Sulfated Hyaluronan Binds to Heparanase and Blocks Its Enzymatic and Cellular Actions in Carcinoma Cells

We examined whether sulfated hyaluronan exerts inhibitory effects on enzymatic and biological actions of heparanase, a sole endo-beta-glucuronidase implicated in cancer malignancy and inflammation. Degradation of heparan sulfate by human and mouse heparanase was inhibited by sulfated hyaluronan. In particular, high-sulfated hyaluronan modified with approximately 2.5 sulfate groups per disaccharide unit effectively inhibited the enzymatic activity at a lower concentration than heparin. Human and mouse heparanase bound to immobilized sulfated hyaluronan. Invasion of heparanase-positive colon-26 cells and 4T1 cells under 3D culture conditions was significantly suppressed in the presence of high-sulfated hyaluronan. Heparanase-induced release of CCL2 from colon-26 cells was suppressed in the presence of sulfated hyaluronan via blocking of cell surface binding and subsequent intracellular NF-κB-dependent signaling. The inhibitory effect of sulfated hyaluronan is likely due to competitive binding to the heparanase molecule, which antagonizes the heparanase-substrate interaction. Fragment molecular orbital calculation revealed a strong binding of sulfated hyaluronan tetrasaccharide to the heparanase molecule based on electrostatic interactions, particularly characterized by interactions of (−1)- and (−2)-positioned sulfated sugar residues with basic amino acid residues composing the heparin-binding domain-1 of heparanase. These results propose a relevance for sulfated hyaluronan in the blocking of heparanase-mediated enzymatic and cellular actions.     

RON蛋白与CXC趋化因子受体4蛋白的表达与去势抵抗型前列腺癌患者阿比特龙耐药的相关性

目的：研究受体酪氨酸激酶（RON）蛋白与CXC趋化因子受体4（CXCR4）蛋白的表达与去势抵抗型前列腺癌（CRPC）患者阿比特龙耐药的相关性。方法：选取2017年1月至2020年2月我院收治的127例接受阿比特龙治疗的CRPC患者,根据是否耐药分为观察组（n=32,阿比特龙耐药患者）、对照组（n=95,缓解患者）。采用免疫组化与蛋白免疫印迹检测比较两组RON、CXCR4蛋白表达,采用Logistic回归分析进行RON、CXCR4蛋白与耐药的单因素、多因素分析,采用受试者工作特征曲线（ROC）及ROC下面积（AUC）分析RON、CXCR4蛋白预测耐药的价值,并在阿比特龙耐药细胞株中加入RON、CXCR4抑制剂,观察两者对阿比特龙耐药细胞凋亡指标［半胱氨酸蛋白酶（caspase）-3、caspase-9、细胞凋亡率］的影响。结果：免疫组化显示,观察组RON阳性表达率（71.88%,23/32）较对照组（27.37%,26/95）高;观察组CXCR4阳性表达率（65.63%,21/32）较对照组（12.63%,12/95）高;蛋白免疫印迹检测显示,观察组RON、CXCR4蛋白较对照组高（P<0.05）;RON、CXCR4蛋白与耐药均呈正相关（P<0.05）;加入RON、CXCR4抑制剂后,RON、CXCR4表达被成功抑制,且caspase-3、caspase-9、细胞凋亡率均高于阿比特龙耐药细胞株（P<0.05）;Transwell实验检测细胞迁移及侵袭显示,抑制RON、CXCR4表达,细胞迁移及侵袭细胞数目均显著降低（P<0.05）;RON蛋白预测阿比特龙耐药的AUC为0.789,截断值＞4.11,敏感度为84.37%,特异度为61.05%（P<0.05）;CXCR4蛋白预测阿比特龙耐药的AUC为0.825,截断值＞3.42,敏感度为75.00%,特异度为80.00%（P<0.05）;RON+CXCR4蛋白预测阿比特龙耐药的AUC为0.884（95%CI：0.815～0.934）,敏感度为87.50%,特异度为83.16%（P<0.05）。 结论：CRPC患者RON、CXCR4蛋白表达显著增加,与患者阿比特龙耐药密切相关,有望成为预测耐药的标志物,抑制RON、CXCR4蛋白表达,可促进CRPC阿比特龙耐药细胞的凋亡。     

CXC Chemokine Signaling in Progression of Epithelial Ovarian Cancer: Theranostic Perspectives

Patients with epithelial ovarian cancer (EOC) are often diagnosed at an advanced stage due to nonspecific symptoms and ineffective screening approaches. Although chemotherapy has been available and widely used for the treatment of advanced EOC, the overall prognosis remains dismal. As part of the intrinsic defense mechanisms against cancer development and progression, immune cells are recruited into the tumor microenvironment (TME), and this process is directed by the interactions between different chemokines and their receptors. In this review, the functional significance of CXC chemokine ligands/chemokine receptors (CXCL/CXCR) and their roles in modulating EOC progression are summarized. The status and prospects of CXCR/CXCL-based theranostic strategies in EOC management are also discussed.     

