S-allylcysteine, a garlic constituent, inhibits 7,12-dimethylbenz[a]anthracene-induced hamster buccal pouch carcinogenesis

The effect of S-allylcysteine (SAC), a water-soluble garlic constituent, on 7,12-dimethylbenz[a]anthracene (DMBA)-induced hamster buccal pouch (HBP) carcinogenesis was investigated in male Syrian hamstes. Forty hamsters were divided into 4 groups of 10 animals. The right buccal pouches of the animals in Group I were painted with a 0.5% solution of DMBA in liquid paraffin three times a week. The animals in Group II were painted with DMBA as in Group I and, in addition, received 200 mg/kg body wt p.o. SAC three times a week on days alternate to DMBA application. Group III animals received SAC as in Group II. Group IV animals received neither DMBA nor SAC and served as the control. The hamsters were killed after an experimental period of 14 wk. Measurement of lipid peroxidation, the antioxidant enzymes superoxide dismutase (SOD) and catalase, in the buccal pouch mucosa, liver, and circulation was used to monitor the chemopreventive potential of SAC. All hamsters painted with DMBA alone developed tumors identified histologically as well-differentiated squamous cell carcinomas. In hamsters bearing DMBA-induced buccal pouch tumors, diminished lipid peroxidation in the tumor tissue was accompanied by decreased activities of SOD and catalase, whereas in the liver and circulation, enhanced lipid peroxidation was associated with compromised antioxidant defenses. Administration of SAC suppressed the incidence of DMBA-induced HBP tumors as revealed by the absence of carcinomas. Histologically, only keratosis was observed. SAC modulated DMBA-induced decreased susceptibility of the HBP to lipid peroxidation while simultaneously enhancing SOD and catalase activities, whereas in the liver and circulation, SAC decreased the extent of lipid peroxidation and significantly enhanced antioxidant activities. We suggest that SAC exerts its chemopreventive effects by modulating lipid peroxidation and enhancing antioxidant activities in the target organ as well as in the liver and circulation.     

Correlation of xenobiotic-metabolizing enzymes,oxidative stress and NFκB signaling with histological grade and menopausal status in patients with adenocarcinoma of the breast

BackgroundAdenocarcinoma of the breast is the most common cancer worldwide and accounts for the highest morbidity and mortality. The increasing global incidence of breast cancer emphasizes the need to understand the molecular mechanisms of breast tumorigenesis. The present study was designed to correlate changes in xenobiotic-metabolizing enzymes (XME), oxidative stress and NFκB signaling with histological grading and menopausal status in breast cancer patients.MethodSixty breast cancer patients histologically categorized as grades I, II and III, and as pre- and postmenopausal were chosen for the study. We analyzed phase I and phase II XME activities as well as the expression of the CYP isoforms CYP1A1 and CYP1B1, oxidative stress markers, and the expression of NFκB family members in tumor and adjacent tissues by immunohistochemical localization and Western blot analyses.ResultsThe breast tumors analyzed in the present study were characterized by increased activities of xenobiotic-metabolizing enzymes and enhanced oxidative damage to lipids, proteins, and DNA associated with variations in the expression of NFκB family members. The magnitude of the changes was however more pronounced in premenopausal patients and in grade III breast tumors.ConclusionThe present study delineates the correlation between XME-mediated oxidative stress and NFκB signaling that leads to the development of breast cancer.     

Fractional allele loss indicates distinct genetic populations in the development of squamous cell carcinoma of the head and neck (SCCHN)

Loss of heterozygosity (LOH) had been widely used to assess genetic instability in tumours and a high LOH on chromosome arms 3p, 9p and 17p has been considered to be a common event in squamous cell carcinoma of the head and neck (SCCHN). We have investigated LOH in 52 SCCHN using a range of microsatellite markers. LOH was observed in 69% of individuals on 17p using seven markers, in 64% of individuals on 3p using 17 markers and in 61% of individuals on 9p using 11 markers. Fractional allele loss (FAL) has been calculated for each tumour (FAL is the number of chromosomal arms showing LOH divided by the number of informative chromosomal arms) and a median FAL value of 0.25 was obtained in the 52 SCCHN studied. The LOH data were examined on the basis of FAL scores: low FAL (LFAL), 0.00-0.19; medium FAL (MFAL), 0.20-0.32; high FAL (HFAL), 0.33-0.88. HFAL tumours demonstrated a significantly higher LOH on chromosome arms 3p, 9p and 17p, with 94% LOH on 3p, 94% on 9p and 100% on 17p compared with LFAL tumours. Six of the 16 patients in the LFAL group were found to have no LOH on 3p, 9p or 17p and of these four had LOH at other sites, on chromosomes 2p25-p24, 5q21-22, 7pter-p22, 8q13-q22.1, 11q23.3, 13q32, 17q, 18p11.21, 18q21.31 and 19q12-q13.1. These results indicate that LFAL patients form a subset of SCCHN tumours with distinct molecular initiating events which may represent a discrete genetic population.     

