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This research explored the mechanism of ganoderic acid X(GAX) on human hepatocellular carcinoma cell models(HepG2, HuH6) and nonobese diabetic-severe combined immune deficient(NOD-SCID) mouse subcutaneous tumor models using proteomics, aiming to provide a basis for the clinical application of GAX. CCK-8 assay was employed to evaluate the effect of GAX on the viability of HepG2 and HuH6 cells. EdU assay was used to assess the effect of GAX on cell proliferation. Scratch assay was used to examine the effect of GAX on cell migration ability. Hoechst 33258 staining was used to investigate the effect of GAX on cell apoptosis. Moreover, a NOD-SCID mouse subcutaneous tumor model was established to analyze the tumor volume and weight in control group and GAX low-, medium-, and high-dose groups(5, 10, and 20 mg·kg~(-1)). HE staining was conducted to evaluate the drug toxicity of GAX. Additionally, HepG2 cells in the control group and the GAX high-dose group were subjected to label-free proteomics analysis to identify differential proteins and enrich relevant signaling pathways. CYTO-ID® staining was performed to detect autophagy, and Western blot was conducted to measure the expression levels of relevant proteins. In vitro results demonstrated that GAX dose-depen-dently inhibited proliferation, migration, and induced apoptosis in HepG2 and HuH6 cells. In vivo studies showed that GAX significantly inhibited tumor volume and weight without causing significant damage to major organs(heart, liver, spleen, lung, and kidney) in mice. Label-free proteomics analysis revealed that GAX participated in multiple signaling pathways during the treatment of hepatocellular carcinoma, with a high enrichment in the autophagy pathway. CYTO-ID® staining and Western blot results showed that GAX induced autophagy, upregulated the expression of Beclin-1, ATG5, and LC3-Ⅱ proteins, and downregulated the expression of p62 protein. This study suggests that GAX inhibits the proliferation, migration, and induces apoptosis of hepatocellular carcinoma cells by inducing autophagy, thereby significantly inhibiting tumor growth. GAX represents a promising adjuvant therapy for cancer treatment.

S100A12 is a member of S100 calcium-binding proteins with effect to promote inflammation in brain damage and stroke. However, the role of S100A12 in ischemia/reperfusion (I/R) remains to be clarified. This study aimed to explore the effect of S100A12 on I/R and discover the possible mechanism. Oxygen-glucose deprivation and reperfusion (OGD/R) was used to induce I/R injury model in vitro. Knockdown or overexpression of S100A12 was utilized to explore the role of S100A12 in I/R-induced inflammation and apoptosis. Results indicated that S100A12 expression was dramatically upregulated after OGD/R. Knockdown of S100A12 inhibited, while overexpression of S100A12 enhanced, the activation of ERK1/2 protein. OGD/R also triggered the occurrence of inflammation and oxidative stress, while these effects were blunted by S100A12 silencing and aggravated by S100A12 overexpression, and the presence of MAP kinase signaling system (ERK) inhibitor MK-8353 counteracted the effect of S100A12 overexpression. Besides, S100A12 silencing abolished, while its overexpression restored, the OGD/R-induced increased apoptosis rate and pro-apoptotic proteins expression. Similarly, ERK inhibitor MK-8353 reversed the effects of S100A12 overexpression. In conclusion, S100A12 promoted OGD/R-induced inflammation, oxidative stress and apoptosis via activation of ERK signaling in vitro., (© 2020 Japanese Society of Pathology and John Wiley & Sons Australia, Ltd.)

