Project description:We investigated the effects of the crude extract of a South African medicinal plant, Cotyledon orbiculata, on cell survival of colon (HCT116) cancer cell lines. Using RNASeq, we discovered that the extract interfered with mRNA regulatory pathways. Here, we found that the extract of Cotyledon orbiculata, a South African medicinal plant, had an anti-proliferative effect in cancer cells, mediated by apoptosis induced by alternative splicing of hnRNPA2B1 and BCL2L1.
Project description:We investigate the molecular targets and pathways that the neem extracts and the associated compounds act through, to bring about tumor suppression by using a genome-wide functional pooled shRNA screen on head and neck squamous cell carcinoma cell line treated with crude neem leaf extracts. We analyzed differences in global clonal sizes of the shRNA-infected cells cultured under no treatment and treatment with neem leaf extract conditions, assayed using next-generation sequencing. We further analyzed differences in gene expression in HSC-4 cells, upon treatment with neem leaf extract in a time-dependent manner, followed by a time-dependent rescue. Our results indicate that neem extract simultaneously affects various important molecular signaling pathways in head and neck cancer cells, some (TGF-β/HSF-1) of which may be therapeutic targets for this devastating tumor.
Project description:Biological replicates: 393-395 (crude extract): protein extraction 399-401 (40% AS): protein extraction+40% AS precipitation 405-407 (crude extract+MOAC): protein extraction+phosphoprotein enrichment by MOAC 408-410 (40% AS+MOAC): protein extraction+40% AS precipitation+phosphoprotein enrichment by MOAC -determination of protein concentration by 2D-Quant Kit -LC-MS with LTQ Orbitrap Velos: nanoLC, precursor scan Orbitrap, fragmentation CID, fragment scan LIT -Thermo Proteome Discoverer 1.3 with phosphoRS 1.0 -Mascot 2.3 -database TAIR 10 -Scaffold 4 Technical replicates two measurements each (e.g. 393, 393_2)
Project description:To assess the influences of a crude medicinal herb extract (MHE) on the immune composition and function in inguinal white adipose tissue (iWAT), we isolated iWAT stromal vascular fractions (SVFs) from control (PBS) and MHE-treated mice and performed RNA-seq analysis. iWAT SVF of mice treated with MHE was defined as the treatment group. Phosphate-buffered saline (PBS) treatment was used as the control group. Then RNA-Seq experiment was performed by OE Biotech Co., Ltd. (Shanghai, China) to analyze gene expression changes in iWAT SVF.
Project description:Worldwide, more than 1 billion people are affected by infestations with soil-transmitted helminths and also in veterinary medicine helminthiases are a severe thread to livestock due to emerging resistances against the common anthelmintics. Proanthocyanidins have been increasingly investigated for their anthelmintic properties, however, except for an interaction with certain proteins of the nematodes, not much is known about their mode of action. To investigate the anthelmintic activity on a molecular level, a transcriptome analysis was performed in Caenorhabditis elegans after treatment with purified and fully characterized oligomeric procyanidins (OPC). The OPCs had previously been obtained from a hydro-ethanolic (1:1) extract from the leaves of Combretum mucronatum, a plant which is traditionally used in West Africa for the treatment of helminthiasis, therefore, also the crude extract was included in the study. Significant changes in differential gene expression were observed mainly for proteins related to the intestine, many of which were located extracellularly or within cellular membranes. Among the up-regulated genes, several hitherto undescribed orthologues of structural proteins in humans were identified, but also genes that are potentially involved in the worms’ defense against tannins. For example, T22D1.2, an orthologue of human basic salivary proline-rich protein (PRB) 2, and numr-1 (nuclear localized metal responsive) were found to be strongly up-regulated. Down-regulated genes were mainly associated with lysosomal activity, glycoside hydrolysis or the worms’ innate immune response. No major differences were found between the groups treated with purified OPCs versus the crude extract. Investigations using GFP reporter gene constructs of T22D1.2 and numr-1 corroborated the intestine as the predominant site of the anthelmintic activity.
Project description:Using LC-MS/MS, we analyzed ECM-enriched samples obtained from 1) human triple-negative breast cancer samples and matched adjacent normal mammary gland tissues, and 2) disease-free omentum from patient with non-metastatic ovarian cancer and high-grade- serous-ovarian- cancer-derived omental metastasis samples. We conducted the LC-MS/MS analysis on peptides obtained after solubilizing ECM-enriched samples using different methods (crude ECM extract, urea-soluble extract, urea-insoluble extract) and submitted to basic-reversed- phase separation or not.
Project description:Triton X-100 soluble crude extract from BCECs displaying limited blood-brain barrier functions was fractionated into 5 fractions (F0, F25, F50, F75) with increasing concentrations of acetonitrile (0%, 25%, 50%, 75%). This is the merging of triplicate dataset of subfraction # F0.