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. 2016 Summer;5(3):178-191.
Epub 2016 Aug 30.

Genistein Induces Apoptosis and Inhibits Proliferation of HT29 Colon Cancer Cells

Gholamreza Shafiee  1 , Massoud Saidijam  2 , Heidar Tavilani  1 , Neda Ghasemkhani  1 , Iraj Khodadadi  1
Affiliations

Genistein Induces Apoptosis and Inhibits Proliferation of HT29 Colon Cancer Cells

Gholamreza Shafiee (VSports最新版本) et al. Int J Mol Cell Med. 2016 Summer.

Abstract

Soybean isoflavone genistein has multiple anticancer properties and its pro-apoptotic and anti-proliferative effects have been studied in different cancer cells. However, the mechanisms of action of genistein and its molecular targets on human colon cells have not been fully elucidated. Therefore, caspase-3 and p38 mitogen-activated protein kinase (p38 MAPK) as the main therapeutic targets were investigated in this study at both gene expression and protein levels in HT29 colon cancer cells. The caspase-3 and p38 MAPK gene expression levels were examined by real time PCR whereas flow cytometry technique was performed to determine their intracellular protein levels. The caspase-3 enzyme activity was obtained by colorimetric method while the gelatinase activity of matrix metalloproteinase-2 (MMP2) was determined by zymography. In addition, MTT test, wound healing assay and clonogenic assay were carried out to determine the effect of genistein on HT29 cell viability, migration, and proliferation, respectively. Genistein induced apoptotic death in HT29 cells through activation of caspase-3 pathway at the transcriptional, protein, and enzymatic levels. Moreover, genistein inhibited the proliferation of HT29 cells by reducing of both p38 MAPK gene expression and its active phosphorylated protein level VSports手机版. Also, we showed that genistein strongly suppressed the metastatic potency of HT29 colon cancer cells via the reduction of MMP2 activity. Based on the results of this study, we conclude that genistein may exhibit its anticancer properties on HT29 colon cancer cells by modulating caspase-3 and p38 MAPK pathway at different transcriptional and protein levels. .

Keywords: Apoptosis; caspase-3; colonic neoplasms; genistein; p38 mitogen-activated protein kinase. V体育安卓版.

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Figures

Fig. 1
Fig. 1
The effect of genistein on HT29 cell viability. Exposure of HT29 cells to different concentration of genistein for 12, 24, and 48 h decreased cell viability in a dose-dependent manner. The incubation of the cells with genistein for 48 h exhibited greater decline in the cell viability compared with shorter incubation time
Fig. 2
Fig. 2
The effects of genistein on HT29 cell migration analyzed by wound healing assay. Cells were cultured with 0, 10, 30 and 50 µM genistein for 0, 12, 24 and 48 h, a wound track was scored and the number of migrated cells was determined. Top: the number of migrated cells was significantly reduced after exposure of HT29 cells to 50 µM genistein for 48 h (e-h) compared with untreated control group (a-d). Bottom: wound diameter (µm) was determined and One-Way-ANOVA analysis was performed to compare differences between groups. The symbol (*) represents significant differences (P< 0.05) between groups compared to untreated cells
Fig. 3
Fig. 3
The effects of genistein on the proliferation of HT29 cells analyzed by colony formation assay. Cells were plated and treated with 0, 10, 30 and 50 µM genistein for one week. The number of colonies was determined, plating efficiency was calculated and the percentage of survival factor was obtained. Figure shows the stained cultured cells (top) and survival factor (%) based on different concentration of genistein (bottom). One-Way-ANOVA analysis was performed to compare the differences between groups and the symbol (*) represents significant differences (P< 0.05) between groups compared to untreated cells
Fig. 4
Fig. 4
Dose-dependent proapoptotic effects of genistein on HT29 cells, as determined by RT-PCR and flow cytometry. The relative gene expression of caspase-3 in HT29 cells upon exposure to 0, 30, 50 and 70 µM genistein was examined by RT-PCR. Caspase-3 gene expression was increased by increasing genistein concentrations compared with 18S rRNA gene expression (top). Genistein also induced significant apoptosis in HT29 cells by increasing caspase-3 protein compared to untreated cells, as determined by flow cytometry (bottom). One-Way-ANOVA analysis was performed to compare differences between groups. (*) and (**) indicate significant difference at P< 0.05 and P< 0.01 levels compared with untreated cells, respectively
Fig. 5
Fig. 5
Dose-dependent antiproliferative effects of genistein on HT29 cells, as determined by RT-PCR and flow cytometry. The relative gene expression of p38 MAPK in HT29 cells upon exposure to 0, 30, 50 and 70 µM genistein was examined by RT-PCR. P38 MAPK gene expression was reduced by increasing genistein concentrations compared with 18S rRNA gene expression (top). Genistein also significantly inhibited HT29 cells proliferation by reducing p-p38 MAPK protein compared to untreated cells, as determined by flow cytometry (bottom). One-Way-ANOVA analysis was performed to compare differences between groups. (*) and (**) indicate significant difference at P< 0.05 and P< 0.01 levels compared with untreated cells, respectively
Fig. 6
Fig. 6
The effect of 0, 30, 50 and 70 µM genistein on MMP2 caspase-3 enzyme activities in HT29 cells. Top: cells were incubated for 48 h and conditioned media were used for the measurement of MMP2 activity by gelatin zymography. The enzyme activity (MMP2) was reduced by increasing genistein concentrations. Bottom: caspase-3 activity was significantly increased with increasing genistein concentrations and the greatest enhancement in caspase-3 activity (89%) was observed in the presence of 70 µM genistein. One-Way-ANOVA analysis was performed to compare differences between groups and the symbol (*) represents significant differences (P< 0.05) between groups compared to untreated cells
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References

    1. Arber N, Levin B. Chemoprevention of colorectal cancer: ready for routine use? Recent results in cancer research Fortschritte der Krebsforschung Progres dans les recherches sur le cancer. 2005;166:213–30. - PubMed
    1. Bray F, Ferlay J, Laversanne M, et al. Cancer Incidence in Five Continents: Inclusion criteria, highlights from Volume X and the global status of cancer registration. International journal of cancer. 2015;137:2060–71. - PubMed
    1. Thompson R. Preventing cancer: the role of food, nutrition and physical activity. The journal of family health care. 2010;20:100–2. - PubMed
    1. Song WO, Chun OK, Hwang I, et al. Soy isoflavones as safe functional ingredients. Journal of medicinal food. 2007;10:571–80. - PubMed
    1. Azeem S, Gillani SW, Siddiqui A, et al. Diet and Colorectal Cancer Risk in Asia--a Systematic Review. Asian Pacific journal of cancer prevention : APJCP. 2015;16:5389–96. - PubMed