Cell Cycle Modulation of CHO-K1 Cells Under Genistein Treatment Correlates with Cells Senescence, Apoptosis and ROS Level but in a Dose-Dependent Manner

Abstract

Purpose: Genistein, a soy isoflavone, exhibits a biphasic effect on cells proliferation with some different effects between ER-alpha and ER-beta. The objective of this present study is to determine the modulatory effect based on cell cycle progression under genistein treatment in combination with 17-β estradiol (E2) on CHO-K1 cells. Methods: The effect of genistein 0.1-100 µM on cells proliferation was examined by MTT assay. The modulation of genistein and estradiol (E2) on cell cycle and apoptosis were observed by using flowcytometry with PI and PI/AnnexinV staining, respectively. Moreover, the effect of genistein and E2 on senescence cells, and ROS level were determined by senescence-associated β-galactosidase (SA β-gal) staining and by using flowcytometry with 2’, 7’–dichlorofluorescin diacetate (DCFDA) staining, respectively. The expression level of the cell cycle and senescence protein markers were observed by immunoblotting. Results: Single treatment of genistein at physiologically achievable (low) concentration (<2 µM) induced proliferation of CHO-K1 cells while at a pharmacological (high) concentration (50 and 100 µM) suppressed cells proliferation. Interestingly, treatment of genistein at the physiological concentration in combination with E2 for 24, 48 and 72 h decreased cells viability on CHO-K1 cells compared to untreated cells. Further analysis of the cells showed that 50 µM genistein induced G2/M phase accumulation and induced apoptosis. Moreover, genistein induced cell senescence and increased ROS level. Immunoblotting analysis showed the decreasing of ERalpha, Bcl2, and ppRb protein level upon treatment of 1 µM Gen and 1 nM E2. Conclusion: Our results suggest that the cell proliferation inhibitory mechanism of genistein at pharmacological concentration involved the induction of cell senescence, and the elevation of ROS level. Moreover, the decreased of cells proliferation upon treatment of physiological concentration of genistein in combination with E2 may be correlated with the alteration of ER expression.

Citrus sinensis (L) Peels Extract Inhibits Metastasis of Breast Cancer Cells by Targeting the Downregulation Matrix Metalloproteinases-9

“…A single treatment of CSP and DOX individually possessed strong cytotoxic effect with dosedependent manner and the low dose of DOX induced lamellipodia elongation. This phenomenon supports the previous studies that DOX in concentration of 10 nM induced lamellipodia formation through the activate transforming growth factor beta (TGFβ) signaling pathway [9], [22]. The activation of TGFβ induces EMT caused inducing motility of cell and metastatic [8].…”

Synergistic Effect of Curcuma longa Extract in Combination with Phyllanthus niruri Extract in Regulating Annexin A2, Epidermal Growth Factor Receptor, Matrix Metalloproteinases, and Pyruvate Kinase M1/2 Signaling Pathway on Breast Cancer Stem Cell

“…where D 1 and [Dx] 1 are concentrations of CL Rhizome and PN herbs, respectively, which inhibit cell growth to 50% of control when used alone, and D 2 and [Dx] 2 are concentrations of CL rhizome and PN herbs, respectively, which have the same effect when used in combination [33]. Combination index values based on the Chou-Talalay method, calculated using CompuSyn software, indicate the effects of drug combinations [34], [35].…”

Antiproliferative and apoptosis effect of hyptolide from Hyptis pectinata (L.) Poit on human breast cancer cells

“…Similar results were performed by previous studies in which reported (Tamura et al, 2009;Usui et al, 2004) that pironetin (have a similar structure with hyptolide) could inhibit the growth of HT1080 and 3Y1 cells by apoptosis induction. The apoptosis induction on ER-positive breast cancer cells is due to the role of ER in inhibiting cell growth (Jenie et al, 2019;Rouhimoghadam et al, 2018). According to this finding, cell viability response that under hyptolide treatment showed a dose-dependent phenomenon and most of the cell deaths occurred predominantly through apoptosis.…”

“…Blocking the action of estrogen has been shown to reduce breast cancer proliferation and incidence in high-risk women by 50%-75% (Lewis-Wambi and Jordan, 2009;Tian et al, 2018). The previous study also found that estrogen administration causes breast cancer while antiestrogen prevents it (Hollingsworth et al, 1998;Jenie et al, 2019). Those findings provide compelling evidence that estrogen and ER have a significant role in the progression of breast cancer.…”