The Role of Autophagy in Tumor Immune Infiltration in Colorectal Cancer.

Objective: This study is aimed at exploring the association between autophagy and tumor immune infiltration (TII) in colorectal cancer (CRC). Methods and Materials: We downloaded the transcriptome profiling and clinical data for CRC from The Cancer Genome Atlas (TCGA) database and obtained the normal colon transcriptome profiling data from Genotype-Tissue Expression Project (GTEx) database. The list of autophagy-related signatures was obtained from the Human Autophagy Database. We isolated the autophagy-related genes from the CRC gene expression matrix and constructed an autophagy-related prognostic (ARP) risk model. Then, we constructed a multiROC curve to validate the prognostic ability of the ARP risk model. CIBERSORT was used to determine the fractions of 22 immune cells in each CRC sample, and the association between these TII cells and CRC clinical variables was further investigated. Finally, we estimated the association of 3 hub-ARP signatures and 20 different types of TII cell distribution. Results: We classified 447 CRC patients into 224 low-risk and 223 high-risk patients using the median ARP risk score. According to the univariate survival test results, except for gender (P = 0.672), age (P = 0.008), cancer stage, and pathological stage T, M, and N were closely correlated with the prognosis of CRC patients (P < 0.001). Multivariate survival analysis results indicate that age and rescore were the only independent prognostic indicators with significant differences (P < 0.05). After merging the immune cell distribution (by CIBERSORT) with the CRC clinical data, the results indicate that activated macrophage M0 cells exhibited the highest clinical response, which included cancer stage and stage T, N, and M. Additionally, six immune cells were closely associated with cancer stage, including regulatory T cells (Tregs), gamma delta T cells, follicular helper T cells, activated memory CD4 T cells, activated NK cells, and resting dendritic cells. Finally, we evaluated the correlation of ARP signatures with TII cell distribution. Compared with the other correlation, NRG1 and plasma cells (↑), risk score and macrophage M1 (↑), NRG1 and dendritic cell activated (↑), CDKN2A and T cell CD4 memory resting (↓), risk score and T cell CD8 (↑), risk score and T cell CD4 memory resting (↓), and DAPK1 and T cell CD4 memory activated (↓) exhibited a stronger association (P < 0.0001). Conclusions: In summary, we explored the correlation between the risk of autophagy and the TII microenvironment in CRC patients. Furthermore, we integrated different CAR signatures with tumor-infiltrating immune cells and found robust associations between different levels of CAR signature expression and immune cell infiltrating density.

Isorhamnetin protects endothelial cells model CRL1730 from oxidative injury by hydrogen peroxide.

To explore the protective effect and mechanism of isorhamnetin against oxidative injury caused by H2O2 to endothelial cell strain CRL1730 of human umbilical vein. H2O2 and endothelial cell strain CRL1730 were used, as a model of injured endothelial cell. Three levels of crude drugs areorhamnetin, 22.8µg/ml, 11.4µg/mL and 5.7µg/mL was added to the injured cell strain CRL1730 respectively. The cell injury was measured in terms of necrotic rate, quantities of von Wilebr and factor (vWf) and thrombomodulin (TM), lactate dehydrogenase (LDH) and intracellular free calcium ions through flow cytometry, ELISA, fluorescent spectrometer and laser scanning confocal microscopy respectively. Isorhamnetins @ 11.4µg/mL and 5.7µg/mL has significantly decreased EC necrotic rate, while the increased vWf concentration due to oxidant (200mol/L of H2O2) was significantly decreased by 5.7µg/mL versus 11.4 and 22.8µg/mL isorhamnetin. Also, the increased in TM and LDH in injured cells was reversed to normal level with 5.7 to 11.4µg/mL isorhamnetin. These results suggest that isorhamnetin protect the integrity of cell membranes. Similarly, H2O2 treatment of cells elicited the release of intracellular calcium, however, 5.7µg/mL and 11.4µ g/mL isorhamnetin dramatically inhibited transient release of intracellular calcium. This suggests that isorhamnetin, at lower concentration, could inhibit the IP3-sensitive calcium pool from releasing calcium, protecting VECs from injury by H2O2. Traditional Chinese herbs, hippophaerhamnoides have been recognized as safe and as a source of flavonoids, with strong cardiovascular protection. The results of this study revealed that isorhamnetin produce a strong effect on some targetspresent in ECs and thus, provide a basis for the future work targeted towards endothelial cells protection.

Protection and mechanism of total flavone of Hippophae rhamnoides on vascular endothelial cells / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the protection of total flavones of Hippophae Rhamnoides (TFH) on vascular endothelial cells (VECs).</p><p><b>METHODS</b>Human umbilical VECs (ECV304) were used. The vascular endothelial injured cell model was prepared using hydrogen dioxide (H2O2). The cell apoptosis rate and changes of mean fluorescence intensity were detected using flow cytometry (FCM). The Caspase-3 activity in VECs was detected by Western blot.</p><p><b>RESULTS</b>VEC apoptosis was induced by 200 micromol/L H2O2. TFH in different concentrations (400, 200, and 100 microg/mL) could significantly lower the cell apoptosis rate induced by H2O2 respectively (all P < 0.05), and obviously inhibit Caspase-3 activities (all P < 0.01).</p><p><b>CONCLUSIONS</b>TFH could fight against H2O2 injured VECs apoptosis. Lowering the Caspase-3 expression was one of its mechanisms in protecting VECs.</p>