Inhibition of Pro-inflammatory and Anti-apoptotic Biomarkers during Experimental Oral Cancer Chemoprevention by Dietary Black Raspberries

Abstract

Oral cancer continues to be a significant public health problem worldwide. Recently conducted clinical trials demonstrate the ability of black raspberries (BRBs) to modulate biomarkers of molecular efficacy that supports a chemopreventive strategy against oral cancer. However, it is essential that a preclinical animal model of black raspberry (BRB) chemoprevention which recapitulates human oral carcinogenesis be developed, so that we can validate biomarkers and evaluate potential mechanisms of action. We therefore established the ability of BRBs to inhibit oral lesion formation in a carcinogen-induced rat oral cancer model and examined potential mechanisms. F344 rats were administered 4-nitroquinoline 1-oxide (4NQO) (20 µg/ml) in drinking water for 14 weeks followed by regular drinking water for 6 weeks. At week 14, rats were fed a diet containing either 5 or 10% BRB, or 0.4% ellagic acid (EA), a BRB phytochemical. Dietary administration of 5 and 10% BRB reduced oral lesion incidence and multiplicity by 39.3 and 28.6%, respectively. Histopathological analyses demonstrate the ability of BRBs and, to a lesser extent EA, to inhibit the progression of oral cancer. Oral lesion inhibition by BRBs was associated with a reduction in the mRNA expression of pro-inflammatory biomarkers Cxcl1, Mif, and Nfe2l2 as well as the anti-apoptotic and cell cycle associated markers Birc5, Aurka, Ccna1, and Ccna2. Cellular proliferation (Ki-67 staining) in tongue lesions was inhibited by BRBs and EA. Our study demonstrates that, in the rat 4NQO oral cancer model, dietary administration of BRBs inhibits oral carcinogenesis via inhibition of pro-inflammatory and anti-apoptotic pathways.

Characterization of the Functional Changes in Mouse Gut Microbiome Associated with Increased Akkermansia muciniphila Population Modulated by Dietary Black Raspberries

“…A total of 20 specific-pathogen free C57BL/6 mice (∼8 week old), purchased from Jackson Laboratories, were housed in the animal facility of the University of Georgia for 1 week to acclimate with standard pelleted rodent diet and tap water ad libitum provided (protocol number: A2013 06-033-Y3-A3). The BRB diet was prepared essentially as described in ref ( 52 ). Briefly, whole ripe BRBs of the Jewel variety were freeze-dried and ground into powder.…”

Combination of IFITM1 knockdown and radiotherapy inhibits the growth of oral cancer

“…This amounts to more than one fatality every hour due to oral cancer. Worldwide, approximately 300 000 cases and 145 000 deaths are reported annually . Research dedicated to radiotherapy tolerance is on the rise, and the present research seeks to contribute to the treatment strategies of oral neoplasm using radiotherapy in combination with the inhibition of IFITM1.…”

Immune Suppression Mediated by STAT4 Deficiency Promotes Lymphatic Metastasis in HNSCC

“…RNA was extracted using Direct-zol RNA Miniprep kit (Zymo Research, Irvine, CA, USA) and reverse transcribed to cDNA using the High Capacity cDNA Reverse Transcription Kit (Applied Biosystems, Foster City, CA, USA). Primer sequences for PCR were created using the IDT RealTime qPCR Tool (https://www.idtdna.com/scitools/ Applications/RealTimePCR/, Integrated DNA Technologies, Coralville, IA, USA) (26,27). Real time PCR of cDNA samples was performed using the PowerUp SYBR Green Master Mix (Thermofisher Scientific, Foster City, CA, USA) with beta actin (Actb) as a reference gene.…”