AIM: To investigate the role of quercetin in the apoptosis of human breast cancer cell line MCF-7 and the association with Fas/Fas ligand (FasL) pathway. METHODS: The apoptosis model of MCF-7 cells was established by the induction with quercetin. The morphological characteristics of apoptotic MCF-7 cells were observed under transmission electron microscope. The apoptotic rates and alternation of mitochondrial membrane potential (Δψm) in the MCF-7 cells were measured by flow cytometry using fluorescein labeled Annexin V-FITC/PI and JC-1, respectively. FasL neutralizing antibody was applied to block the apoptosis. The expression of Fas/FasL on the cells was detected by immunofluorescence technique and flow cytometry, respectively. The influence of SB203580 (an inhibitor of p38 MAPK) on the expression of Fas/FasL was also examined by flow cytometry. The protein levels of p38 MAPK and p-p38 MAPK were determined by Western blot. RESULTS: The phenomenon of nuclear condensation and marginalization in the MCF-7 cells treated with quercetin at 80.0 μmol/L for 48 h was observed under transmission electron microscope. Compared with the control cells, the Δψm was decreased by 17.4%, 44.3% and 68.9% in the MCF-7 cells treated with quercetin at 80.0 μmol/L for 24 h, 48 h and 72 h, respectively. The apoptotic rates of MCF-7 cells treated with quercetin at 80.0 μmol/L for 24 h, 48 h and 72 h were (10.2±3.3)%, (28.9±7.5)% and (39.2±8.9)%, respectively. However, the apoptotic rates were decreased to (8.2±2.8)%, (19.2±5.3)% and (22.5±6.9)% after the cells were pretreated with FasL neutralizing antibody, respectively. When MCF-7 cells were treated with quercetin for 24 h, 48 h and 72 h, Fas/FasL expression rates were increased in a time-dependent manner, which were largely inhibited by SB203580. The protein level of p38 MAPK was not changed obviously, but the protein level of p-p38 MAPK was significantly increased at 48 h and 72 h. CONCLUSION: Quercetin up-regulates the expression of Fas/FasL on MCF-7 cells, and induces apoptosis via Fas/FasL pathway. Meanwhile, p-p38 MAPK is potentially critical signaling molecule for up-regulating the expression of Fas/FasL.
Chen WS, Wang X, Zhuang JG, et al. Induction of death receptor 5 and suppression of survivin contribute to sensitization of TRAIL-induced cytotoxicity by quercetin in non-small cell lung cancer cells[J]. Carcinogenesis, 2007, 28(10):2114-2121.
Mediero JM, Alonso A, Borda A, et al. Effect of polyphenols from the Mediterranan diet on proliferation and mediators of in vitro invasiveness of the MB-49 murine bladder cancer cell line[J]. Actas Urol Esp, 2005, 29(8): 743-749.
[6]
Zhou J, Liang S, Fang L, et al. Quantitative proteomic analysis of HepG2 cells treated with quercetin suggests IQGAP1 involved in quercetin-induced regulation of cell proliferation and migration[J]. OMICS, 2009, 13(2):93-103.
[7]
Duo J, Ying GG, Wang GW, et al. Quercetin inhibits human breast cancer cell proliferation and induces apoptosis via Bcl-2 and Bax regulation[J]. Mol Med Rep, 2012, 5(6):1453-1456.
Chou CC, Yang JS, Lu HF, et al. Quercetin-mediated cell cycle arrest and apoptosis involving activation of a caspase cascade through the mitochondrial pathway in human breast cancer MCF-7 cells[J]. Arch Pharm Res, 2010, 33(8):1181-1191.
[11]
Rudner J,Elsaesser SJ, Jendrossek V, et al. Anti-apopto-tic Bcl-2 fails to form efficient complexes with pro-apopto-tic Bak to protect from Celecoxib-induced apoptosis[J]. Biochem Pharmacol, 2011, 81(1):32-42.
[12]
Zhang D, Wang L, Yan L, et al. Vacuolar protein sorting 4B regulates apoptosis of intestinal epithelial cells via p38 MAPK in Crohn's disease[J]. Exp Mol Pathol, 2015, 98(1):55-64.
[13]
Wang M, Li Y, Wu M, et al. WWOX suppresses cell growth and induces cell apoptosis via inhibition of P38 nuclear translocation in cholangiocarcinoma[J]. Cell Physiol Biochem, 2014, 34(5):1711-1722.
[14]
Urriola-Muñoz P, Lagos-Cabré R, Morenio RD. A mechanism of male germ cell apoptosis induced by bisphenol-A and nonylphenol involving ADAM17 and p38 MAPK activation[J]. PLoS One, 2014, 9(12):e113793.
