Lipid-rich Cells Research Articles

Lipid Production by Rhodotorula glutinis in Continuous Cultivation with a Gravity Sedimentation System.

Lipid accumulation is generally believed to be a partially growth-coupled biochemical process that results in differences in lipid content between different cells. To separate lipid-rich cells and increase the cellular biomass in bioreactors during the cultivation of the oleaginous yeasts, a gravity sedimentation system (GSS) is coupled to a bioreactor. The dilution rate (D) and the ratio of the outflow rate from the outlet of the GSS to the inflow rate into the bioreactor (B) were evaluated. The biomass in the bioreactor with GSS increased by 16.3% and 30.6% at D values of 0.05h-1 (B = 0.25) and 0.02h-1 (B = 0.5), respectively. Interestingly, cells containing 39.3% lipids were obtained from the outlet of the GSS (D = 0.02h-1, B = 0.5), and the lipid content increased by 7.8% compared to that of the bioreactor. The results indicated that use of a GSS is a very effective method for increasing the cell concentration and separation of lipid-rich cells.

PSMD1 and PSMD2 regulate HepG2 cell proliferation and apoptosis via modulating cellular lipid droplet metabolism

BackgroundObesity and nonalcoholic steatohepatitis (NASH) are well-known risk factors of hepatocellular carcinoma (HCC). The lipid-rich environment enhances the proliferation and metastasis abilities of tumor cells. Previous studies showed the effect of the ubiquitin–proteasome system (UPS) on tumor cell proliferation. However, the underlying mechanism of UPS in regulating the proliferation of lipid-rich tumor cells is not totally clear.ResultsHere, we identify two proteasome 26S subunits, non-ATPase 1 and 2 (PSMD1 and PSMD2), which regulate HepG2 cells proliferation via modulating cellular lipid metabolism. Briefly, the knockdown of PSMD1 and/or PSMD2 decreases the formation of cellular lipid droplets, the provider of the energy and membrane components for tumor cell proliferation. Mechanically, PSMD1 and PSMD2 regulate the expression of genes related to de novo lipid synthesis via p38-JNK and AKT signaling. Moreover, the high expression of PSMD1 and PSMD2 is significantly correlated with poor prognosis of HCC.ConclusionWe demonstrate that PSMD1 and PSMD2 promote the proliferation of HepG2 cells via facilitating cellular lipid droplet accumulation. This study provides a potential therapeutic strategy for the treatment of lipid-rich tumors.

A rapid sampling technique for isolating highly productive lipid-rich algae strains from environmental samples

Strain selection and isolation of lipid-rich microalgae are among the two most important steps for screening isolates with maximum biofuel productivity. In this work, we introduce a novel direct sampling technique that allows native strains to be selected for rapid growth under defined conditions followed by direct selection of product-rich species, two desirable characteristics of algae for mass culture. This sampling strategy directly selects the lipid-rich strains visualized under an inverted fluorescence microscope using an X-Y-Z micromanipulator. The enrichment step can be manipulated to select for strains with specific technological applications. Direct sampling of lipid-rich cells avoids the tedious task of screening isolates while using relatively inexpensive equipment.

Tumor Cell Subtypes Based on the Intracellular Hormonal Activity in KCNJ5-Mutated Aldosterone-Producing Adenoma.

