Disruption of crosstalk between LX-2 and liver cancer stem-like cells from MHCC97H cells by DFOG via inhibiting FOXM1
Disruption of crosstalk between LX-2 and liver cancer stem-like cells from MHCC97H cells by DFOG via inhibiting FOXM1
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DOI：
10.1093/abbs/gmz129
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发表时间：
2019-12-01
影响因子：
3.7
通讯作者：
Li, Xiang
中科院分区：
文献类型：
--
作者：
Chen, A.;Xu, Chang;Li, Xiang
肝星状细胞 (HSC) 系 LX-2 被肝癌干样细胞 (LCSLC) 激活，并产生构成大部分肝细胞癌 (HCC) 微环境的各种细胞因子。新的染料木黄酮衍生物 7-二氟甲氧基-5,4'-二正辛基染料木黄酮 (DFOG) 通过控制叉头盒 M1 (FOXM1) 在多种恶性肿瘤中显示出抗癌作用。在本研究中，我们旨在评估 DFOG 是否会破坏人类 HSC LX-2 细胞和 LCSLC 之间的串扰。获得了不同世代的 MHCC97H 衍生球体，第二代被认为是 LCSLC，其表现出增强的自我更新能力和升高的 CD133、CD44 和 EpCAM 蛋白表达水平，以及致瘤性，如体外集落形成测定和体内致瘤性测定所揭示的。使用 Transwell 系统将 LX-2 和 MHCC97H 细胞与/不与 DFOG（分别为 1、5 和 10 muM）共培养。 FOXM1 过表达和/或敲低用于机制研究。我们的结果表明，Co-CM 促进 LX-2 细胞转化为肝癌相关 HSC。同时，Co-CM刺激后，LX-2细胞和上清液中FOXM1上调，肝细胞生长因子（HGF）水平增加。 MHCC97H 细胞的球体和集落形成能力以及 CD133、CD44 和 EpCAM 的蛋白水平也显着升高。 DFOG剂量依赖性地抑制上述效应，类似于LX-2细胞中FOXM1敲低。 FOXM1 过表达逆转了 DFOG 或 FOXM1 敲低或两者对 LCSLC/LX-2 共培养的 MHCC97H 细胞中 LX-2 细胞激活和 LCSLC 特征诱导的抑制作用。这项研究表明，DFOG 通过下调 FOXM1 表达和减少 HSC 中 HGF 分泌来破坏 HSC 和 LCSLC 之间的串扰，从而抑制 LCSLC 特征。
Hepatic stellate cell (HSC) line LX-2 is activated by liver cancer stem-like cells (LCSLCs) and produces various cytokines that make up most of the hepatocellular carcinoma (HCC) microenvironment. The new genistein derivative, 7-difluoromethoxyl-5,4'-di-n-octylgenistein (DFOG), shows anticancer effects in multiple malignancies by controlling forkhead box M1 (FOXM1). In this study, we aimed to assess whether DFOG disrupts the crosstalk between human HSC LX-2 cells and LCSLCs. Distinct generations of MHCC97H-derived spheres were obtained with the second generation considered as LCSLCs which displayed enhanced self-renewal ability and elevated expression levels of CD133, CD44, and EpCAM proteins, as well as tumorigenicity, as revealed by colony formation assay in vitro and tumorigenicity assay in vivo. LX-2 and MHCC97H cells were co-cultured with/without DFOG (1, 5, and 10 mu M, respectively) using the transwell system. FOXM1 overexpression and/or knockdown were employed for mechanistic investigations. Our results suggested that Co-CM promoted LX-2 cell transformation into liver cancer-associated HSCs. Meanwhile, FOXM1 was up-regulated and the level of hepatocyte growth factor (HGF) was increased in LX-2 cells and in the supernatant after Co-CM stimulation. Sphere and colony formation abilities in MHCC97H cells, and protein levels of CD133, CD44, and EpCAM, were also markedly elevated. DFOG dose-dependently inhibited the above effects, similar to FOXM1 knockdown in LX-2 cells. FOXM1 overexpression reversed the inhibitory effects of DFOG or FOXM1 knockdown or both on LX-2 cell activation and LCSLC feature induction in MHCC97H cells by LCSLC/LX-2 co-culture. This study demonstrated that DFOG disrupts the crosstalk between HSCs and LCSLCs to suppress LCSLC features via down-regulating FOXM1 expression and reducing HGF secretion in HSCs.