Combined Effects Of Genistein Research Articles

Inhibitory effects of genistein in combination with gefitinib on the hepatocellular carcinoma Hep3B cell line.

Combination therapy is an important method for treating advanced hepatocellular carcinoma (HCC). Gefitinib is an epidermal growth factor receptor (EGFR) inhibitor, which has profound effects on HCC. The purpose of the present study was to investigate the effects of genistein in combination with gefitinib on the proliferation and apoptosis of HCC cells and the associated mechanism. Cell counting kit-8 assay was performed to calculate the IC50 values and cytotoxicity, whilst flow cytometry was used to assess cell apoptosis. Protein expression was detected using western blot analysis. The IC50 of genistein and gefitinib on Hep3B cells were calculated to be 128.078 and 13.657 µM, respectively. Genistein in combination with gefitinib significantly inhibited cell viability, promoted apoptosis and reduced EGFR, vascular endothelial growth factor receptor and platelet-derived growth factor receptor phosphorylation. Genistein in combination with gefitinib promoted the expression of cleaved caspase-3 and cleaved poly ADP-ribose polymerase. In addition, combined treatment of genistein and gefitinib strongly inhibited the activation of the Akt/Erk/mTOR signaling pathway. In conclusion, findings from the present study suggest that genistein in combination with gefitinib inhibit HCC cell proliferation and promote apoptosis by inhibiting the Akt/Erk/mTOR pathway.

RETRACTED ARTICLE: Genistein inhibits stemness of SKOV3 cells induced by macrophages co-cultured with ovarian cancer stem-like cells through IL-8/STAT3 axis

BackgroundRecent studies showed that macrophages co-cultured with ovarian cancer stem-like cells (OCSLCs) induced SKOV3 cell stemness via IL-8/STAT3 signaling. Genistein (GEN) demonstrates chemopreventive activity in inflammation-associated cancers. The present study aimed to examine whether and if GEN inhibits the stemness of SKOV3 and OVCA-3R cells induced by co-culture of THP-1 macrophages and SKOV3-derived OCSLCs.MethodsThe co-culture was treated with or without different concentrations (10, 20, and 40 μmol/L) of GEN for 24 h. Depletion or addition of IL-8 in Co-CM and knockdown or overexpression of STAT3 in THP-1 macrophages was performed to demonstrate the possible associated mechanisms. The combined effects of GEN and STAT3 knockdown were examined with the nude mouse modle by co-injection of SKOV3-derived OCSLCs with THP-1 macrophages.ResultsOur results showed that GEN down-regulated CD163 and p-STAT3 expression of THP-1 macrophage, decreased the levels of IL-10, increased the levels of IL-12 and nitric oxide (NO) in the conditioned medium, and reduced the clonogenic and sphere-forming capacities and the expression of CD133 and CD44 in SKOV3 cells induced by co-culture of THP-1 macrophages and OCSLCs in a dose-dependent manner. Moreover, depletion or addition of IL-8 enhanced or attenuated the effect of GEN. Additionally, knockdown or overepression of STAT3 in THP-1 macrophages potentiated or attenuated the inhibitory effects of GEN. Importantly, STAT3 overexpression retrieved the effects of IL-8 combined with GEN depletion on M2 polarization of THP-1 macrophages and stemness of SKOV3 cells induced by co-culture. The combination of GEN and STAT3 knockdown cooperatively inhibited the growth of tumors co-inoculated with OCSLCs/THP-1 macrophages in nude mice in vivo through blocking IL-8/STAT3 signaling.ConclusionsIn summary, our findings suggested that GEN can inhibit the increased M2 polarization of macrophages and stemness of ovarian cancer cells by co-culture of macrophages with OCSLCs through disrupting IL-8/STAT3 signaling axis. This assisted GEN to be as a potential chemotherapeutic agent in human ovarian cancer.