Aldosterone-producing adenomas (APAs) harbor marked intratumoral heterogeneity in terms of morphology, steroidogenesis, and genetics. However, an association of biological significance of morphologically identified tumor cell subtypes and genotypes is virtually unknown. KCNJ5 mutation is most frequently detected and generally considered a curable phenotype by adrenalectomy. Therefore, to explore the biological significance of KCNJ5 mutation in APA based on intracellular hormonal activities, 35 consecutively selected APAs (n=18; KCNJ5 mutated, n=17; wild type) were quantitatively examined in the whole tumor areas by newly developed digital image analysis incorporating their histological and ultrastructural features (14 cells from 2 KCNJ5-mutated APAs and 15 cells from 1 wild type) and CYP11B2 immunoreactivity. Results demonstrated that KCNJ5-mutated APAs had significantly lower nuclear/cytoplasm ratio and more abundant clear cells than wild type. CYP11B2 immunoreactivity was not significantly different between these genotypes, but a significant correlation was detected between the proportion of clear cells and CYP11B2 immunoreactivity in all of the APAs examined. CYP11B2 was predominantly immunolocalized in clear cells in KCNJ5-mutated APAs. Quantitative ultrastructural analysis revealed that KCNJ5-mutated APAs had significantly more abundant and smaller-sized mitochondria with well-developed cristae than wild type, whereas wild type had more abundant lipid droplets per unit area despite the small number of the cases examined. Our results did provide the novel insights into the morphological features of APA based on their biological significance. KCNJ5-mutated APAs were characterized by predominance of enlarged lipid-rich clear cells possibly resulting in increased neoplastic aldosterone biosynthesis.

Effects of indole-3-carbinol on steroid hormone profile and tumor progression in a mice model of canine inflammatory mammarycancer

BackgroundIndole-3-carbinol, derived from Cruciferous vegetables is an estrogen receptor antagonist considered a preventive agent that is naturally present in diet. There are no previous studies on its effects in human inflammatory breast cancer or canine inflammatory mammary cancer that is the most aggressive type of breast cancer.MethodsThe aim of this study was to analyze the effect of indole-3-carbinol on a SCID mice xenograft model of canine inflammatory mammary cancer, using equivalent human oral dose as a preventive therapy in humans for 3 weeks.ResultsIndole-3-carbinol treatment decreased tumor proliferation and increased apoptosis, although tumor embolization and liver metastasis were observed in some animals. There was a characteristic subpopulation of lipid-rich cells and increased contents of select steroid hormones in tumor homogenates and serum.ConclusionsOur data reveal for the first time that the ingestion of indole-3-carbinol, as administered, diminishes proliferation and increases apoptosis of tumor cells in an experimental model of inflammatory breast cancer, although this effect could not be enough to avoid the appearance of tumor embolization and metastasis. Future clinical trials will be needed to clarify the usefulness of indole-3-carbinol in this cancer and to understand the molecular mechanisms involved.

Detecting Phenotypically Resistant Mycobacterium tuberculosis Using Wavelength Modulated Raman Spectroscopy.

Raman spectroscopy is a non-destructive and label-free technique. Wavelength modulated Raman (WMR) spectroscopy was applied to investigate Mycobacterium tuberculosis cell state, lipid rich (LR) and lipid poor (LP). Compared to LP cells, LR cells can be up to 40 times more resistant to key antibiotic regimens. Using this methodology single lipid rich (LR) from lipid poor (LP) bacteria can be differentiated with both high sensitivity and specificity. It can also be used to investigate experimentally infected frozen tissue sections where both cell types can be differentiated. This methodology could be utilized to study the phenotype of mycobacterial cells in other tissues.

Enhanced Methodologies for Detecting Phenotypic Resistance in Mycobacteria.

Lipid droplets found in algaeand other microscopic organisms have become of interest to many researchers partially because they carry the capacity to produce bio-oil for the mass market. They are of importance in biology and clinical practice because their presence can be a phenotypic marker of an altered metabolism, including reversible resistance to antibiotics, prompting intense research.A useful stain for detecting lipid bodies in the lab is Nile red. It is a dye that exhibits solvatochromism; its absorption band varies in spectral position, shape and intensity with the nature of its solvent environment, it will fluoresce intensely red in polar environment and blue shift with the changing polarity of its solvent. This makes it ideal for the detection of lipid bodies within Mycobacterium spp. This is because mycobacterial lipid bodies' primary constituents are nonpolar lipids such as triacylglycerols but bacterial cell membranes are primarily polar lipid species. In this chapter we describe an optimal method for using Nile red to distinguish lipid containing (Lipid rich or LR cells) from those without lipid bodies (Lipid Poor or LP). As part of the process we have optimized a method for separating LP and LR cells that does not require the use of an ultracentrifuge or complex separation media. We believe that these methods will facilitate further research in these enigmatic, transient and important cell states.

