Inhibitory effect of gemcitabine and brucine on MDA MB-231human breast cancer cells

Combination of natural compounds and cytotoxic drugs represents a logical therapeutic approach for breast cancer. Brucine, a natural plant alkaloid is reported to possess cytotoxic, anti-inflammatory and anti-cancer activities. Gemcitabine is a nucleoside analog, commonly used in the treatment of several solid tumors, including breast cancer. In the present study we have examined the anticancer effect of brucine and gemcitabine on MDA MB-231 human breast cancer cells. Cell proliferation was assessed using MTT assay. Soft agar assay was used to evaluate the in-vitro clonogencity of MDA MB-231 cells. Cell migration was determined by in-vitro scratch assay and expression of p65 (NF-kB subunit) was evaluated by western blot analysis. Combination treatment with brucine and gemcitabine resulted in a significant inhibition of cell proliferation than either brucine or gemcitabine alone. Cells treated with combination of brucine and gemcitabine showed additive inhibition in colony formation and cell migration than treated with individual agents. The cells treated with brucine at 300 µM showed a significant decrease in p65-NF-kB expression but in combination treatment there was no additive inhibition of p65 expression compared to brucine treated cells. Overall, our results suggested that brucine in combination with gemcitabine showed supra-additive anticancer effects in MDA MB-231 cells and warrants further in-vivo studies in experimental animal models.

Dietary Lutein Attenuates Bleomycin-Induced Pulmonary Fibrosis in Mice

Idiopathic Pulmonary Fibrosis is a chronic lung disease characterized by lung inflammation and progression to fibrosis. Anti-fibrotic efficacy of lutein was studied in mouse model of bleomycin induced pulmonary fibrosis. The mice were orally administrated with 100 or 200 mg/kg of lutein three days prior to intra-tracheal instillation of bleomycin on day 0 and continued up to day 21. Total cell counts and protein levels in branchoalveolar lavage fluid and superoxide dismutase, and hydroxyproline levels were evaluated. Treatment with lutein at 100 and 200 mg/kg prevented bleomycin induced mortality and body weight loss. Lutein administration attenuated bleomycin induced increase of total and differential cell count and myeloperoxidase, inflammatory cell infiltration in lung tissue. Similarly lutein administration restored superoxide dismutase activity lowered by bleomycin instillation. The combined results revealed the protective effect of lutein by suppressing the inflammation, oxidative stress and enhancing the antioxidant potential which may helps in pulmonary fibrosis

ORAL ADMINISTRATION OF BOSENTAN ATTENUATES THE BLEOMYCIN INDUCED PULMONARY FIBROSIS IN MICE

Idiopathic Pulmonary Fibrosis (IPF) is characterized by alveolitis with epithelial, endothelial damage leading to fibrosis. As signaling of endothelin-1 was involved in IPF, the effect of bosentan a non-specific endothelin receptor antagonist was determined in a mouse model of bleomycin induced pulmonary fibrosis. In the present study, mice were instilled with intra-tracheal instillation of bleomycin (0.05U) and were administered with bosentan at 100 mg/kg body weight. The treatment with bosentan significantly (p ≤ 0.05) prevented bleomycin induced mortality and loss of body weight. On day 7, bosentan significantly (p ≤ 0.05) attenuated bleomycin induced increase of total and differential inflammatory cell counts, total proteins, edema, MPO activity and inflammatory cell infiltration in lung tissue. The activities of superoxide dismutase and catalase were restored by bosentan treatment which lowered in bleomycin administered mice. Bosentan treatment significantly attenuated bleomycin induced increase in fibrosis score, collagen deposition and hydroxyproline levels. On day 21, treatment with bosentan significantly attenuated α-smooth muscle actin and collagen-I gene expression levels and matrix metalloproteinases 2 and 9 activities. The results revealed that bosentan exerted enhanced protection against bleomycin induced inflammation and fibrosis

Spatially restricted drivers and transitional cell populations cooperate with the microenvironment in untreated and chemo-resistant pancreatic cancer

Pancreatic ductal adenocarcinoma is a lethal disease with limited treatment options and poor survival. We studied 83 spatial samples from 31 patients (11 treatment-naïve and 20 treated) using single-cell/nucleus RNA sequencing, bulk-proteogenomics, spatial transcriptomics and cellular imaging. Subpopulations of tumor cells exhibited signatures of proliferation, KRAS signaling, cell stress and epithelial-to-mesenchymal transition. Mapping mutations and copy number events distinguished tumor populations from normal and transitional cells, including acinar-to-ductal metaplasia and pancreatic intraepithelial neoplasia. Pathology-assisted deconvolution of spatial transcriptomic data identified tumor and transitional subpopulations with distinct histological features. We showed coordinated expression of TIGIT in exhausted and regulatory T cells and Nectin in tumor cells. Chemo-resistant samples contain a threefold enrichment of inflammatory cancer-associated fibroblasts that upregulate metallothioneins. Our study reveals a deeper understanding of the intricate substructure of pancreatic ductal adenocarcinoma tumors that could help improve therapy for patients with this disease

Profiling Single Cancer Cells with Volatolomics Approach

Summary: Single-cell analysis is a rapidly evolving to characterize molecular information at the individual cell level. Here, we present a new approach with the potential to overcome several key challenges facing the currently available techniques. The approach is based on the identification of volatile organic compounds (VOCs), viz. organic compounds having relatively high vapor pressure, emitted to the cell's headspace. This concept is demonstrated using lung cancer cells with various p53 genetic status and normal lung cells. The VOCs were analyzed by gas chromatography combined with mass spectrometry. Among hundreds of detected compounds, 18 VOCs showed significant changes in their concentration levels in tumor cells versus control. The composition of these VOCs was found to depend, also, on the sub-molecular structure of the p53 genetic status. Analyzing the VOCs offers a complementary way of querying the molecular mechanisms of cancer as well as of developing new generation(s) of biomedical approaches for personalized screening and diagnosis. : Analytical Chemistry; Sensor; Biological Sciences; Cancer Systems Biology Subject Areas: Analytical Chemistry, Sensor, Biological Sciences, Cancer Systems Biolog

Epigenetic and transcriptomic characterization reveals progression markers and essential pathways in clear cell renal cell carcinoma

Tumour heterogeneity in clear cell renal cell carcinoma (ccRCC) remains to be investigated. Here, the integration of spatial omics, transcriptional and chromatin accessibility profiling at the single-nucleus level and bulk proteogenomics data reveal markers and pathways important for ccRCC