Inhibition of the PI3K/AKT signaling pathway increases efficacy of doxorubicin and its derivative AD198 in bladder and oral cancers

Doxorubicin (Dox) is a successful chemotherapy to treat various cancers, including bladder and oral cancers. Many patients initially respond to Dox-based regimens, however often cancers become resistant. A novel derivatives of Dox, e.g. N-benzyladriamycin-14-valerate (AD198), have been developed to overcome Dox-induced drug resistance and cardiotoxicity. The purpose of this thesis was to determine the efficacy of AD198 and Dox in bladder and oral cancers in vitro. Part-I of this dissertation focuses on the bladder cancer, including discussing risk factors, diagnosis, staging, and current treatment options, following by a description of altered molecular mechanisms responsible cancer progression. This section also focuses on alternative experimental drugs and current clinical trials designed to target specific molecular markers of bladder cancer. Part-II of this dissertation compares the efficacy of AD198 and Dox and its molecular mechanisms of action in human T24 and UMUC3 bladder cells in vitro. AD198 was more effective than Dox in inhibition of cell viability of T24 and UMUC3 cells. Both Dox and AD198 significantly induced apoptosis in caspase-dependent and -independent manners. Dox and AD198 activated the pro-apoptotic p38 MAPK pathway; however, they also increased phosphorylation of AKT, a pro-survival signaling pathway, in T24 and UMUC3 cells. Combined treatment of PI3K inhibitor (LY294002) with Dox or AD198 inhibited cell viability of T24 and UMUC3 cells more effectively than any drug treatment alone. Part-III of this dissertation discusses oral cancer, with special focus on causes, diagnosis, treatment, molecular pathogenesis, and potential molecular targets for treatments. Part IV of this dissertation focuses on evaluation of the efficacy of Dox and its novel derivative AD198 in human (SCC25 and 1483), canine K9OSCC-Abby, and feline (FeOSCC-Sidney) oral squamous cell carcinoma cells in vitro. Dox and AD198 had a better anti-proliferative effect than Dox in human and canine OSCC. Our results suggest that the combined therapy of an anthracycline compound with inhibitor of PI3K/AKT pathway is a more effective treatment. Part V of this dissertation discusses the implications of these studies and examines current literature on the potential of targeting PI3K/AKT to increase the efficacy of anthracycline treatments in bladder and oral cancers

Comparison of Content in Phenolic Compounds and Antioxidant Capacity in Grains of White, Red, and Black Sorghum Varieties Grown in the Mediterranean Area

orghum (Sorghum bicolor L.) is a gluten-free cereal with many varieties containing greater amounts of bioactive compounds than other grains. In this work, the phenolic composition and antioxidant capacity were compared in three sorghum varieties grown in the Mediterranean, consisting of different pericarp colors: white, red, and black. The qualitative and quantitative profiles of acidified aqueous methanol extracts of whole grain sorghum flours were elucidated through ultrahigh performance liquid chromatography (UHPLC) coupled to photodiode array (PDA) and ion trap time-of-flight (IT-TOF) mass spectrometry. The investigated sorghum varieties contained polyamines, such as spermidines, and polyphenolic compounds belonging to different classes: hydroxycinnamic acids, flavanols, flavones, and flavanones. Pyrano-flavanone-flavanols were detected only in the black sorghum. Precolumn and postcolumn 2,2′-diphenyl-1-picrylhydrazyl (DPPH)· radical scavenging assays to determine the main antioxidant compounds isolated from sorghum were developed. These results will further help develop sorghum varieties containing polyphenols relevant to the potential prevention of human diseases

A Comparative Study on Phenolic Content, Antioxidant Activity and Anti-Inflammatory Capacity of Aqueous and Ethanolic Extracts of Sorghum in Lipopolysaccharide-Induced RAW 264.7 Macrophages