Notalgia Paresthetica: A Novel Approach to Treatment with Cryolipolysis.

Notalgia paresthetica, a neurosensory syndrome that typically occurs on the upper back, has multiple clinical symptoms with variable degrees of expression in each individual afflicted with the condition. The involved site is usually hyperpigmented and is associated with burning, coldness, hypoesthesia, increased pain, pruritus and/or tingling. In the affected area, the number of nerve fibers may be increased and the cutaneous sensory nerves are altered secondary to localized impingement, central injury, or both. Although multiple therapeutic approaches for notalgia paresthetica have been described, none specifically address the essential pathogenesis of the condition—the altered cutaneous nerves. Cryolipolysis is a well-tolerated nonsurgical technique to reduce the subcutaneous fat layer. Selective apoptosis of adipocytes occurs since the lipid-rich fat cells are more susceptible to cold injury than the surrounding water-rich cells. Not only a marked decrease in pain sensitivity but also a sustained reduction in the density of myelinated and unmyelinated cutaneous nerves has been observed in cryolipolysis-treated skin. Therefore, cryolipolysis is a logical approach to the treatment of notalgia paresthetica. One or more cryolipolysis treatments may be necessary for complete or partial resolution of the individual’s notalgia paresthetica-related cutaneous symptoms. In conclusion, evaluation of cryolipolysis as a noninvasive treatment of patients with notalgia paresthetica is warranted.

Immunomodulatory liposomes targeting liver macrophages arrest progression of nonalcoholic steatohepatitis

ObjectiveNon-alcoholic fatty liver disease (NAFLD) is characterized by hepatic macrophage inflammation, steatosis and fibrosis. Liposomes injected intravenously passively target hepatic myeloid cells and have potential to deliver immunomodulatory compounds and treat disease. We investigated targeting, delivery, immunomodulation and efficacy of liposomes in mice with diet-induced NASH. MethodsLiposome-encapsulated lipophilic curcumin or 1,25-dihydroxy-vitamin D3 (calcitriol) were injected intravenously into mice with diet-induced NASH. Liver and cell liposome uptake was assessed by in vivo imaging and flow cytometry. Immunomodulation of targeted cells were assessed by RNA transcriptome sequencing. NASH was assessed by histological scoring, serum liver enzymes and fasting glucose/insulin and liver RNA transcriptome sequencing. ResultsLiposomes targeted lipid containing MHC class-II+ hepatic dendritic cells in mice and humans. Delivery of liposomal curcumin to hepatic dendritic cells shifted their inflammatory profile towards a regulatory phenotype. Delivery of liposomal curcumin or calcitriol to mice with diet-induced NASH led to reduced liver inflammation, fibrosis and fat accumulation, and reduced insulin resistance. RNA transcriptome sequencing of liver from treated mice identified suppression of pathways of immune activation, cell cycle and collagen deposition. ConclusionsLiposomes are a new strategy to target lipid rich inflammatory dendritic cells and have potential to deliver immunomodulatory compounds to treat NASH.

Label-free optical vibrational spectroscopy to detect the metabolic state of M. tuberculosis cells at the site of disease

Tuberculosis relapse is a barrier to shorter treatment. It is thought that lipid rich cells, phenotypically resistant to antibiotics, may play a major role. Most studies investigating relapse use sputum samples although tissue bacteria may play an important role. We developed a non-destructive, label-free technique combining wavelength modulated Raman (WMR) spectroscopy and fluorescence detection (Nile Red staining) to interrogate Mycobacterium tuberculosis cell state. This approach could differentiate single “dormant” (lipid rich, LR) and “non-dormant” (lipid poor, LP) cells with high sensitivity and specificity. We applied this to experimentally infected guinea pig lung sections and were able to distinguish both cell types showing that the LR phenotype dominates in infected tissue. Both in-vitro and ex-vivo spectra correlated well, showing for the first time that Mycobacterium tuberculosis, likely to be phenotypically resistant to antibiotics, are present in large numbers in tissue. This is an important step in understanding the pathology of relapse supporting the idea that they may be caused by M. tuberculosis cells with lipid inclusions.