Altered cytokeratin expression during chemoprevention of experimental hamster buccal pouch carcinogenesis by garlic

BACKGROUND: Cytokeratins (also known as keratins (K)) are members of the family of intermediate filaments and form major components of the mammalian epithelial cell cytoskeleton. Cytokeratins have emerged as reliable cellular markers of oral cancer development and chemoprevention because of their abundance, stability and high antigenicity. METHODS: We investigated the effect of aqueous garlic extract on cytokeratin expression during 7,12-dimethylbenz[a]anthracene (DMBA)-induced hamster buccal pouch (HBP) carcinogenesis. Hamsters were divided into four groups of six animals. Animals in group 1 were painted with a 0.5% solution of DMBA in liquid paraffin, on the right buccal pouches, three times a week for 14 weeks. Group 2 animals were painted with DMBA as in group 1 and also received 250 mg/kg body weight aqueous garlic extract orally on alternate days to the DMBA application. Group 3 animals received garlic extract only, as in group 2. Group 4 animals received neither DMBA nor garlic extract and served as the control. The hamsters were killed after an experimental period of 14 weeks. RESULTS: Cytokeratin expression was studied using human monoclonal antibodies AE1 and AE3, which react with type I and II keratins. In DMBA-induced squamous cell carcinomas, decreased expression of high molecular weight keratins was observed. Administration of garlic extract to animals painted with DMBA suppressed HBP carcinomas and restored normal cytokeratin expression. CONCLUSION: The results of the present study suggest that inhibition of HBP carcinogenesis by garlic may be due to its regulatory effects on differentiation, tumour invasiveness, migratory and metastatic potential. We suggest that one of the mechanisms of tumour inhibition by garlic is an influence on cellular differentiation.     

Combinatorial antigenotoxic and anticarcinogenic effects of tomato and garlic through modulation of xenobiotic-metabolizing enzymes during hamster buccal pouch carcinogenesis

OBJECTIVE: Combination chemoprevention by dietary agents is a promising approach toward cancer control. Many dietary agents are known to prevent experimental mutagenesis and carcinogenesis by modulating xenobiotic-metabolizing enzymes. The present study evaluated the combinatorial chemopreventive effects of tomato and garlic on hamster buccal pouch carcinogenesis induced by 7,12-dimethylbenz[a]anthracene (DMBA). METHODS: Hamsters were assigned to one of four groups. The right buccal pouches of animals in group 1 were painted with 0.5% DMBA three times a week. The right buccal pouches of animals in group 2 were painted with DMBA and received intragastric administration of a combined dose of tomato and garlic on days alternate to DMBA application. Animals in group 3 were given chemopreventive agents alone. Animals in group 4 served as controls. Levels of phase I and II enzymes and the frequency of bone marrow micronuclei were used as biomarkers of chemoprevention. RESULTS: All the hamsters painted with DMBA alone developed buccal pouch carcinomas that exhibited increased activities of xenobiotic-metabolizing enzymes associated with increased frequencies of bone marrow micronuclei. In the liver, an increase in phase I enzymes was accompanied by compromised phase II detoxification activities. Combined administration of tomato and garlic effectively suppressed the incidence and mean tumor burden of hamster buccal pouch carcinomas. In addition, tomato and garlic combination significantly decreased phase I enzymes and increased phase II enzyme activities in the pouch and liver with a decreased incidence of bone marrow micronuclei. CONCLUSION: From these results, we suggest that modulation of xenobiotic-metabolizing enzymes exerted by tomato and garlic combination plays a key role in mitigating the mutagenic and carcinogenic effects of DMBA.     

Lycopene: a review of its potential as an anticancer agent

Dietary chemoprevention has emerged as a cost effective approach to control most prevalent chronic diseases including cancer. In particular, tomato and tomato products are recognised to confer a wide range of health benefits. Epidemiological studies have provided evidence that high consumption of tomatoes effectively lowers the risk of reactive oxygen species (ROS)-mediated diseases such as cardiovascular disease and cancer by improving the antioxidant capacity. Tomatoes are rich sources of lycopene, an antioxidant carotenoid reported to be a more stable and potent singlet oxygen quenching agent compared to other carotenoids. In addition to its antioxidant properties, lycopene shows an array of biological effects including cardioprotective, anti-inflammatory, antimutagenic and anticarcinogenic activities. The anticancer activity of lycopene has been demonstrated both in in vitro and in vivo tumour models. The mechanisms underlying the inhibitory effects of lycopene on carcinogenesis could involve ROS scavenging, upregulation of detoxification systems, interference with cell proliferation, induction of gap-junctional communication, inhibition of cell cycle progression and modulation of signal transduction pathways. This review outlines the sources, structure, absorption, metabolism, bioavailability and pharmacological properties of lycopene with special reference to its antioxidant and anticarcinogenic effects.     

