Genistein induces apoptosis and autophagy in human breast MCF-7 cells by modulating the expression of proapoptotic factors and oxidative stress enzymes

Abstract

Breast cancer is one of the common tumors occurring in woman and despite treatment, the prognostic is poor. Genistein, a soy isoflavone, has been reported to have chemopreventive\chemotherapeutic potential in multiple tumor types. Here, we investigated the genistein antiproliferative effect in MCF-7 breast cancer, underlying the molecular mechanisms involved in this effect. MCF-7 cancer and CCD1059sK fibroblast cells were treated with estradiol (10 nM) or genistein (0.01-100 μM) for 24, 48, and 72 h and the cell proliferation was investigated by MTT; membrane cell permeability was evaluated by LDH and PI incorporation; apoptosis was investigated by externalization of phosphatidylserine by FACS; and presence of autophagy was detected by LC3A/B immunostaining. The expression of apoptotic proteins and antioxidant enzymes was evaluated by qPCR. The results demonstrate that genistein (100 μM) for 72 h of treatment selectively reduced MCF-7 cell proliferation independent of estrogen receptor activation, while no cytotoxicity was observed in fibroblast cells. Further experiments showed that genistein induced phosphatidylserine externalization and LC3A/B immunopositivity in MCF-7 cells, indicating apoptosis and autophagy cell death. Genistein increased in three times proapoptotic BAX/Bcl-2 ratio and promoted a parallel downregulation of 20 times of antiapoptotic survivin. In addition, genistein promoted a decrease of 5.5, 9.3, and 3.6 times of MnSOD, CuZnSOD, and TrxR mRNA expression, respectively, while the GPx expression was increased by 6.5 times. These results suggest that the antitumor effect of genistein involved the modulation of antioxidant enzyme and apoptotic signaling expression, which resulted in apoptosis and progression of autophagy.

Calycosin and Genistein Induce Apoptosis by Inactivation of HOTAIR/p-Akt Signaling Pathway in Human Breast Cancer MCF-7 Cells

“…In normal cells, Akt acts as a signal transducer of PI3K and promotes cell proliferation and cell survival. Activation of Akt favors cell survival through the regulation of apoptotic proteins, like the Bcl-2 family and caspases [20,21]. Thus, we believe that the observed downregulation of Akt phosphorylation may explain the mechanism for calycosin- and genistein-induced apoptosis.…”

Understanding genistein in cancer: The “good” and the “bad” effects: A review

“…In addition to genetic mutations, other important risk factors for the onset of BC are ''epigenetic variables", including lifestyle and nutrition. Since genistein modulates the estrogen-regulated gene expression and exerts a range of biological effects (Prietsch et al, 2014), it has become one of the most studied natural compounds to be tested in view of potential clinical applications against BC.…”

Apoptosis induction of colorectal cancer cells HTL-9 in vitro by the transformed products of soybean isoflavones by Ganoderma lucidum

“…Moreover, they can prevent and treat cardiovascular disease, osteoporosis (Srivastava et al, 2014;Baglia et al, 2015), breast cancer, prostate cancer (Li et al, 2012a), colon cancer (Tse and Eslick, 2016), and menopausal syndrome (Ollberding et al, 2012). The possible mechanisms of antitumor activity include an estrogenlike effect (Mense et al, 2008;Choi and Kim, 2013), the impact on the androgen receptor pathway by inhibiting the expression of prostate specific antigen (PSA) (Banerjee et al, 2012), the regulation of cell cycle and apoptosis induction (Prietsch et al, 2014;Tsuboy et al, 2014), and the regulation of mitogen activated protein kinase (MAPK) pathway (Cotrim et al, 2013;Chen et al, 2014).…”