Microbial cell disruption for improving lipid recovery using pressurized CO2 : Role of CO2 solubility in cell suspension, sugar broth, and spent media.

The study of in situ gas explosion to lyse the triglyceride-rich cells involves the solubilization of gas (e.g., carbon dioxide, CO2 ) in lipid-rich cells under pressure followed by a rapid decompression, which allows the gas inside the cell to rapidly expand and rupture the cell from inside out. The aim of this study was to perform the cell disruption using pressurized CO2 as well as to determine the solubility of CO2 in Rhodotorula glutinis cell suspension, sugar broth media, and spent media. Cell disruption of R. glutinis was performed at two pressures of 2,000 and 3,500 kPa, respectively, at 295.2 K, and it was found from both scanning electron microscopy (SEM) and plate count that a substantial amount of R. glutinis was disrupted due to the pressurized CO2 . We also found a considerable portion of lipid present in the aqueous phase after the disruption at P = 3,500 kPa compared to control (no pressure) and P = 2,000 kPa, which implied that more intracellular lipid was released due to the pressurized CO2 . Solubility of CO2 in R. glutinis cell suspension was found to be higher than the solubility of CO2 in both sugar broth media and spent media. Experimental solubility was correlated using the extended Henry's law, which showed a good agreement with the experimental data. Enthalpy and entropy of dissolution of CO2 were found to be -14.22 kJmol-1 and 48.10 kJmol-1 K-1 , 9.64 kJmol-1 and 32.52 kJmol-1 K-1 , and 7.50 kJmol-1 and 25.22 kJmol-1 K-1 in R. glutinis, spent media, and sugar broth media, respectively. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:737-748, 2017.

Intracoronary Imaging in the Detection of Vulnerable Plaques

Coronary artery disease is the result of atherosclerotic changes to the coronary arterial wall, comprising endothelial dysfunction, vascular inflammation and deposition of lipid-rich macrophage foam cells. Certain high-risk atherosclerotic plaques are vulnerable to disruption, leading to rupture, thrombosis and the clinical sequelae of acute coronary syndrome. Though recognised as the gold standard for evaluating the presence, distribution and severity of atherosclerotic lesions, invasive coronary angiography is incapable of identifying non-stenotic, vulnerable plaques that are responsible for adverse cardiovascular events. The recognition of such limitations has impelled the development of intracoronary imaging technologies, including intravascular ultrasound, optical coherence tomography and near-infrared spectroscopy, which enable the detailed evaluation of the coronary wall and atherosclerotic plaques in clinical practice. This review discusses the present status of invasive imaging technologies; summarises up-to-date, evidence-based clinical guidelines; and addresses questions that remain unanswered with regard to the future of intracoronary plaque imaging.

Maximising the use of freshly isolated human hepatocytes

IntroductionFreshly isolated human hepatocytes are the best model for predicting adverse drug reactions. However, their preparation and use present the investigator with many variables that are beyond their control. These include operation continuity and timing, size and number of cut surfaces on liver tissue and the prior history of the patient. To exploit the potential of freshly isolated human hepatocytes a method is required to preserve the cells in their initial in vivo like state. This experimental pausing allows experiments to be prioritised at convenient times of the day. MethodsA novel approach for selecting viable human hepatocytes by functional attachment to a gelatin gel is described rather than relying on their physical characteristics. The cells are preserved as a monolayer on the semi-solid support at 10°C as single spherical entities. ResultsThe hepatocytes can be released into suspension, when required, by a temperature transition to 37°C for 20min. The cells can be used in suspension or as a monolayer. The length of preservation depends upon the source tissue. Hepatocytes from normal liver can be maintained for at least 4days and demonstrated to have the same level of CYP3A4 and the enzymes involved in glucuronidation and sulphation as freshly isolated cells. Cells from fatty liver, attached to gelatin, vary in their preservation time but it is at least 24h and so confluent monolayers, that survive at 37°C can be generated the following day. DiscussionThe technique enables freshly isolated human hepatocytes to be used more effectively. They can be preserved in times of plenty so more experimentation is possible. Alternatively, with poorer fatty cells the initial attachment on gelatin enables confluent monolayers of lipid rich cells to be studied.

