Transcriptome analysis of immune-inflammatory regulation in Tremella fuciformis-derived polysaccharide reeducated B16 cells: A subcutaneous model.

BACKGROUND: Circulating cancer cells have characteristics of tumor self-targeting. Modified circulating tumor cells may serve as tumor-targeted cellular drugs. Tremella fuciformis-derived polysaccharide (TFP) is related to immune regulation and tumor inhibition, so could B16 cells reeducated by TFP be an effective anti-tumor drug? OBJECTIVES: To evaluate the intrinsic therapeutic potential of B16 cells exposed to TFP and clarify the therapeutic molecules or pathways altered by this process. MATERIAL AND METHODS: RNA-seq technology was used to study the effect of TFP-reeducated B16 cells on the immune and inflammatory system by placing the allograft subcutaneously in C57BL/6 mice. RESULTS: Tremella fuciformis-derived polysaccharide-reeducated B16 cells recruited leukocytes, neutrophils, dendritic cells (DCs), and mast cells into the subcutaneous region and promoted the infiltration of several cytokines such as tumor necrosis factor alpha (TNF-α), interleukin 6 (IL-6), interleukin 1ß (IL-1ß), and interleukin 1 (IL-1). Tumor necrosis factor alpha also activated Th17 lymphocytes to secrete interleukin 17 (IL-17) and interferon gamma (IFN-γ). The co-expression of IFN-γ and IL-17 was favorable for tumor immunity to shrink tumors. In short, TFP-reeducated B16 cells activated the innate and adaptive immune responses, especially Th17 cell differentiation and IFN-γ production, as well as the TNF-α signaling pathway, which re-regulated the inflammatory and immune systems. CONCLUSION: B16 cells subcutaneously exposed to TFP in mice induced an immune and inflammatory response to inhibit tumors. The study of the function of TFP-reeducated B16 cells to improve cancer immunotherapy may be of particular research interest. This approach could be an alternative and more efficient strategy to deliver cytokines and open up new possibilities for long-lasting, multi-level tumor control.

Overcharged lipid metabolism in mechanisms of antitumor by Tremella fuciformis­derived polysaccharide.

Tremella fuciformis­derived polysaccharide (TFP) is a natural macromolecular compound that is well known for whitening skin, as well as for its ability to regulate lipids and immunity. However, its mechanism of action is not clear. In the present study, B16 cells treated with TFP were inoculated subcutaneously in the right flank of C57BL/6 mice to explore the effect of TFP on melanoma in vivo. Western blotting, immunohistochemistry, immunofluorescence staining and transcription analysis of tumors were utilized to assess the expression of key molecules in production of melanin, lipid metabolism and immunity. It was found that TFP promoted B16 cell apoptosis and induced G2/M cell cycle arrest, which was associated with activation of cell cycle­related pathways. TFP induced the expression of glucose transporter type 4 and CD36, thus resulting in an increase in the uptake of lipids, which markedly suppressed sterol regulatory element­binding transcription factor 1 and phosphorylated­AMP­activated protein kinase expression; increased the number of lipids in the cell membrane, endoplasmic reticulum and nucleus; and induced the RNA expression of molecules related to lipid metabolism, as revealed by RNA­sequencing in vivo. Increased lipid binding, upregulated lipid storage, and elevated triglyceride and lipid catabolism resulted in disruption of cell volume homeostasis and activated innate immune response, thus inhibiting melanoma development and progression. These data revealed a novel molecular mechanism involved in the antitumor effect of TFP via lipid metabolism.

Sulfhydryl functionalized hyaluronic acid hydrogels attenuate cyclophosphamide-induced bladder injury.

Clinical management of cyclophosphamide (CYP) results in numerous side effects including hemorrhagic cystitis (HC), which is characterized by inflammation and oxidative stress damage. Intravesical hyaluronic acid (HA) supplementation, a therapeutic method to restore barrier function of bladder, avoid the stimulation of metabolic toxicants on bladder and reduce inflammatory response, has shown good results in acute or chronic bladder diseases. However, there are unmet medical needs for the treatment of HC to temporarily restore bladder barrier and reduce inflammation. Herein, sulfhydryl functionalized HA (HA-SH) and dimethyl sulfoxide (DMSO) were used to prepared a hydrogel system for optimizing the treatment of HC. We systematically evaluated the physicochemical of hydrogels and their roles in a rat model of CYP-induced HC. The prepared hydrogels exhibited outstanding gel forming properties, injectability, and biosafety. Swelling and retention studies showed that hydrogels were stable and could prolong the residence time of HA in the bladder. Histopathology and vascular permeability studies indicated that the hydrogels significantly attenuated bladder injury caused by CYP administration. Moreover, the hydrogels also showed excellent anti-inflammation and anti-oxidation properties. In conclusion, these data suggest that intravesical instillation of HA-SH/DMSO hydrogels reduces CYP-induced bladder toxicity and this work provides a new strategy for the prevention and early treatment of HC.