Project description:This experiment includes treatment of human pulmonary fibroblasts obtained from IPF patients with metformin. Since, we would like to investigate the transcriptome profile of these samples following metformin treatment. There will be two groups consist of four samples each. First group treated with metformin for 72 hours, while the second group treated with vehicle.
Project description:To examine the effect of metformin on lung cancer biology, human lung H226 and H1299 squamous cell carcinoma cell-lines were grown in RPMI-1640 medium with 10% v/v fetal bovine serum and with or without 15 uM metformin hydrochloride for 7-8 days. Medium (with any metformin) was replaced every two days. Paired cultures with and without metformin were grown and maintained in parallel. Three separate paired cultures, all seeded with same stock of frozen cells, were grown.
Project description:The DNA regions whose chromatin accessibility was significantly altered by metformin were investigated in lung cancer cells by ATAC-seq. Metformin decreased chromatin accessibility in 159 regions but increased it in 124 sites. Most regions were outside of gene promoters defined as TSS ± 3kb. The most suppressed region was 4,667bp downstream of CHAC1, and the most enhanced site was 15,111bp downstream of ERRFI1.
Project description:To examine the effect of metformin on lung cancer biology, human lung A549 adenocarcinoma, H460 large cell carcinoma, and H226 and H1299 squamous cell carcinoma cell-lines were grown in RPMI-1640 medium with 10% v/v fetal bovine serum and with or without 15 uM metformin hydrochloride for 7-8 days. Medium (with any metformin) was replaced every two days. Paired cultures with and without metformin were grown and maintained in parallel. Three separate paired cultures, all seeded with same stock of frozen cells, were grown. Cells, within 15%-95% confluence range, were harvested by scraping. Total RNA from cells was prepared using Norgen Biotek® Total RNA Isolation kit (with on-column DNAse I treatment). All RNA integrity number (RIN) values were greater than 8.2.
Project description:Analysis of Metfromin induced changes in the lung cells of Mycobacterium tuberculosis infected mouse at gene expression level. The hypothesis tested in the present study was whether metformin has any effect on the host immune response in Mycobacterium tuberculosis infected mice? Results provide important information on the effect of metformin on the inflammatory response and immune activation associated with mycobacterial infection. In conclusion, Metfromin normalizes the chronic inflammation associated with Mycobacterium tuberculosis infection.
Project description:The DNA regions that were associated with the metformin altered expressions of H3K4me3 and/or H3K27me3 were investigated by ChIP-seq, and the regions significantly up- or down-regulated by metformin were identified in H1299 cells. Over 80% of H3K4me3 were discovered within promoter regions, but less than 10% of H3K27me3 were observed in this area. Metformin significantly downregulated H3K4me3 level at the promoter regions of 1113 genes, but upregulated levels in the promoter regions of 1492 genes. Gene enrichment analysis revealed that abundant cell cycle related genes such as PIK3C3, CCNB2, CDK1, CDK6, and E2F8 were upregulated, while certain cell cycle suppressor genes such as CDKN1A (p21) and DDIT4 were downregulated. No specific biological functional cluster was detected in the H3K27me3 up- or downregulated genes.
Project description:The genes whose transcription was significantly altered by metformin were investigated in H1299 cells by RNA-seq. Significantly up- or down-regulated genes were identified by cutoff value of |FC| >= 1.5. Metformin significantly downregulated mRNA levels of 499 genes while upregulating 615. KEGG (Kyoto Encyclopedia of Genes and Genomes) gene enrichment analysis revealed that the highly ranked clusters were cell cycle, apoptosis, cellular senescence and p53 signaling pathways. Multiple cell cycle checkpoint genes such as CCNA, CCNB, CCNE, CDK1, E2F2, E2F6, and E2F8 were downregulated, while certain cell cycle inhibitors such as CDKN2B (p15), CDKN1A (p21), DDIT4, and GADD45 were upregulated.