Glycogen synthase kinase-3β mediated regulation of matrix metalloproteinase-9 and its involvement in oral squamous cell carcinoma progression and invasion

Purpose

Oral squamous cell carcinoma (OSCC)-related deaths mainly result from invasion of the tumor cells into local cervical lymph nodes. It has been reported that progressive basement membrane loss promotes the metastatic and invasive capacities of OSCCs. Matrix metalloproteinase-9 (MMP-9) is known to play a central role in tumor progression and invasion. However, the role of MMP-9 in OSCC invasion has so far remained paradoxical and little is known about its regulation. Here, we aimed to assess MMP-9 expression regulation and its activation by glycogen synthase kinase-3β during human OSCC progression and invasion.

Methods

In the present study, 178 human OSCC samples, including 118 fresh samples (18 adjacent normal, 42 noninvasive and 58 invasive tumor samples) and 60 archival human tissue microarray (TMA) tongue cancer samples, were included. mRNA expression, protein expression, MMP-9/-2 activity, protein-protein interaction and Snail, c-Myc, β-catenin and TIMP1 expression were assessed using RT-PCR, immunohistochemistry, Western blotting, co-immunoprecipitation and gelatin zymography analyses, respectively. Wnt5a and LPA mediated MMP-9 regulation was assessed in OCSCC-derived SCC-9 cells exogenously expressing GSK3β (WT) or non phosphoryable GSK3β (S9A).

Results

We observed a progressive up-regulation/activation of MMP-9 at various stages of oral tumor progression/invasion. Positive correlations were observed between MMP-9 and c-Myc expression, MMP-9 and MMP-2 activity, MMP-9 and TIMP1 expression and MMP-9 activity and TIMP1-MMP-9 interaction. In contrast, a negative correlation between phosphorylated β-catenin and MMP-9 expression was observed. Conversely, we found that in oral tongue SCC MMP-9 expression was positively correlated with inactivation of GSK3 signaling. Finally, we found that Wnt5a and LPA mediated increased MMP-9 and decreased GSK3β activities in tongue SCC-derived SCC-9 cells. MMP-9 regulation by GSK3β was confirmed by using phosphoryable/regulatory GSK3β (WT construct) and not by non-phosphoryable GSK3β (S9A construct).

Conclusions

Collectively, our results show that MMP-9 overexpression and activation are important events occurring during OSCC progression/invasion and that this overexpression/activation is regulated by c-Myc, active MMP-2 and inactive GSK3β mediated pathways.

     

Garlic induces apoptosis during 7,12-dimethylbenz[a]anthracene-induced hamster buccal pouch carcinogenesis

The apoptosis-inducing capacity of aqueous garlic extract during 7,12-dimethylbenz[a]anthracene-induced hamster buccal pouch (HBP) carcinogenesis was investigated in male Syrian hamsters using DNA fragmentation and the apoptosis-associated proteins, tissue transglutaminase (tTG) and Bcl-2. Hamsters were divided into four groups of six animals each. Animals in group 1 were painted with a 0.5% solution of DMBA in liquid paraffin on the right buccal pouches three times a week for 14 weeks. Group 2 animals painted with DMBA as in group 1, in addition received 250 mg/kg body weight aqueous garlic extract orally on days alternate to DMBA application. Group 3 animals received garlic extract as in group 2. Group 4 animals received neither DMBA nor garlic extract and served as the control. The experiment was terminated at the end of 14 weeks. Administration of aqueous garlic extract (250 mg/kg body weight) to animals painted with DMBA inhibited DMBA-induced oral carcinogenesis as revealed by the absence of neoplasms, induction of tTG and inhibition of Bcl-2 expression. The results of the present study suggest that garlic may exert its chemopreventive effect by inducing apoptosis.     

Chlorophyllin abrogates canonical Wnt/β-catenin signaling and angiogenesis to inhibit the development of DMBA-induced hamster cheek pouch carcinomas

Background

Chlorophyllin, a water soluble semi-synthetic food-grade derivative is reported to exhibit a wide range of beneficial health effects. We investigated the effect of chlorophyllin supplementation on Wnt/??-catenin and vascular endothelial growth factor (VEGF) signaling in the 7,12-dimethylbenz[a]anthracene (DMBA)-induced hamster buccal pouch (HBP) carcinogenesis model.

Methods and results

Hamsters were divided into 4 groups. The right buccal pouches of group 1 and 2 hamsters were painted with 0.5?% DMBA for 14?weeks. Group 2 animals received in addition chlorophyllin (4?mg/kg bw) in the diet. Group 3 animals received chlorophyllin alone and group 4 animals served as control. mRNA and protein expression of components of Wnt, VEGF, and PI3K/Akt signaling pathways were analyzed by RT-PCR and Western blot analysis. Dietary chlorophyllin administration suppressed the development of HBP carcinomas by altering the expression of several components of the Wnt/??-catenin signaling pathway. This was associated with inhibition of angiogenesis as evidenced by decreased expression of the proangiogenic factors HIF-1??, VEGF, and VEGFR2. Chlorophyllin administration also downregulated the expression of histone deacetylases involved in epigenetic regulation of tumor angiogenesis.

Conclusion

Dietary chlorophyllin that abrogates Wnt/??-catenin and VEGF signaling by targeting a multitude of key signaling molecules is an attractive candidate for preventing tumor progression.