Tubeimoside-1 upregulates p21 expression and induces apoptosis and G2/M phase cell cycle arrest in human bladder cancer T24 cells

Abstract

Tubeimoside-1 (TBMS1) is a triterpenoid saponin with potent anticancer properties. In this study, for the first, we examined the anti-proliferative effects of TBMS1 in human bladder cancer T24 cells and its ability to induce apoptosis and cell cycle arrest. Our results demonstrated that TBMS1 decreased the cell viability of bladder cancer T24 cells in a dose-dependent manner. Flow cytometric analysis showed that TBMS1 significantly triggered apoptosis in T24 cells and arrested cell cycle at G2/M phase in a dosedependent manner. Further characterization demonstrated that TBMS1-induced apoptosis is associated with dissipation in mitochondrial membrane potential (ΔΨm), down-regulation of Bcl-2, and up-regulation of Bax and p21 in TBMS1-treated T24 cells. These in vitro results suggested that TBMS1 is an effective anti-bladder cancer natural compound that worth further mechanistic and therapeutic studies in human bladder cancer. Article InfoReceived:12 which is regulated by down-regulation of Bcl-2 and upregulation of Bax and p21. This study will pave the way for further in vivo studies of this potent anti-bladder cancer compounds. Material and Methods Chemicals and reagents:Cell culture medium reagents and MTT [3′-(4, 5 dimethyl-thiazol-2-yl)-2, 5-diphenyl tetrazolium bromide], propidium iodide (PI), and dimethyl sulfoxide (DMSO) were purchased from Sigma. Fetal bovine serum (FBS) was purchased from the Hangzhou Sijiqing Biological Engineering Materials Co., Ltd. Annexin V-FITC apoptosis detection kit was purchased from Beyotime Institute of Biotechnology Shanghai, China. Rabbit polyclonal anti-human Bcl-2, Bax, and p21 antibodies were purchased from Wuhan Boster Biological Technology Co., Ltd. β-actin, antimouse and anti-rabbit antibodies were purchased from Santa Cruz Biotechnology. Ponceou and cell lysis buffer for Western and IP were purchased from Bio SS Beijing. Rhodamine 123 was purchased from Eugene Co. (Oregon, U.S.A.).Cell culture: Human bladder cancer cell line (T24) was purchased from the Cell Bank of Shanghai Institute of Cell Biology, Chinese Academy of Sciences (Shanghai, China). The cells were cultured in DMEM supplemented with 10% FBS and 100 unit of Penicillin at 37℃ in a CO2 incubator with 5% CO2, 95% air and 100% humidity. Cells were plated in 10 cm culture dish and allowed to grow to approximately 70% confluence before experimentation.

Tubeimoside-1, Triterpenoid Saponin, as a Potential Natural Cancer Killer

“…JEG-3, T24, and A375 cells displayed significant G2/M phase arrest followed by the treatment Tubeimoside-1 as a potential natural cancer killer Natural Product Communications Vol. 13 (5) 2018 645 of tubeimoside-1 [40][41][42]. An investigation on anti-cancer activity against SKOV-3 cells has proved that pro-apoptotic property of tubeimoside-1 was accredited to G2/M phase arrest [43].…”

Anti-cancer effect of naringenin chalcone is mediated via the induction of autophagy, apoptosis and activation of PI3K/Akt signalling pathway

“…Annexin V-FITC assay (annexin V-FITC apoptosis detection kit) was used to quantify the extent of apoptosis induced by naringenin chalcone in U87MG human glioblastoma cells (Rasul et al, 2014). In brief, U87MG cells at a density of 2 x 10 5 cells/mL were seeded and then subjected to the treatment of naringenin chalcone at increasing doses (0, 10, 75 and 100 µM).…”