Abstract 909: Mechanistic study of synergistic interaction between genistein and equol in MCF-7 human breast cancer cells in vitro

Abstract Soy isoflavones, such as genistein, daidzein, glycitein and equol, a metabolite of daidzein, have been suggested to affect the growth of estrogen-receptor positive breast cancer cells in vitro, and thus considered to be involved in the prevention of breast cancer. We have recently reported that soy isoflavone component mixture has a greater preventive activity against breast cancer rather than genistein alone using chemically induced rat model of hormone-dependent mammary carcinoma. Therefore, we investigated here whether a synergistic interaction exists between genistein and other isoflavone components in estrogen receptor positive MCF-7 cells. Effects of genistein in combination with other isoflavone components on proliferation, apoptosis, cell cycle perturbation, and signal proteins for survival, proliferation, and apoptosis were evaluated by WST-1 assay, FACS analyses, and Western blotting. Synergistic anti-proliferative activity of genistein was observed in combination with three respective isoflavone components, with the most efficacious interaction observed in combination with equal. In FACS study, cells were arrested at the G2/M by genistein but G1/S by equol. However, when these components were combined, cells were arrested at the G2/M, indicating a dominant effect of genistein over equol. Although apoptosis was not evident when cells were treated with either genistein or equol alone, combination treatment markedly induced apoptosis, with demonstrable cleavage of PARP. Upon combination treatment, constitutive activities of Erk1/2 were enhanced, but those of Akt were significantly inhibited, with subsequent reductions of activities of downstream mTOR. Reduced expression of anti-apoptotic Bcl-xL and enhanced expression of proapoptotic Bax were also demonstrated. These data indicate that synergic mechanism of genistein in combination with equol would be mediated by enhanced apoptosis induction through inactivation of Akt and its downstream molecule mTOR, followed by subsequent upregulation of Bax and downregulation of Bcl-xL, thus providing mechanistic rationale for soy isoflavone for the prevention of human breast cancer. Citation Format: MISAKI ONO, KAORU EJIMA, MIKAKO TAKESHIMA, TAKAKO HIGUCHI, REI WAKIMOTO, SHUJI NAKANO. Mechanistic study of synergistic interaction between genistein and equol in MCF-7 human breast cancer cells in vitro. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 909. doi:10.1158/1538-7445.AM2015-909

Individual and combined effects of genistein and hesperidin on immunity and intestinal morphometry in lipopolysacharide-challenged broiler chickens

Genistein and hesperidin have been shown to have beneficial effects in several animal models including mice, rats, pigs, and humans. This study investigated the individual and combined effects of genistein (an isoflavone) and hesperidin (a flavanone) on immunity and intestinal morphometry in lipopolysaccharide (LPS)-challenged broiler chickens. Seven hundred twenty 1-d-old commercial Arbor Acres broiler chicks were randomly divided into 6 treatments, with 6 replicates of 20 birds each. Chicks were fed a basal diet without any additive (control), supplemented with 5 mg of genistein/kg of feed (G5) and 20 mg of hesperidin/kg of feed (H20), or a mixture of genistein and hesperidin (1:4) with doses of 5 (GH5), 10 (GH10), and 20 (GH20) mg/kg of feed for 42 d. On d 16, 18, and 20, one-half the birds from each group were separated and injected intraperitoneally with Escherichia coli LPS (250 µg/kg of BW) to induce the immunological stimulation. Samples were collected on 21 and 42 d. The results showed that LPS treatment exerts immunomodulatory effects (P < 0.05) in phagocytic activity at 21 d, whereas a few negative effects including reduced villus length and increased crypt depth were observed in some segments of the small intestine. Both genistein and hesperidin seemed to modify the immunity positively by altering the phagocytic activity (P < 0.01). Parameters of intestinal morphometry such as villus length, crypt depth, villus width, and villus length/crypt depth ratios were also improved (P < 0.01 or P < 0.05) by the supplemental genistein and hesperidin in both LPS-unchallenged and -challenged groups. However, no effect (P > 0.05) was observed for BWG, FI, and FCR of broilers. Overall, genistein and hesperidin improved the immunity and the morphometry of small intestine in a dose-dependent manner. These findings provided the first account on the in vivo effects of genistein and hesperidin for immunostimulation and morphometric gut development in LPS-challenged chickens. Thus, both compounds may be used as alternative feed additives in the poultry industry to promote gut health and improve immunity against infections.

