Induction of G2/M Cell Cycle Arrest and Apoptosis by Genistein in Human Bladder Cancer T24 Cells through Inhibition of the ROS-Dependent PI3k/Akt Signal Transduction Pathway

Abstract

We examined the anti-cancer effect of genistein, a soy-derived isoflavone, in human bladder transitional cell carcinoma T24 cells. According to our data, genistein induced G2/M phase arrest of the cell cycle and apoptosis. Genistein down-regulated the levels of cyclin A and cyclin B1, but up-regulated the levels of p21WAF1/CIP1, cyclin-dependent kinase (Cdk) inhibitor, that was complexed with Cdc2 and Cdk2. Furthermore, genistein induced the activation of caspases (caspase-3, -8 and -9), and cleavage of poly (ADP-ribose) polymerase cleavage. However, genistein-induced apoptosis was significantly inhibited by a pan-caspase inhibitor, indicating that the induction of apoptosis by genestein was caspase-dependent. In addition, genistein increased the cytosolic release of cytochrome c by increasing the Bax/Bcl-2 ratio and destroying mitochondria integrity. Moreover, genistein inactivated the phosphoinositide 3-kinase (PI3K)/Akt signaling pathway, while LY294002, a PI3K/Akt inhibitor, increased the apoptosis-inducing effect of genistein. Genistein further increased the accumulation of reactive oxygen species (ROS), which was significantly suppressed by N-acetyl cysteine (NAC), a ROS scavenger, and in particular, NAC prevented genistein-mediated inactivation of PI3K/Akt signaling, G2/M arrest and apoptosis. Therefore, the present results indicated that genistein promoted apoptosis induction in human bladder cancer T24 cells, which was associated with G2/M phase cell cycle arrest via regulation of ROS-dependent PI3K/Akt signaling pathway.

Trifolium pratense L. (red clover) extract and doxorubicin synergistically inhibits proliferation of 4T1 breast cancer in tumor‐bearing BALB/c mice through modulation of apoptosis and increase antioxidant and anti‐inflammatory related pathways

“…In a previous unpublished study on the compounds and minerals of red clover by LC‐ESI/MS and ICP‐ESI/SEM‐EDS, we found the main constituents of the plant as genistein, biohanin A, formononetin, quercetin, daidzein, luteolin, kaempferol, apigenin, epigallocatechin 3‐gallate polyphenols (flavonoids and isoflavonoids), and elements such as zinc, cobalt, selenium, manganese, and iron (Akbari Bazm et al, 2020). The flavonoids and isoflavones of this plant promote their anticancer effects via arresting the cell cycle at G1 or G2/M phases, inducing apoptosis, inhibiting ROS formation, metabolizing enzymes, and suppressing mediators such as vascular endothelial and basic fibroblast growth factors (Park et al., 2019). In our study, red clover extract, both alone and in synergy with DOX, increased the ratios of p53, Bax/Bcl‐2, and caspase‐3 in dose‐ and time‐dependent manners.…”

Pegylated recombinant human arginase 1 induces autophagy and apoptosis via the ROS-activated AKT/mTOR pathway in bladder cancer cells

“…The imbalance of ROS production and metabolism leads to oxidative stress, which is responsible for several pathological processes, such as autophagy, apoptosis, in ammation, DNA damage, aging and neurological diseases (28). Numerous chemotherapeutic drugs exert anti-tumor effects via ROSdependent cytotoxicity in cancer cells in vitro and in vivo (29)(30)(31). In our study, ROS production was boosted and glutathione (GSH) was correspondingly decreased in a dose dependent fashion after BCT-100 stimulation.…”

A Novel Peptide Oligomer of Bacitracin Induces M1 Macrophage Polarization by Facilitating Ca2+ Influx

“…RAW 264.7 cells were seeded on 6-well plates at a density of 4 × 10 5 cells/well, and incubated for 24 h. The cells were treated with CSP32 and l ng/mL LPS for 24 h, and then, the total RNA was isolated from the cells using TRIzol reagent (Invitrogen) according to the manufacturer’s instructions. cDNA was synthesized using the AccuPower ® PCR PreMix (Bioneer, Daejeon, Korea), as previously described [ 22 ]. The primer sequences used were as follows: cyclooxygenase-2 (COX-2; GenBank ID: NM-011198), sense (5′- GCGACATACTCAAGCAGGAGCA-3′) and antisense (5′- AGTGGTAACCGCTCAGGTGTTG-3′); glyceraldehyde 3-phosphate dehydrogenase (GAPDH, GenBank ID: NM-008084), sense (5′-CATCACTGCCACCCAGAAGACTG-3′) and antisense (5′-ATGCCAGTGAGCTTCCCGTTCAG-3′); IL-1β (GenBank ID: NM-008361), sense (5′-TGGACCTTCCAGGATGAGGACA-3′) and antisense (5′-GTTCATCTCGGAGCCTGTAGTG-3′); IL-6 (GenBank ID: NM-031168), sense (5′-TACCACTTCACAAGTCGGAGGC-3′) and antisense (5′- CTGCAAGTGCATCATCGTTGTTC-3′); inducible nitric oxide synthase (iNOS, GenBank ID: NM-010927), sense (5′- GAGACAGGGAAGTCTGAAGCAC-3′), and antisense (5′- CCAGCAGTAGTTGCTCCTCTTC-3′); and TNF-α (GenBank ID: NM-013693), sense (5′-GGTGCCTATGTCTCAGCCTCTT-3′) and antisense (5′-GCCATAGAACTGATGAGAGGGAG-3′).…”