Annonacin induces cell cycle-dependent growth arrest and apoptosis in estrogen receptor-α-related pathways in MCF-7 cells
Graphical abstract
Introduction
The pathogenesis of breast cancer includes estrogen and estrogen receptor-α (ERα)-related pathways (Osborne and Schiff, 2011) whereby nuclear ERα activates target genes via the estrogen-response elements (ERE) (Osborne and Schiff, 2011). Additionally, nonnuclear ERα rapidly activates growth factor downstream signals such as extracellular signal-regulated kinases (ERK1/2), c-Jun N-terminal kinase (JNK) and signal transducers and activators of transcription 3 (STAT3) (Musgrove and Sutherland, 2009, Osborne and Schiff, 2011). Interestingly, STAT integrates cytoplasmic and nuclear estrogen actions (Fox et al., 2009).
Selective estrogen receptor modulators (SERM, such as tamoxifen) or selective estrogen receptor down-regulators (SERD, such as fulvestrant) are effective treatments for ERα-positive breast cancers commonly limited by resistance (Osborne and Schiff, 2011). Endocrine resistance may be induced by the loss of ERα, increased nonnuclear ERα or growth factor receptor signaling, deranged signal transducers (such as ERK1/ERK1, JNK and STAT3), cell cycle regulators (such as cyclin D1 and the cyclin-dependent kinase inhibitors p21WAF1 and p27kip1) or apoptosis regulators (such as the anti-apoptotic Bcl-2) (Musgrove and Sutherland, 2009, Osborne and Schiff, 2011).
Pawpaw (Asimina triloba) and soursop (graviola, Annona muricata) are annonaceous plants used as anticancer folk therapies in (North, Central and South) America and Southeast Asia and have been studied in a few observational clinical studies (Cassileth, 2008, Coothankandaswamy et al., 2010, Liaw et al., 2010, McLaughlin, 2008). Annonaceous acetogenins are cytotoxic to multidrug-resistant MCF-7 cells (Oberlies et al., 1997). Annonacin (C35H64O7), an annonaceous acetogenin containing a mono-tetrahydrofuran ring with two flanking hydroxyls, also inhibits growth in MCF-7 cells (Yuan et al., 2003). However, the molecular mechanisms are not understood.
Thus, we studied the growth-inhibitory mechanisms of annonacin in terms of ERα-related pathways (p-ERK1/2, p-JNK, p-STAT3, cyclin D1, Bcl-2, p21WAF1 and p27kip1) in MCF-7 cells. Moreover, the effects of annonacin on MCF-7 xenografts in nude mice were also investigated.
Section snippets
Cell culture and reagents
ERα-positive MCF-7 cells and ERα-negative MDA-MB-231 cells were purchased on February 18, 2009 from Bioresource Collection and Research Center (Hsinchu, Taiwan), where cells were authenticated by DNA fingerprints of short tandem repeat profiling. Cells were cultured in DMEM/F-12 (1:1) medium supplemented with 1% penicillin/streptomycin and 10% fetal bovine serum (FBS, Gibco, Grand Island, NY, USA) in a humidified 5% CO2 incubator at 37 °C. Cells were fasted for 24 h before adding fresh medium
Effects of annonacin on cell cycle distribution and cell death in MCF-7 cells
Annonacin (0.1 μM) time-dependently (6–48 h) arrested cells in the G0/G1 phase of the cell cycle (Fig. 1A) while increasing apoptosis at 48 h (Fig. 1B). Additionally, annonacin dose-dependently (0.5–1 μM) increased cell death at 48 h (Fig. 1C). Annonacin also dose-dependently (0.05–1 μM) decreased cell survival at 48 h (supplementary Fig. 2A). Moreover, annonacin (0.1 μM) time-dependently (24–48 h) decreased cell survival (supplementary Fig. 2B). However, annonacin (0.1 μM) did not affect cell survival
Discussion and conclusions
This study adds mechanistic insights to our previous finding that annonacin inhibits growth in MCF-7 cells (Yuan et al., 2003).We found that annonacin induced cell death only at high doses (≥0.5 μM). In contrast, low-dose (0.1 μM) annonacin, like tamoxifen and fulvestrant (Cariou et al., 2000, Musgrove and Sutherland, 2009), induced cell-cycle-dependent (G0/G1) growth arrest concomitantly with the induction of p21WAF1 and p27kip and the inhibition of cyclin D1 protein expression.
Cell cycle (G0/G1
Acknowledgement
This work was supported by the National Science Council of Taiwan (NSC-94-2321-B-037-006) to Lea-Yea Chuang.
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2021, South African Journal of BotanyCitation Excerpt :The acetogenins are known for their potent inhibitory effect of mitochondrial (complex I) and cytoplasmic (anaerobic) production of adenosine triphosphate (ATP), which lead to tumor cell apoptosis (programmed cell death) (McLaughlin, 2008). Annonacin, an acetogenin with a mono-tetrahydrofuran moiety, has been reported to inhibit the growth of estrogen receptor-α (ERα)-positive breast cancer cell line (MCF-7) (Ko et al., 2011; Yuan et al., 2003). Moreover, many studies have illustrated the antitumor effect of other acetogenins as well, such as bullatacin, asimicin, and trilobacin, against various cancers including gastric (Liet al., 2017), colon (Zhao et al., 1994), prostate and pancreatic cancer (Zhao et al., 1996).