Novel somatic mutations in primary hyperaldosteronism are related to the clinical, radiological and pathological phenotype.

Aldosterone-producing adenomas (APAs) and bilateral adrenal hyperplasia are important causes of secondary hypertension. Somatic mutations in KCNJ5, CACNA1D, ATP1A1, ATP2B3 and CTNNB1 have been described in APAs. To characterize clinical-pathological features in APAs and unilateral adrenal hyperplasia, and correlate them with genotypes. Retrospective study. Clinical and pathological characteristics of 90 APAs and seven diffusely or focally hyperplastic adrenal glands were reviewed, and samples were examined for mutations in known disease genes by Sanger or exome sequencing. Mutation frequencies were as follows: KCNJ5, 37·1%; CACNA1D, 10·3%; ATP1A1, 8·2%; ATP2B3, 3·1%; and CTNNB1, 2·1%. Previously unidentified mutations included I157K, F154C and two insertions (I150_G151insM and I144_E145insAI) in KCNJ5, all close to the selectivity filter, V426G_V427Q_A428_L433del in ATP2B3 and A39Efs*3 in CTNNB1. Mutations in KCNJ5 were associated with female and other mutations with male gender (P = 0·007). On computed tomography, KCNJ5-mutant tumours displayed significantly greater diameter (P = 0·023), calculated area (P = 0·002) and lower precontrast Hounsfield units (P = 0·0002) vs tumours with mutations in other genes. Accordingly, KCNJ5-mutant tumours were predominantly comprised of lipid-rich fasciculata-like clear cells, whereas other tumours were heterogeneous (P = 5 × 10(-6) vs non-KCNJ5 mutant and P = 0·0003 vs wild-type tumours, respectively). CACNA1D mutations were present in two samples with hyperplasia without adenoma. KCNJ5-mutant tumours appear to be associated with fasciculata-like clear cell predominant histology and tend to be larger with a characteristic imaging phenotype. Novel somatic KCNJ5 variants likely cause adenomas by loss of potassium selectivity, similar to previously described mutations.

The 10 Hounsfield units unenhanced computed tomography attenuation threshold does not apply to cortisol secreting adrenocortical adenomas.

Computed tomography (CT) unenhanced attenuation value of <10 Hounsfield units (HU) has an excellent specificity (98%) to diagnose lipid-rich adrenocortical adenomas (ACAs) with a weaker sensitivity (71%). To determine from a routine clinical perspective if unenhanced attenuation value is influenced by cortisol secretion in ACAs. This was a retrospective study of cases collected between 2009 and 2012. This study was conducted in a tertiary-care university hospital. Seventy-two patients operated on for an ACA (Weiss score ≤ 2) were analysed. Thirty-four patients had an ACA oversecreting cortisol (Cush-ACA). Thirty-eight patients had an ACA without cortisol oversecretion (Non Hyper-ACA). CT unenhanced attenuation value was correlated with the functional status. The Weiss score items were analysed. Among the 34 patients with a Cush-ACA a minority (n = 7) had an unenhanced attenuation value under 10 HU. Among the high precontrast density (> 10 HU) Cush-ACAs, washout analysis after contrast administration was consistent with the benign nature of the tumor in ∼ 60% of the cases. Less than 25% clear cells (lipid-rich cells), a Weiss score item, was present in 50% of the Cush-ACAs in favour of a lipid-poor content. Unenhanced attenuation value has a poor sensitivity to diagnose an ACA in case of cortisol oversecretion due to poor lipid content. Nevertheless, the accuracy of washout analysis was preserved in the group of Cush-ACAs.