A series of disubstituted xanthones was obtained by cationic modification of xanthone's C2 and C7 with amine groups of different pKa values. Modified structures by using moieties with high pKa values had good antitumor activity according to the MTT assay, AO/EB staining and flow cytometry assay, especially bis-dimethylamine derivative (5a). Further study indicated that compound 5a had good binding activity to telomeric G-quadruplex DNA, as detected by using spectroscopy methods, melting profiles, polymerase chain reaction stop assay and molecular modeling study. The results suggested that the antitumor activity of 5a might be associated with its stabilization of G-quadruplex DNA, which could be developed as new G-quadruplex DNA stabilizer and potent antitumor agents., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Cutaneous squamous cell carcinoma (cSCC) accounts for 20% of non-melanoma skin cancer worldwide. MicroRNAs (miRNAs or miRs) are a subtype of non-coding RNA associated with the progression of various types of human cancer. MiR-186 has been demonstrated to act as an oncogene in human tumors. However, the role of miR-186 in cSCC remains unclear. The expression of miR-186 and apoptotic protease activating factor 1 (APAF1) was examined using reverse transcription-quantitative polymerase chain reaction, western blotting and immunofluorescence. The correlation between miR-186 and APAF1 was determined using a dual-luciferase assay. Mimics or inhibitors of miR-186 were transfected into A-431 cells to establish cell lines with overexpressed or knocked-down miR-186, respectively. EdU staining and colony formation assays were performed to detect cell proliferation. Transwell and wound-healing assays were performed to analyze cell invasion and migration, respectively. Hoechst staining and flow cytometry were performed to assess cell apoptosis and cell cycle distribution. MiR-186 expression was significantly increased, while APAF1 expression was significantly decreased in cSCC tissues compared with the controls. An miR-186 binding site was predicted in APAF1 and their expression was negatively correlated in cSCC tissues. Cell proliferation, invasion and migration were significantly enhanced in the miR-186-overexpressed A-431 cells and attenuated in miR-186 knockdown cells compared with the control. APAF1 expression was regulated by miR-186, while APAF1 knockdown significantly promoted cell invasion and inhibited cell apoptosis. In summary, the results of the present study indicate that miR-186 serves as an oncogene in cSCC by inhibiting APAF1. [ABSTRACT FROM AUTHOR]

• A novel series of xanthone derivatives were designed, synthesized and evaluated. • Compound 2a might be used as a potent quadruplex stabilizer and antitumor agent. • Effects of substitution on quadruplex binding and cytotoxicity have been discussed. A series of xanthone derivatives (2a - 2e) as telomeric quadruplex stabilizing ligands were designed, synthesized and evaluated by molecular docking, drug-likeness prediction, spectroscopy assay, melting profiles, PCR stop analysis. The ligands possessed the rigid aromatic xanthone scaffold and the flexible basic side chain with the appropriate length. Molecular docking and quadruplex binding property studies revealed that the ligands could stack on the external G-quartets, and the basic side chain could contribute to the stabilizing ability. Introduction of a tertiary amine group with high p K a value and small steric hinderance was favorable for improving the binding ability. Such as bis-dimethylamine derivative (2a), exhibited stronger binding ability and higher affinity toward G-quadruplex than duplex DNA. Cytotoxicity assay showed that the xanthone derivatives could inhibit the viabilities of HeLa, MCF-7, SGC-7901 and A549 cells in different degrees. AO/EB staining and flow cytometry assay indicated that the cytotoxicity of xanthones was related to induced apoptosis. These results suggested that the ligands could be further studied as potential new G-quadruplex stabilizers and potent antitumor agents, especially 2a. [Display omitted] [ABSTRACT FROM AUTHOR]

The study is aimed at studying the effects of soluble and insoluble oat β-glucan on colon carcinogenesis in mice. One hundred and twenty male Kunming mice were divided into normal control (NC), model control (MC), high doses (100 mg/kg body weight) of soluble (H-SOG) and insoluble β-glucan (H-IOG), and low doses (50 mg/kg body weight) of soluble (L-SOG) and insoluble β-glucan (L-IOG) groups. The mice except those in the NC group were given subcutaneous injections of DMH to induce colon cancer. The bile acid content was significantly reduced but the colonic short-chain fatty acid content was enhanced (p < .05) in mice administered β-glucan, moreover, the tumor cells apoptosis was significantly promoted (p < .05), compared to the MC group. The effect of high doses β-glucan was better than the low doses, but there is little difference between the SOG and IOG. Results suggested that both SOG and IOG exert favorable effects in preventing colon cancer in a doses-dependent manner while the specific mechanism maybe different. [ABSTRACT FROM AUTHOR]

