Metabolites contributing to the difference between healthy control and high IP₃R patient group.

<p>Metabolites contributing to the difference between healthy control and high IP₃R patient group.</p

An overview of the workflow performed for the serum metabolic profiling of breast cancer patients using ¹H NMR spectroscopy.

<p>An overview of the workflow performed for the serum metabolic profiling of breast cancer patients using ¹H NMR spectroscopy.</p

Inhibition of IP₃R in MCF -7 breast cancer cells effects metabolism.

<p>Glucose uptake in MCF-7 cells was analyzed using NBDG a fluorescently labeled deoxy glucose analogue as a probe for detection of glucose taken up by cultured cells. Quantitative estimation of glucose uptake, using a cell based assay kit, was performed as per instructions provided by the manufacturer (Cayman, USA). Cells were plated in 96-well plates and treated with 25 μM XeC for 24 hours or with siC (non-targeted siRNA) or siIP₃R2 or siIP₃R3 (72 hours,Fig7A and 7D) Representative graph in MCF-7 cells. (Fig 7B and 7E) Representative graph showing percentage of glucose uptake as estimated using cell based assay in MDA MB-231 cells. (Fig7C and 7F) Representative graph showing percentage of glucose uptake as estimated using cell based assay in MCF 10A cells. RNA was extracted from treated and untreated cells and cDNA was prepared (Fig 7G) RT profiler PCR array for glucose as well as mitochondrial metabolism genes was performed using cDNA prepared from mRNA of MCF-7 cells. Data represent mean ±SEM. *p< 0.05, ***p< 0.001 compared to vehicle.</p

Representative box-cum-whisker plots for significant metabolites of healthy control vs high IP₃R patient group (with p>0.05).

<p>Relative integral areas are calculated from CPMG spectra and plotted as a.u.</p

Score Plot Generated From PCA And PLS-DA Analysis Of NMR Spectra Of Serum From Healthy Control And Patient Groups.

<p>(A) PCA and (D) PLS-DA score plot generated for healthy control vs. patient group, (B) PCA and (E) PLS-DA score plot generated for healthy control and high IP₃R group, (C) PCA and (F) PLS-DA score plot generated for healthy control and low IP₃R group.</p

Analysis of expression of Inositol 1, 4, 5-trisphosphate receptors type 2 (IP₃R2) and type 3 (IP₃R3) in tumor tissue of breast cancer patients.

<p>Relative mRNA expression of (A) IP₃R1, (B) IP₃R2 (p<0.001) and (C) IP₃R3 (p<0.001) from tumoral and extra-tumoral tissues of breast cancer patients was analyzed using Real -time PCR. Analysis of tumoral and extra-tumoral tissues was done by incubating the tissue sections with antibodies against (D) IP₃R2 and (E) IP₃R3, and detected using DAB staining.</p

An integrated analysis based on Metaboanalyst software (pathway tool) for a simplified view of contributing pathways.

<p>The panel shows a view of metabolism in cancers depicting glycolysis and Krebs cycle which are modified to different processes like lipid and amino acid synthesis to meet the requirement of proliferating cells. (Metabolites depicted with green/red are decreased/increased in the present study).</p

¹H NMR Metabolomics Reveals Association of High Expression of Inositol 1, 4, 5 Trisphosphate Receptor and Metabolites in Breast Cancer Patients - Fig 5

<p>Score plot and corresponding loading plot generated from PLS-DA analysis between (A) healthy control and patient group, (B) healthy control and high IP₃R group, (C) healthy control and low IP₃R group.</p