Phenotypic resistance in mycobacteria: is it because I am old or fat that I resist you?

We aimed to explore the phenomenon of phenotypic resistance to antimycobacterial antibiotics and to determine whether this was associated with cell age or the presence of lipid bodies. The accumulation of lipid-body-positive [lipid-rich (LR)] cells was followed using cell staining and flow cytometry. LR cells of Mycobacterium smegmatis, Mycobacterium marinum, Mycobacterium fortuitum and Mycobacterium bovis (BCG) were separated from non-lipid-body-containing [lipid-poor (LP)] cells and their MBCs determined. We also compared the MBCs for LR and LP cells from 'old' and 'young' cultures. The LR cells of all species were more resistant to antibiotics than LP cells. For BCG, the susceptibility ratios were as follows: rifampicin, 5×; isoniazid, 16.7×; ethambutol, 5×; and ciprofloxacin, 5×. Phenotypic resistance was found in LR cells irrespective of cell age. We have shown that phenotypic antibiotic resistance is associated with the presence of lipid bodies irrespective of cell age. These data have important implications for our understanding of relapse in mycobacterial infections.

Dynamics of the lipid droplet proteome of the Oleaginous yeast rhodosporidium toruloides.

Lipid droplets (LDs) are ubiquitous organelles that serve as a neutral lipid reservoir and a hub for lipid metabolism. Manipulating LD formation, evolution, and mobilization in oleaginous species may lead to the production of fatty acid-derived biofuels and chemicals. However, key factors regulating LD dynamics remain poorly characterized. Here we purified the LDs and identified LD-associated proteins from cells of the lipid-producing yeast Rhodosporidium toruloides cultured under nutrient-rich, nitrogen-limited, and phosphorus-limited conditions. The LD proteome consisted of 226 proteins, many of which are involved in lipid metabolism and LD formation and evolution. Further analysis of our previous comparative transcriptome and proteome data sets indicated that the transcription level of 85 genes and protein abundance of 77 proteins changed under nutrient-limited conditions. Such changes were highly relevant to lipid accumulation and partially confirmed by reverse transcription-quantitative PCR. We demonstrated that the major LD structure protein Ldp1 is an LD marker protein being upregulated in lipid-rich cells. When overexpressed in Saccharomyces cerevisiae, Ldp1 localized on the LD surface and facilitated giant LD formation, suggesting that Ldp1 plays an important role in controlling LD dynamics. Our results significantly advance the understanding of the molecular basis of lipid overproduction and storage in oleaginous yeasts and will be valuable for the development of superior lipid producers.

Rapid method to screen and sort lipid accumulating microalgae

The present work established an efficient staining method for fluorescence activated cell sorting (FACS) with Chlorococcum littorale maintaining cellular viability. The method was designed to detect high-lipid cells and to guarantee cellular viability. BODIPY505/515 (BP) was more suitable to FACS when compared to Nile red. The optimum concentrations were 0.4μgml−1 of BP, 0.1% DMSO or 0.35% ethanol. Both ethanol and DMSO were equally efficient and assured cellular viability after the staining and sorting. Here a method is presented to rapidly screen and sort lipid rich cells of C. littorale with FACS, which can be used to produce new inoculum with increased cellular lipid content.