The Role of Chemokines in the Development of Gastric Cancer—Diagnostic and Therapeutic Implications

Gastric cancer (GC) is the fifth most common cancer worldwide and the second leading cause of cancer-related death. GC is usually diagnosed at an advanced stage due to late presentation of symptoms. Therefore, there is a need for establishing more sensitive and specific markers useful in early detection of the disease when a cancer is asymptomatic to improve the diagnostic and clinical decision-making process. Some researchers suggest that chemokines and their specific receptors play an important role in GC initiation and progression via promotion of angiogenesis, tumor transformation, invasion, survival and metastasis as well as protection from host response and inter-cell communication. Chemokines are small proteins produced by various cells such as endothelial cells, fibroblasts, leukocytes, and epithelial and tumor cells. According to our knowledge, the significance of chemokines and their specific receptors in diagnosing GC and evaluating its progression has not been fully elucidated. The present article offers a review of current knowledge on general characteristics of chemokines, specific receptors and their role in GC pathogenesis as well as their potential usefulness as novel biomarkers for GC.     

Synthesis and application of fluorescein- and biotin-labeled molecular probes for the chemokine receptor CXCR4

The design, synthesis, and bioevaluation of fluorescence- and biotin-labeled CXCR4 antagonists are described. The modification of D-Lys8 at an epsilon-amino group in the peptide antagonist Ac-TZ14011 derived from polyphemusin II had no significant influence on the potent binding of the peptide to the CXCR4 receptor. The application of the labeled peptides in flow cytometry and confocal microscopy studies demonstrated the selectivity of their binding to the CXCR4 receptor, but not to CXCR7, which was recently reported to be another receptor for stromal cell-derived factor 1 (SDF-1)/CXCL12.     

The Effect of Hypoxia on the Expression of CXC Chemokines and CXC Chemokine Receptors—A Review of Literature

Hypoxia is an integral component of the tumor microenvironment. Either as chronic or cycling hypoxia, it exerts a similar effect on cancer processes by activating hypoxia-inducible factor-1 (HIF-1) and nuclear factor (NF-κB), with cycling hypoxia showing a stronger proinflammatory influence. One of the systems affected by hypoxia is the CXC chemokine system. This paper reviews all available information on hypoxia-induced changes in the expression of all CXC chemokines (CXCL1, CXCL2, CXCL3, CXCL4, CXCL5, CXCL6, CXCL7, CXCL8 (IL-8), CXCL9, CXCL10, CXCL11, CXCL12 (SDF-1), CXCL13, CXCL14, CXCL15, CXCL16, CXCL17) as well as CXC chemokine receptors—CXCR1, CXCR2, CXCR3, CXCR4, CXCR5, CXCR6, CXCR7 and CXCR8. First, we present basic information on the effect of these chemoattractant cytokines on cancer processes. We then discuss the effect of hypoxia-induced changes on CXC chemokine expression on the angiogenesis, lymphangiogenesis and recruitment of various cells to the tumor niche, including myeloid-derived suppressor cells (MDSCs), tumor-associated macrophages (TAMs), tumor-associated neutrophils (TANs), regulatory T cells (T_regs) and tumor-infiltrating lymphocytes (TILs). Finally, the review summarizes data on the use of drugs targeting the CXC chemokine system in cancer therapies.     

6Ckine修饰的树突状细胞促进T淋巴细胞增殖和分化的作用

目的研究次级淋巴组织趋化因子(6Ckine)修饰的树突状细胞(DC)对T淋巴细胞增殖和分化的影响。方法用携带人6Ckine基因的重组复制缺陷型腺病毒(Ad-6Ckine)感染人外周血单个核细胞来源的DC,检测Ad-6Ckine-DC对6Ckine的表达及细胞因子分泌的影响,并观察其吞噬功能和表型的变化及对自身T淋巴细胞的趋化作用。用结肠癌LoVo细胞抗原致敏Ad-6Ckine-DC,将该DC与自身T淋巴细胞共同培养,分别用3H掺入法、RT-PCR和ELISA检测Ad-6Ckine-DC对T淋巴细胞增殖和分化的影响。结果在Ad-6Ckine转染后24h内,DC的吞噬功能几乎不受影响。转染的6Ckine基因能在DC中表达,表达的6Ckine能促进其表达CD83和CCR7,上调RANTES的表达。Ad-6Ckine-DC对自身T淋巴细胞有明显的趋化作用,抗原致敏的Ad-6Ckine-DC能显著促进T淋巴细胞的增殖,并增强其表达T-bet和IL-2的能力。结论6Ckine基因的修饰能在一定程度上促进DC的成熟,并募集T淋巴细胞于DC周围,有利于DC向T淋巴细胞传递抗原和第二信息,增强DC促进T淋巴细胞增殖的作用并使其向Th1分化,诱导细胞免疫,将成为制备肿瘤疫苗的一种良好选择。