Contribution of p53-mediated Bax transactivation in theaflavin-induced mammary epithelial carcinoma cell apoptosis

Abstract

Theaflavins, the bioactive flavonoids of black tea, have been demonstrated to inhibit proliferation and induce apoptosis in a variety of cancer cells. However, the contribution of p53 in mammary epithelial carcinoma cell apoptosis by theaflavins remains unclear. It has been reported that p53 triggers apoptosis by inducing mitochondrial outer membrane permeabilization through transcription-dependent and -independent mechanisms. Using wild-type and mutant p53-expressing as well as p53-null cells we found a strong correlation between p53 status and theaflavin-induced breast cancer cell apoptosis. Apoptogenic effect was more pronounced in functional p53-expressing cells in which theaflavins raised p53 protein levels that harmonized with Bax up-regulation and migration to mitochondria. However, in the same cells, when p53-mediated transactivation was inhibited by pifithrin-alpha, theaflavins not only failed to increase transcription but also to induce apoptosis although p53 up-regulation was not altered. In contrast, Bax over-expression restored back theaflavin-induced apoptosis in pifithrin-alpha-inhibited/dominant-negative p53-expressing cells. Inhibition of Bax by RNA-interference also reduced theaflavin-induced apoptosis. These results not only indicated the requirement of p53-mediated transcriptional activation of Bax but also its role as down-stream effecter in theaflavin-induced apoptosis. Bax up-regulation resulted in mitochondrial transmembrane potential loss and cytochrome c release followed by activation of caspase cascade. In contrast, mitochondrial translocation of p53 and its interaction with Bcl-2 family proteins or activation of caspase-8 could not be traced thereby excluding the involvement of p53-mediated transcription-independent pathways. Together these findings suggest that in breast cancer cells, p53 promotes theaflavin-induced apoptosis in a transcription-dependent manner through mitochondrial death cascade.

Curcumin Enhances the Efficacy of Chemotherapy by Tailoring p65NFκB-p300 Cross-talk in Favor of p53-p300 in Breast Cancer

“…Reverse Transcriptase and Real-time Quantitative RT-PCR Analysis-Two g of the total RNA each from doxorubicin/ curcumin-treated doxorubicin-sensitive/-resistant cells was extracted by TRIzol (Invitrogen) and reverse-transcribed and then subjected to PCR with enzymes and reagents of the RTplusPCR system (Eppendorf, Hamburg, Germany) using GeneAmpPCR 2720 (Applied Biosystems Foster City, CA) (39,40). The cDNAs were amplified with primers specific for Bax (5Ј-GGAATTCCAAGAAGCTGAGCGAGTGT-3Ј/5Ј-GGA-ATTCTTCTTCCAGATGGTGAGCGAG-3Ј), PUMA (5Ј-AGCTCCCATCCTGGCTCTGG-3Ј/5Ј-CAGGCAGTTGT-CAGCTGGG-3Ј, Noxa (5Ј-ACTGTGGTTCTGGCGCAGAT-3Ј/5Ј-TGAGCACACTCGTCCTTCAAGT-3Ј), Bcl-2 (5Ј-CCTGTGCCACCATGTGTCCATC-3Ј/5Ј-GCTGAGAAC-AGGGTCTTCAGAGAC-3), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH; internal control, 5Ј-TGATGACAT-CAAGAAGGTGGTGAAG-3Ј/5Ј-TCCTTGGAGGCCATGT-AGGCCAT-3).…”

Cyanide-Induced Apoptosis of Dopaminergic Cells Is Promoted by BNIP3 and Bax Modulation of Endoplasmic Reticulum-Mitochondrial Ca2+ Levels

“…jpet.aspetjournals.org remain unchanged after cyanide treatment . Under normal conditions Bax is a cytosolic protein that translocates to the ER/mitochondria in response to apoptotic stimuli (Lahiry et al, 2008;Liao et al, 2008). On the other hand, overexpressed BNIP3 can localize to both ER and mitochondria (Ray et al, 2000;Zhang et al, 2009).…”

Effect of Black Tea Polyphenol on Cell-ECM Interaction and MMP

“…Treatment with black tea polyphenols has been shown to induce apoptosis in prostate cancer cells by induction of p53 and down-regulation of NF-κB activity [36]. Theaflavins have been reported to induce apoptosis in mammary epithelial carcinomas through inhibition of PI-3K-Akt-pBad survival pathway [37]. Theaflavin and extracts from black tea have been shown to induce apopotosis in human leukemic cell lines HL-60 and K-562 [40].…”