Individual and combined effects of genistein and hesperidin supplementation on meat quality in meat‐type broiler chickens

There is growing interest in improving the production and meat quality of farm animals through dietary supplementation with phytochemical (e.g. flavonoids)-rich plants and/or their extracts. This study was conducted to analyse the supplemental effects of two purified flavonoids (genistein and hesperidin) individually and in combination on the oxidative status, sensory score and quality of breast meat in meat-type broiler chickens. A significant increase (P < 0.05) in meat colour (L* score) and pH was observed for the group supplemented with 20 mg kg(-1) genistein and hesperidin. Water-holding capacity was also improved significantly (P < 0.01) for all genistein- and hesperidin-treated groups, while the sensory quality of breast meat remained unaffected. Lipid oxidation of breast meat was reduced significantly (P < 0.01) at 0 and 15 days of refrigeration in a dose-dependent manner for all supplemented groups. Meanwhile, some treated groups showed improved (P < 0.05) body weight, feed/gain ratio and hot carcass weight. Genistein and hesperidin supplementation to broilers improved meat quality in a dose-dependent fashion, with pronounced effects of combined treatment. The results indicated that purified flavonoids such as genistein and hesperidin could potentially be used as feed additives in broiler production to promote meat quality.

Anti‐inflammatory and Anticarcinogenic Effect of Genistein Alone or in Combination with Capsaicin in TPA‐Treated Rat Mammary Glands or Mammary Cancer Cell Line

The topical application of TPA (12-O-tetradecanoylphorbol-13-acetate) to animal skin or direct treatment of TPA to cell cultures leads to inflammatory responses by enhancing cyclooxygenase 2 (COX-2) expression, and specific COX-2 inhibitors counteract this kind of inflammatory response. Furthermore, suppression of these inflammatory events by dietary-origin chemopreventive agents can provide a potential strategy to control carcinogenesis. In this in vivo study, the mammary glands of mature female rats were treated with TPA, and then the effects of genistein alone or in combination with capsaicin on suppression of inflammatory responses were examined. The combined effects of genistein and capsaicin on COX-2, pJNK, pERK, and pp38 expressions were additive or nonadditive, depending on signals tested. In vitro MCF-7 breast cancer cells, the apoptotic bodies as shown with Hoechst 33342 dye, exhibited a synergistic effect between genistein and capsaicin. The abilities of genistein alone or in combination with capsaicin in inhibiting breast cancer cell proliferation through the modulation of AMPK and COX-2 were tested. AMPK activation by genistein in combination with capsaicin is critical for inhibiting COX-2. We propose that genistein in combination with capsaicin exerts anti-inflammatory and anticarcinogenic properties through the modulation of AMPK and COX-2 and possibly various mitogen-activated protein kinases synergistically or nonsynergistically.

Genistein inhibits Vitamin D hydroxylases CYP24 and CYP27B1 expression in prostate cells

In human prostate cancer cells, the availability of the steroid hormone 1,25-dihydroxyvitamin D 3 for antimitotic action is determined through the activity of the two enzymes CYP24 and CYP27B1, viz. 25-hydroxyvitamin D-24-hydroxylase and 25-hydroxyvitamin D-1α-hydroxylase. High performance liquid chromatography (HPLC) analysis of [ 3H]25(OH)D 3 metabolism in human prostate cancer DU-145 cells revealed that genistein and other isoflavonoids, such as dihydrogenistein and daidzein, as well as the antiestrogenic compound ICI 182,780, inhibited Vitamin D-metabolizing enzyme activities. Reverse transcriptase-polymerase chain reaction (RT-PCR) showed that only in case of genistein this was due to transcriptional inhibition of CYP24 and CYP27B1 gene expressions. In case of CYP27B1, reduction of gene activity involves histone deacetylation because genistein was inactive in the presence of the histone deactylase inhibitor trichostatin A. In contrast, under the same condition, CYP24 gene activity was largely suppressed. In summary, our results suggest that a combined effect of genistein and trichostatin A could increase the responsiveness of human prostate cancer cells to the antiproliferative action of 1,25-dihydroxyvitamin D 3.