Sorghum is an important cereal with diverse phenolic compounds that have potential health promoting benefits. The current study comparatively characterized the phenolic contents of two novel black-seeded sorghum lines (SC84 and PI570481) using different extraction systems (water, ethanol and their acidified counterparts) and evaluated their antioxidant and anti-inflammatory activities. Phenolic compositions were determined by spectrophotometric assays and HPLC analysis. Antioxidant activities were assessed by radical scavenging effects on nitric oxide (NO) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radicals, and the oxygen radical absorbance capacity (ORAC). Anti-inflammatory capacity was estimated by measuring levels of pro-inflammatory markers produced by lipopolysaccharide (LPS)-induced RAW 264.7 macrophages. Results showed that effects of solvent types and HCl on extraction efficiency differed among phenolic compounds and sorghum samples. Tannins were the most dominant polyphenols in the studied extracts (11.11–136.11 mg epicatechin equivalent/g sorghum). Sorghum extracts exerted more potent scavenging activity on DPPH than NO radicals. In LPS-activated RAW 264.7 cells, sorghum extracts dose-dependently inhibited the production of NO, interleukin-6 (IL-6), and intracellular reactive oxygen species (ROS), with ethanolic extracts showing greater anti-inflammatory activity. Positive correlations were noted between tannin content and DPPH radical scavenging activity, and anti-inflammatory capacity. These results suggest the potential role of tannin-rich sorghum extracts against inflammation and associated diseases

Anti-Adipogenic Activity of High-Phenolic Sorghum Brans in Pre-Adipocytes

Obesity is one of the leading public health problems that can result in life-threatening metabolic and chronic diseases such as cardiovascular diseases, diabetes, and cancer. Sorghum (Sorghum bicolor (L.) Moench) is the fifth most important cereal crop in the world and certain genotypes of sorghum have high polyphenol content. PI570481, SC84, and commercially available sumac sorghum are high-polyphenol genotypes that have demonstrated strong anti-cancer activities in previous studies. The objective of this study was to explore a potential anti-obesity use of extracts from sorghum bran in the differentiation of 3T3-L1 preadipocytes and to investigate cellular and molecular responses in differentiated adipocytes to elucidate related mechanisms. None of the four different sorghum bran extracts (PI570481, SC84, Sumac, and white sorghum as a low-polyphenol control) caused cytotoxicity in undifferentiated and differentiated 3T3-L1 cells at doses used in this study. Sorghum bran extracts (PI570481, SC84, and Sumac) reduced intracellular lipid accumulation and expression of adipogenic and lipogenic proteins in a dose-dependent manner in differentiated 3T3-L1 cells. The same polyphenol containing sorghum bran extracts also repressed production of reactive oxygen species (ROS) and MAPK signaling pathways and repressed insulin signaling and glucose uptake in differentiated 3T3-L1 cells. These data propose a potential use of high-phenolic sorghum bran for the prevention of obesity.https://doi.org/10.3390/nu1407149

Polyphenol Containing Sorghum Brans Exhibit an Anti-Cancer Effect in Apc Min/+ Mice Treated with Dextran Sodium Sulfate

Colon cancer (CC) is considered a high-risk cancer in developed countries. Its etiology is correlated with a high consumption of red meat and low consumption of plant-based foods, including whole grains. Sorghum bran is rich in polyphenols. This study aimed to determine whether different high-phenolic sorghum brans suppress tumor formation in a genetic CC rodent model and elucidate mechanisms. Tissue culture experiments used colorectal cancer cell lines SW480, HCT-116 and Caco-2 and measured protein expression, and protein activity. The animal model used in this study was APC Min+/mouse model combined with dextram sodium sulfate. High phenolic sorghum bran extract treatment resulted in the inhibition of proliferation and induced apoptosis in CC cell lines. Treatment with high phenolic sorghum bran extracts repressed TNF-α-stimulated NF-κB transactivation and IGF-1-stimulated PI3K/AKT pathway via the downregulation of β-catenin transactivation. Furthermore, high-phenolic sorghum bran extracts activated AMPK and autophagy. Feeding with high-phenolic sorghum bran for 6 weeks significantly suppressed tumor formation in an APC Min/+ dextran sodium sulfate promoted CC mouse model. Our data demonstrates the potential application of high-phenolic sorghum bran as a functional food for the prevention of CC

