Fig Effects of dietary intervention with sprouted chickpea diets

Fig. 2. Effects of dietary intervention with sprouted chickpea diets containing different amounts of selenium and isoflavonoids on tumor growth. (A) Relative areas of tumor growth starting day 9 after inoculation of HT-29 RFP cells. (B) In vivo imaging of tumor growth of HT-29 RFP obtained at day 15 post-inoculation. (C) Average tumor weight on day 21 after HT-29 RFP Lycopene xenografting. Control +: Positive control; Control −: Negative control; GC: Germinated control; GSe1: High content of selenium and isoflavonoids; GSe2: High content of selenium without presence of isoflavonoids. The values are means ± SE. Different letters above the bars denote significant differences (p < 0.05).
Fig. 3. Effects of dietary intervention with sprouted chickpea diets containing different amounts of selenium and isoflavonoids. (A) GPx activity. (B) TrxR activity. (C) Liver weight. At day 21 after inoculation with HT-29 RFP cells. GC: Germinated control; GSe1: High content of selenium and isoflavonoids; GSe2: High content of selenium without presence of isoflavonoids. Values are means ± SE. Different letters above the bars indicate significant differences (p < 0.05).
D. Guardado-Félix et al.
Fig. 4. Effects of dietary intervention with sprouted chickpea diets containing different amounts of selenium and iso-flavonoids on serum lipid levels (mg/dL) and expression of lipogenic genes at day 21 after inoculation with HT-29 RFP cells.
(A) CHOL: Cholesterol. (B) TRIGL: Triglycerides. (C) HDL-C: High-density lipoprotein cholesterol. (D) LDL-C: Low-density lipoprotein cholesterol. GC: Germinated control; GSe1: High content of selenium and isoflavonoids; GSe2: High content of selenium without pre-sence of isoflavonoids. The values are means ± SE. Different letters above the
bars indicate significant differences (p < 0.05).
(Fig. 4A–D). This was an indicator of the energy requirements necessary for cancer cell growth and proliferation. Mice fed with GSe1 and GSe2 diets had higher levels of CHOL, TRIGL and LDL-C compared to those fed GC diet (Fig. 4A–C) as an indication of the reversion of cancer cell proliferation. The groups fed with GC and GSe1 diets, rich in iso-flavonoids, showed HDL-C plasma levels higher than a diet without isoflavonoids (GSe2) (Fig. 4D). Therefore isoflavonoids in chickpea recovered the lipid metabolism in mice with cancer as it was previously reported for other polyphenolic compositions (Nandhakumar, Purushothaman, & Sachdanandam, 2014). No significant differences were observed between those treatments, indicating that there is no additive or synergistic effect due to the high selenium content of GSe1.
3.5. Effect of selenium and isoflavonoids on apoptotic gene and protein expression
The expression levels of 92 genes of Human p53 signaling were analyzed in tumor tissue and displayed on a heat map (Fig. 5A). The most significant transcriptional changes were observed in four genes: Caspase-9 (CASP9), hypoxia inducible factor 1 alpha subunit (HIF1A), apoptosis regulator (BCL2) and Fas cell surface death receptor (FAS) (Fig. 5A). CASP9, BCL2, FAS and HIF1A genes were suppressed from 2 to 5.7-fold in tumors of mice fed with experimental diets GSe1 and GSe2 compared to mice fed with GC diet (Fig. 5B).
At the translational level, the expression of Bcl2 in tumors from mice fed GSe1 diet was higher than in those fed with diet GC and GSe2 diets (Fig. 5C). Fas protein expression was higher in mice fed GSe1 diet even if the mRNA expression was lower compared to GC and GSe2 diets (Fig. 5C). An isoform of HIF1a was observed in the group fed with GSe1 diet (Fig. 5C). The expression level of caspase-9 protein decreased with GSe2 diet compared to GC and GSe1 diets (Fig. 5C).