目的 探索巨噬细胞在肝癌炎症微环境中通过炎症趋化因子(C-X-C基序)配体1(CXCL1)调控肝癌细胞系Huh7进展.方法 筛选肿瘤临床数据库TISIDB,分析CX-CL1与肝癌患者预后的关系.免疫组化、免疫荧光染色验证肿瘤巨噬细胞与CXCL1的关系.通过佛波酯(PMA)诱导人单核细胞(THP-1)获得未分化的巨噬细胞(M0),建立与肝癌细胞共培养模型,实时荧光定量PCR法检测巨噬细胞表型的改变;细胞克隆形成实验、台盼蓝细胞活率检测肝癌细胞的增殖能力;肝癌细胞的迁移及侵袭能力使用Transwell实验检测;肝癌细胞Huh7中E-cadherin、N-cadherin、Vimentin的蛋白表达水平使用蛋白印迹法检测;酶联免疫吸附实验(ELISA法)检测细胞培养液中目标蛋白的表达.结果 CXCL1高表达与肝癌患者不良预后有关,并且高表达CX-CL1的肝癌组织具有更多的巨噬细胞富集.在巨噬细胞-肝癌细胞共培养模型中,两种细胞CXCL1的表达水平及共培养液中CXCL1浓度均增加.巨噬细胞M2型标志物CD163、CD206 mRNA表达水平升高.肝癌细胞Huh7的增殖、迁移及侵袭能力增强.结论 巨噬细胞-肝癌细胞共培养增加肿瘤相关巨噬细胞CXCL1表达,促进肝癌细胞系Huh7进展.

Abstract Inflammation is a core mechanism for oncogenesis. Chemokines act as important inflammation mediators in chronic inflammation and the tumor inflammatory response. However, limited information is known about chemokines in hepatocellular carcinoma (HCC), a disease that is almost entirely derived from chronic liver inflammation. Here, we explored the protumor effects of CXCL1, a commonly elevated inflammatory chemokine in cirrhosis, in HCC. This protumor feature was confirmed in clinical samples of human HCC. CXCL1 enhances tumorigenesis in the hepatic inflammatory microenvironment directly through tumor cells and indirectly through recruitment of macrophages. Increasing the number of macrophages in the tumor microenvironment (TME) promoted tumor cell Epithelial-mesenchymal transition (EMT) capacity and significantly elevated CXCL1 levels in the TME partly through NF-κB/IL-1𝛽 activation. To investigate the potential therapeutic value of CXCL1 in HCC with inflammatory background, blocking CXCL1 and blocking CXCL1 combined with the chemotherapy agent doxorubicin (DOX), which aimes to reshape the TME, were administered. It has been shown that blocking CXCL1-CXCR2 inhibits tumor progression and reduces macrophage recruitment in the TME. The combination regimen has been shown to have a synchronous effect in HCC by reducing pro-tumor macrophages in the TME and suppressing tumor cell progression. This provides insight into therapeutic strategies for treating HCC patients with high CXCL1 expression.

Inflammation is a core mechanism for oncogenesis. Chemokines act as important mediators of chronic inflammation and the tumour inflammatory response. However, there is limited information on chemokines in hepatocellular carcinoma (HCC), a disease for which almost all cases are derived from chronic liver inflammation. Here, we explored the protumor effects of CXCL1, a commonly elevated inflammatory chemokine in cirrhosis, in HCC. The protumor role was confirmed in clinical samples from HCC patients. CXCL1 enhanced tumorigenesis in the hepatic inflammatory microenvironment directly by acting on tumour cells and indirectly through promoting the recruitment of macrophages. The increase in the number of macrophages in the tumour microenvironment (TME) promoted tumour cell epithelial-mesenchymal transition (EMT) and significantly increased CXCL1 levels in the TME partly through NF-κB/IL-1 β activation. To investigate the potential therapeutic value of CXCL1 in HCC with an inflammatory background, an antibody blocking CXCL1 was used alone or combined with the chemotherapy agent doxorubicin (DOX), with the goal of reshaping the TME. It has been shown that blocking CXCL1-CXCR2 inhibits tumour progression and reduces macrophage recruitment in the TME. The combination regimen has been shown to synergistically reduce the number of pro-tumour macrophages in the TME and suppress tumour progression. This provides insight into therapeutic strategies for treating HCC patients with high CXCL1 expression.

Tumor heterogeneity dominates tumor biological behavior and shapes the tumor microenvironment. However, the mechanisms of tumor genetic features modulate immunity response were not clearly clarified. Tumor as-sociated macrophages (TAMs) exert distinct immune functions in the progression of hepatocellular carcinoma (HCC) based on the inducible phenotype. FOXO family members sense changes in the extracellular or intracel-lular environment by activating a series of signaling pathways. FOXO1, a transcription factor that a common suppressor in hepatocellular carcinoma, correlated with a better tumor biological behavior in HCC through shap-ing macrophages anti-tumour response. Here, we found that human HCC tissue microarray (TMA) slides were employed to showed tumor derived FOXO1 negatively related with distribution of protumour macrophages. This phenomenon was confirmed in mouse xenograft model and in vitro . HCC-derived FOXO1 inhibits tumorigenesis not only by targeting tumor cells but also by synchronizing with re-educated macrophages. These effects may be partially dependent on FOXO1 transcriptionally modulates IRF-1/nitrio oxide (NO) axis in exerting effects in macrophages and decreasing IL-6 releasing from macrophages in tumor microenvironment indirectly. This feed-back suppressed the progression of HCC by inactivation of IL-6/STAT3 in HCC. It implicates the potential role of FOXO1 in the therapeutic effects for modulating immune response by targeting macrophages.

目的 研究microRNA-200a(miR200a)影响肝细胞癌的进展及调控索拉非尼治疗效果.方法 构建人肝癌细胞miR200a稳转细胞株:miR200a过表达、对照组(Control)、miR200a敲低.采用qRT-PCR法检测各转染细胞株miRNA表达量,MTT的方法检测细胞活力,Western blot分析各目标蛋白表达量;Transwell法检测肝癌细胞迁移、侵袭能力.转染细胞株添加索拉非尼,复测MTT及Western blot,验证miR200a调控索拉非尼对细胞的作用和相应蛋白表达量变化,SPF级BALB/c-nu雄鼠18只,随机分为3组,每组6只.每组皮下种植miR200a过表达、Control、miR200a敲低细胞,待成瘤后每组裸鼠每天注射索拉非尼,连续14 d,处死裸鼠,取出瘤体,计算肿瘤体积,免疫组化验证瘤体目标蛋白表达及定位.结果 上调miR200a不仅可以抑制人肝癌细胞的细胞活力及迁移侵袭能力,还可以降低SIRT1的表达.并且上调miR200a明显减小裸鼠的肿瘤体积.结论 上调miR200a可以负性调控人肝癌细胞的增殖和迁移能力,并增加索拉非尼的药物疗效,这种作用可能是通过调控SIRT1表达来完成.

Hepatocellular carcinoma (HCC) is one of the most common tumours worldwide, and identifying markers related to HCC is an important area of research. As a microRNA (miRNA), miRNA125b (miR-125b) plays an important role in the prediction and prognosis of HCC. In the past 10 years, with increasing research on miR-125b and HCC, the molecular mechanism of its relationship with the development of HCC has been elucidated. MiR-125b inhibits the development of HCC and is highly accurate in predicting HCC and is therefore a valuable predictive marker of HCC. This article summarizes the clinical application of miR-125b in HCC and the potential mechanism of its involvement in the progression of HCC.