Antimicrobial Activity of Sorghum Phenolic Extract on Bovine Foodborne and Mastitis-Causing Pathogens

Antimicrobial resistance in bacterial pathogens associated with bovine mastitis and human foodborne illnesses from contaminated food and water have an impact on animal and human health. Phenolic compounds have antimicrobial properties and some specialty sorghum grains are high in phenolic compounds, and the grain extract may have the potential as a natural antimicrobial alternative. The study’s objective was to determine antimicrobial effects of sorghum phenolic extract on bacterial pathogens that cause bovine mastitis and human foodborne illnesses. Bacterial pathogens tested included Escherichia coli, Salmonella Typhimurium, Campylobacter jejuni, Campylobacter coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Klebsiella oxytoca, Staphylococcus aureus, and Enterococcus faecalis. Antibacterial activities of sorghum phenolic extracts were determined by agar-well diffusion assay. Sorghum phenolic extract was added to the wells in concentrations of 0, 100, 200, 500, 1000, or 4000 µg/mL. The control wells did not receive phenolic extract. Plates were incubated for 18–24 h, and the diameter of each zone of inhibition was measured. The results indicated that sorghum phenolic extract had inhibitory effects on Staphylococcus aureus, Enterococcus faecalis, Campylobacter jejuni, and Campylobacter coli

Chemical Composition, Fatty Acid and Mineral Content of Food-Grade White, Red and Black Sorghum Varieties Grown in the Mediterranean Environment

none15siGrain sorghum (Sorghum bicolor) is a gluten-free cereal grown around the world and is a food staple in semi-arid and subtropical regions. Sorghum is a diverse crop with a range of pericarp colour including white, various shades of red, and black, all of which show health-promoting properties as they are rich sources of antioxidants such as polyphenols, carotenoids, as well as micro- and macro-nutrients. This work examined the grain composition of three sorghum varieties possessing a range of pericarp colours (white, red, and black) grown in the Mediterranean region. To determine the nutritional quality independent of the contributions of phenolics, mineral and fatty acid content and composition were measured. Minor differences in both protein and carbohydrate were observed among varieties, and a higher fibre content was found in both the red and black varieties. A higher amount of total saturated fats was found in the white variety, while the black variety had a lower amount of total unsaturated and polyunsaturated fats than either the white or red varieties. Oleic, linoleic, and palmitic were the primary fatty acids in all three analysed sorghum varieties. Significant differences in mineral content were found among the samples with a greater amount of Mg, K, Al, Mn, Fe, Ni, Zn, Pb and U in both red and black than the white sorghum variety. The results show that sorghum whole grain flour made from grain with varying pericarp colours contains unique nutritional properties.Pontieri, Paola; Troisi, Jacopo; Calcagnile, Matteo; Bean, Scott R; Tilley, Michael; Aramouni, Fadi; Boffa, Antonio; Pepe, Giacomo; Campiglia, Pietro; Del Giudice, Fabio; Chessa, Alberto L; Smolensky, Dmitriy; Aletta, Mariarosaria; Alifano, Pietro; Del Giudice, LuigiPontieri, Paola; Troisi, Jacopo; Calcagnile, Matteo; Bean, Scott R; Tilley, Michael; Aramouni, Fadi; Boffa, Antonio; Pepe, Giacomo; Campiglia, Pietro; Del Giudice, Fabio; Chessa, Alberto L; Smolensky, Dmitriy; Aletta, Mariarosaria; Alifano, Pietro; Del Giudice, Luig