institute of Health Biosciences, The University of Tokushima, Tokushima, Japan
Anticancer Res. 2014 Nov;34(11):6339-44.
Backround: Most solid cancers including colon cancer are believed to be initiated from and maintained by cancer stem cells (CSCs), that are responsible for treatment resistance, resulting in tumor relapse. The aim of this study was to clarify the possible role of the Sonic Hedgehog (Shh) signaling pathway in the regulation of cancer stem cells.
The HCT-116 cell line was cultured with fetal bovine serum in RPMI-1640 medium and its sphere was grown in serum-free non-adherent culture. Gene expressions were analyzed by quantitative real-time polymerase chain reaction (qRT-PCR) from cells treated with and without cyclopamine.
HCT-116 sphere-derived cells grown in serum-free, non-adherent culture, showed significantly increased expression of stem cell markers, Shh downstream genes and epithelial-mesenchymal transition (EMT) markers compared to parental cells grown in conventional culture. The expression of stemness markers, Shh downstream genes and EMT markers were higher in cancer spheres than the parental cell line and down-regulated by cyclopamine treatment in a dose-dependent manner.
Overall, these findings show that cyclopamine treatment could down-regulate the expression of stemness markers, shh downstream genes and EMT markers on HCT-116 spheres.
背景：包括结肠癌在内的大多数实体癌被认为是由癌症干细胞（CSCs）启动并维持的，这些癌症干细胞导致治疗耐药，进而引发肿瘤复发。本研究的目的是阐明 Sonic Hedgehog（Shh）信号通路在癌症干细胞调控中的可能作用。
HCT-116 细胞系在含有胎牛血清的 RPMI-1640 培养基中培养，其球体在无血清非贴壁培养中生长。通过定量实时聚合酶链反应（qRT-PCR）分析用环杷明处理和未处理的细胞的基因表达。
与在传统培养中生长的亲代细胞相比，在无血清非贴壁培养中生长的 HCT-116 球体来源细胞显示干细胞标志物、Shh 下游基因和上皮-间质转化（EMT）标志物的表达显著增加。癌症球体中干性标志物、Shh 下游基因和 EMT 标志物的表达高于亲代细胞系，并通过环杷明处理以剂量依赖方式下调。
总体而言，这些发现表明环杷明处理可下调 HCT-116 球体上干性标志物、shh 下游基因和 EMT 标志物的表达。