Background: In a swine model of acute myocardial infarction (AMI), Statins can enhance the therapeutic efficacy of mesenchymal stem cell (MSCs) transplantation. However, the mechanisms remain unclear. This study aims at assessing whether atorvastatin (Ator) facilitates the effects of MSCs through activation of nitric oxide synthase (NOS), especially endothelial nitric oxide synthase (eNOS), which is known to protect against ischemic injury. Methods and Results: 42 miniswines were randomized into six groups (n = 7/group): Sham operation; AMI control; Ator only; MSC only, Ator+MSCs and Ator+MSCs+NG-nitrol-L-arginine (L-NNA), an inhibitor of NOS. In an open-heart surgery, swine coronary artery ligation and reperfusion model were established, and autologous bone-marrow MSCs were injected intramyocardium. Four weeks after transplantation, compared with the control group, Ator+MSCs animals exhibited decreased defect areas of both “perfusion” defined by Single-Photon Emission Computed Tomography (−6.2±1.8% vs. 2.0±5.1%, P = 0.0001) and “metabolism” defined by Positron Emission Tomography (−3.00±1.41% vs. 4.20±4.09%, P = 0.0004); Ejection fraction by Magnetic Resonance Imaging increased substantially (14.22±12.8% vs. 1.64±2.64%, P = 0.019). In addition, indices of inflammation, fibrosis, and apoptosis were reduced and survivals of MSCs or MSC-derived cells were increased in Ator+MSCs animals. In Ator or MSCs alone group, perfusion, metabolism, inflammation, fibrosis or apoptosis were reduced but there were no benefits in terms of heart function and cell survival. Furthermore, the above benefits of Ator+MSCs treatment could be partially blocked by L-NNA. Conclusions: Atorvastatin facilitates survival of implanted MSCs, improves function and morphology of infarcted hearts, mediated by activation of eNOS and alleviated by NOS inhibitor. The data reveal the cellular and molecular mechanism for anti-AMI therapy with a combination of statin and stem cells. [ABSTRACT FROM AUTHOR]

Objective To evaluate the influence of overexpression HGF on the healing of traumatic ulcer of oral mucosa of mice. Material and methods Mice were divided into two groups: wild type C57BL6(WT) and HGF high expression transgenic (HGF-Tg) mice. Traumatic ulcer of all mice were made by number 15 scalpel blade. Mice were sacrificed after 5 days and the inflammation score and expression of TNFα, IFNγ, c-Met, apoptosis (TUNEL) and 40 serum inflammation cytokines were estimated. Results HGF-Tg mice presented a lower inflammation score (p = 0.011), Serum TNFα expression in HGF-Tg ulcers is 1.3 times than WT ulcer and the difference is statistical significance ( t test, p = 0.003). Serum c-Met protein in HGF-Tg mice were significantly higher than WT mice ( t test, p = 0.004). No statistical difference was observed in the serum IFNγ between WT ulcer and HGF-Tg ulcer ( t test, p = 0.268). TNFα positive cytoplasm expression cells in connective tissue of HGF-Tg mice is significantly lower than that of WT group ( t test, p = 0.029). C-Met positive cytoplasm expression cells in both epithelium and connective tissue of HGF-Tg group is significantly higher than that of WT group ( t test, p = 0.040, p = 0.000). Samples in HGF-Tg group showed a lower number of positive cells of epithelium TUNEL staining compared with that in the WT group ( t test, p = 0.035). Conclusions HGF exhibited anti-inflammatory potential in oral traumatic ulcer through the reduction of epithelial apoptosis, connective tissue TNFα expression and induction of c-Met expression. [ABSTRACT FROM AUTHOR]