Monocyte recruitment and macrophage proliferation in atherosclerosis

Copyright © Polskie Towarzystwo Kardiologiczne INTRODUCTION In the famous ‘Atlas of atherosclerosis: progression and regression’, Herbert Stary [1] outlines in remarkable detail the development of human atherosclerosis. The photographs depicting lesions at all stages of development, from newborns to the elderly, capture in stunning detail the complexity of an evolving disease. Type I lesions develop, Stary explains, at sites of non-laminar flow and low shear stress, which disturbs endothelial function. Type II lesions are defined by macrophage foam cells, and type III preatheromas feature small pools of extracellular lipids. As the disease worsens, an easily discernible core of extracellular lipid marks a type IV atheroma, fibrous thickening corresponds to a type V atheroma, the appearance of fissures, haematoma, and thrombi denote type VI atheroma, and finally, calcification is a type VII complicated atheroma [1]. Each of these histological observations reflect a biological process; each has its corresponding cluster of scientists and clinicians devoted to understanding how it can be harnessed to prevent or treat disease. As the underlying pathology causing most myocardial infarctions and strokes, atherosclerosis is, after all, the deadliest disease in the world [2]. Among the many mechanisms that contribute to the development and complications of atherosclerosis, macrophage accumulation occurs early and persists throughout most of a lesion’s evolution. The best evidence that macrophages are functionally important — rather than simply markers — can be found in animal studies. The most widely-used murine models of atherosclerosis, the Apoe–/– and Ldlr–/– mice, though far from perfect reflections of human disease, are nevertheless fair approximations to show how lesions develop. In both models, macrophages are prominent in early and advanced lesions, where they ingest oxidised lipoproteins via scavenger receptors and, as lipid-rich foam cells, become part of the disease’s physical bulk [3]. Although many of the functions by which macrophages influence atherosclerosis have been deciphered, their ontogeny has continued to perplex. On the one extreme, lesional macrophages may be developing from resident precursors or stem cells through local differentiation and proliferation, requiring no input from the circulation. At the other extreme, macrophage accumulation may be the exclusive consequence of replenishment from blood monocytes, requiring no input from resident cells. Identifying the mechanisms can be therapeutically relevant. If macrophages are harmful to disease and originate only from local precursors, then approaches targeting the vessel wall and its environment should be explored. If, however, macrophages originate exclusively from blood monocytes, then targeting environments where monocytes arise, such as the bone marrow or spleen, may be the more effective strategy.

Abstract P6-12-16: Treatment with indol-3-carbinol (I3C) in a mice xenograft model of canine breast carcinoma alter hormone metabolism and produce liver metastases

Abstract Indol-3-carbinol (I3C) is a chemical component found mainly in the cruciferous family lants (i.e. broccoli, cabbage, cauliflower, etc.). I3C is considered a potential anticancer agent that is naturally present in the diet and could prevents the development of certain tumors, activating tumor suppressor genes, genes involved in apoptosis and detoxifying genes. It has been observed in vitro that suppresses cell proliferation and induces apoptosis in some breast cancers. I3C is also estrogen receptor (ER) and androgen receptor (AR)Antagonist. Therefore, studies using I3C in breast cancer animal models for its action as a preventive or therapeutic substance have started. There are no data on the influence of I3C in human and/or spontaneous or experimental canine inflammatory breast cancers, that has shown to have distinct pathogenic mechanisms. The aim of this study was to analyze the effect of I3C administration at a comparable dose used as preventive in humans in a xenograft model of canine inflammatory carcinoma established in SCID mice. Eighteen mice were used as controls and 13 mice were treated with I3C dose 150 mg/kg/day. Tumor cell proliferation was decreased in the I3C-treated xenografts (lower Ki-67 index, p &amp;lt;0.001), but neoplasms were more frequently ulcerated (p = 0.028), and mice had a higher number of emboli in dermis (p = 0.012) (characteristic of inflammatory breast carcinoma) and liver metastases (p = 0.05). In the treated group the presence of a subpopulation of neoplastic lipid-rich cells was characteristic (p = 0.001) togetehr with a higher content of intratumor androgens (p = 0.02) and serum steroid hormone (p &amp;lt;0 05). These results indicate that treatment with I3C, as administered, is not suitable for inflammatory breast carcinoma. This study was funded by the research project of the Ministry of Science and Innovation SAF2009-10572. Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P6-12-16.