Graviola (Annona muricata) Exerts Anti-Proliferative, Anti-Clonogenic and Pro-Apoptotic Effects in Human Non-Melanoma Skin Cancer UW-BCC1 and A431 Cells In Vitro: Involvement of Hedgehog Signaling

Abstract

Non-melanoma skin cancers (NMSCs) are the leading cause of skin cancer-related morbidity and mortality. Effective strategies are needed to control NMSC occurrence and progression. Non-toxic, plant-derived extracts have been shown to exert multiple anti-cancer effects. Graviola (Annona muricata), a tropical fruit-bearing plant, has been used in traditional medicine against multiple human diseases including cancer. The current study investigated the effects of graviola leaf and stem extract (GLSE) and its solvent-extracted fractions on two human NMSC cell lines, UW-BCC1 and A431. GLSE was found to: (i) dose-dependently suppress UW-BCC1 and A431 cell growth, motility, wound closure, and clonogenicity; (ii) induce G0/G1 cell cycle arrest by downregulating cyclin/cdk factors while upregulating cdk inhibitors, and (iii) induce apoptosis as evidenced by cleavage of caspases-3, -8 and PARP. Further, GLSE suppressed levels of activated hedgehog (Hh) pathway components Smo, Gli 1/2, and Shh while inducing SuFu. GLSE also decreased the expression of pro-apoptotic protein Bax while decreasing the expression of the anti-apoptotic protein Bcl-2. We determined that these activities were concentrated in an acetogenin/alkaloid-rich dichloromethane subfraction of GLSE. Our data identify graviola extracts and their constituents as promising sources for new chemopreventive and therapeutic agent(s) to be further developed for the control of NMSCs.

Antiproliferative Activity, Proapoptotic Effect, and Cell Cycle Arrest in Human Cancer Cells of Some Marine Natural Product Extract

“…Each of Pg E and Ha E was found to in vitro inhibit cell proliferation and induce apoptosis in HepG2 , HCT-116 , and MCF-7 cancer cells, while Sj SG in vitro inhibited angiogenesis and osteoclastogenesis and increased cytotoxicity [ 31 ]; Am C and Aa C also in vitro inhibited cell proliferation and induced apoptosis in S180 cancer cells [ 32 ], HP E in vitro induced apoptosis on CLLB s [ 33 ], SK-MEL-2 in vitro inhibited SK-MEL-2 proliferation and metastasis [ 34 ], and PE in vitro inhibited S180 and hepatoma proliferation and increased apoptosis and antiangiogenic activity [ 25 , 26 ]. However, present in vitro anticancer effects of Pg E or Ha E were supported by the downregulation of CDK2 , upregulation of PARP in HepG2 , HCT-116 , and MCF-7 cancer cells, and Bcl-2 downturn, upregulation of Bax and caspase-3 , induction of G 0 /G 1 cell cycle arrest, and induction of apoptosis at pre G 1 phase in HepG2 cancer cell line, while other previously investigated sea cucumber anticancer effects were supposed to be as a result of Bcl-2 , STAT3 , and MMP-9 downregulation [ 34 ].…”

“…The reagents SRB, DMSO, and doxorubicin were purchased from Sigma-Aldrich, St. Louis, USA, while FBS was purchased from HyClone (Pittsburgh, PA, USA) and PSA was obtained from Mediatech Inc. (Herndon, VA, USA). Cancer cells were cultured in DMEM supplemented with 5% heat-inactivated FBS and 1% PSA at 37°C in a 5% CO 2 incubator [ 26 ].…”

“…The blot was blocked in blocking buffer (7% nonfat dry milk/1% Tween 20; in 20 mmol/L TBS (pH 7.6)) for 1 h at room temperature, incubated with the appropriate monoclonal or polyclonal primary antibody in blocking buffer for 2 h at room temperature or overnight at 4°C, followed by incubation with an appropriate secondary antibody HRP conjugate. The blots were exposed to enhanced chemiluminescence (Thermo Scientific Pierce, Rockford, IL) and subjected to autoradiography using a Bio-Rad (Hercules, CA) imaging system [ 26 , 28 ]. The digitalized scientific software program Quantity One (Bio-Rad) was used for the analysis of the Western blot to assess the densitometric measurements of the bands, and treatment protocol was carried out at least three times as well as each protein expression was analyzed three times with analogous results [ 27 , 28 ].…”

Role and Therapeutic Targeting of the PI3K/Akt/mTOR Signaling Pathway in Skin Cancer: A Review of Current Status and Future Trends on Natural and Synthetic Agents Therapy

“…BCC arises in the basal epidermal cell layers and constitutes up to 80% of skin cancers and nearly a third of all cancers diagnosed in the United States [157,158,159] with an incidence rate of up to 5% per year generating a total cost of about $400 million/year [160]. BCC is usually not life threatening, but if left untreated it can cause loss of function, and disfiguration [161,162,163,164]. The morphological features of BCC include the presence of a group of tumors in the dermal layer of the skin that are composed of cells having cellular components similar to undifferentiated basal epidermal cells.…”

Novel extracts from Callyspongia siphonella and Negombata magnifica sponges from the Red Sea, induced antiproliferative and proapoptotic activity in HepG-2, MCF-7, and Caco-2 cancer cell lines

“…DMSO was purchased from (Sigma-Aldrich, St. Louis, USA), while FBS was purchased from Hyclone (Pittsburgh, PA, USA) and PSA was obtained from Mediatech Inc. (Herndon, VA, USA). Cancer cells were cultured in DMEM supplemented with 5% heat inactivated FBS and 1% PSA at 37°C in 5% CO 2 incubator (Chamcheu et al, 2018). Similarly, HFB-4 cells were maintained in RPMI-1640 medium (Thermofisher Scientific Co., Waltham, MA, USA) supplemented with 10% heat-inactivated FBS, 100 U/ml penicillin, and 100 U/ml streptomycin.…”