Antitumor cell activity in glioblastoma regulated by inhibiting triggering receptor expressed on myeloidcells 2 ScienceAdvances CellCellPress
-catenin , anti-phosphorylated ERK and anti-ERK. Antibodies against Wnt, p53 and p21 were obtained from Santa Cruz Biotechnology . Anti-TREM2 and TREM2-Ig were obtained from R&D Systems . Anti-EpCAM and anti--tubulin antibodies were purchased from Abcam and Sigma, respectively. Horseradish peroxidase -conjugated anti-goat IgG and anti-rabbit IgG were used as secondary antibodies for Western blot analysis.Whole colon tissue was removed and dissected away from any attached connective tissues.
1 vector alone or the same vector containing TREM2 cDNA using Lipofectamine 2000 as described in the manufacturer’s protocol. At 4 h post-transfection, the DNA-containing medium was replaced with fresh medium supplemented with 10% FBS, and the cells were incubated at 37 °C in a humidified 5% CO4.7. Reverse Transcription-polymerase Chain Reaction reagent according to the manufacturer’s instructions.
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Exploring the role of pyroptosis in shaping the tumor microenvironment of colorectal cancer by bulk and single-cell RNA sequencing - Cancer Cell InternationalBackground Emerging studies have shown that pyroptosis plays a non-negligible role in the development and treatment of tumors. However, the mechanism of pyroptosis in colorectal cancer (CRC) remains still unclear. Therefore, this study investigated the role of pyroptosis in CRC. Methods A pyroptosis-related risk model was developed using univariate Cox regression and LASSO Cox regression analyses. Based on this model, pyroptosis-related risk scores (PRS) of CRC samples with OS time | 0 from Gene Expression Omnibus (GEO) database and The Cancer Genome Atlas (TCGA) database were calculated. The abundance of immune cells in CRC tumor microenvironment (TME) was predicted by single-sample gene-set enrichment analysis (ssGSEA). Then, the responses to chemotherapy and immunotherapy were predicted by pRRophetic algorithm, the tumor immune dysfunction and exclusion (TIDE) and SubMap algorithms, respectively. Moreover, the Cancer Therapeutics Response Portal (CTRP) and PRISM Repurposing dataset (PRISM) were used to explore novel drug treatment strategies of CRC. Finally, we investigated pyroptosis-related genes in the level of single-cell and validated the expression levels of these genes between normal and CRC cell lines by RT-qPCR. Results Survival analysis showed that CRC samples with low PRS had better overall survival (OS) and progression-free survival (PFS). CRC samples with low PRS had higher immune-related gene expression and immune cell infiltration than those with high PRS. Besides, CRC samples with low PRS were more likely to benefit from 5-fluorouracil based chemotherapy and anti-PD-1 immunotherapy. In novel drug prediction, some compounds such as C6-ceramide and noretynodrel, were inferred as potential drugs for CRC with different PRS. Single-cell analysis revealed pyroptosis-related genes were highly expressed in tumor cells. RT-qPCR also demonstrated different expression levels of these genes between normal and CRC cell lines. Conclusions Taken together, this s
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