In vitro evaluation of porcupine bezoar extracts as anticancer agent on A549 -A preliminary study

Porcupine bezoar (PB) was reported to possess medicinal properties in old medical manuscript. However, its potential as anticancer agent on human lung cancer cells (A549) is not yet studied. In present study, porcupine bezoar was tested to observe its ability in inhibiting cell growth of cancer cell (A549) and its cytotoxicity on Normal Human Gingival Fibroblast Cell (HGF-1). A549 cells morphology was observed after treated with bezoars for 72 hours. The ability of bezoars to induce DNA damage and apoptosis was analyzed by staining cells with Hoechst 33428(nucleus) and Rhodamine Phalloidin (f-actin). The A549 IC50 is 13.6±1.58μg/ml A549 growths was inhibited in dose-dependent pattern, but no inhibition found on normal HGF-1 cells. Treated A549 morphology shows sign of apoptosis such as DNA fragmentation, cytoplasm shrunk and vacuolation. The finding in this study suggests PB extracts able to inhibit cell growth, induce DNA damage and apoptosis, further analysis need to be done to verify the mechanism

Anticancer activity of grassy Hystrix brachyura bezoar and its mechanisms of action: An in vitro and in vivo based study

Porcupine bezoar (PB) is a calcified undigested material generally found in porcupine’s (Hystrix brachyura) gastrointestinal tract. The bezoar is traditionally used in South East Asia and Europe for the treatment of cancer, poisoning, dengue, typhoid, etc. However, limited scientific studies have been performed to verify its anticancer potential to substantiate its traditional claims in the treatment of cancers. Hence, this study was aimed at investigating the in vitro and in vivo anticancer properties of two grassy PB aqueous extract (PB-A and PB-B) using A375 cancer cell line and zebrafish model, respectively. This paper presents the first report on in vitro A375 cell viability assay, apoptosis assay, cell cycle arrest assay, migration assay, invasion assay, qPCR experimental assay and in vivo anti-angiogenesis assay using the grassy PBs. Experimental findings revealed IC50 value are 26.59 ± 1.37 μg/mL and 30.12 ± 3.25 μg/mL for PB-A and PB-B respectively. PBs showed anti-proliferative activity with no significant cytotoxic effect on normal human dermal fibroblast (NHDF). PBs were also found to induce apoptosis via intrinsic pathway and arrest cell cycle at G2/M phase. Additionally, the findings indicated its ability to debilitate migration and invasion of A375 cells. Further evaluation using embryo zebrafish model revealed LC50 = 450.0 ± 2.50 μg/mL and 58.7 ± 5.0 μg/mL for PB-A and PB-B which also exerted anti-angiogenesis effect in zebrafish. Moreover, stearic acid, ursodeoxycholic acid and pregnenolone were identified as possible metabolites that might contribute to the anticancer effect of the both PBs. Overall, this study demonstrated that PB-A and PB-B possess potential in vitro and in vivo anticancer effects which are elicited through selective cytotoxic effect, induction of apoptosis, inhibition of migration and invasion and anti-angiogenesis. This study provides scientific evidence that the porcupine bezoar do possess anti-cancer efficacy and further justifies its traditional utility. However, more experiments with higher vertebrae models are still warranted to validate its traditional claims as an anticancer agent

<i>Hystrix Brachyura</i> Bezoar Characterization, Antioxidant Activity Screening, and Anticancer Activity on Melanoma Cells (A375): A Preliminary Study

Porcupine bezoars (PBs) are masses of undigested calcareous concretions formed within the gastrointestinal tract. There are undocumented claims that PBs have antioxidant activity and can treat cancers. However, limited scientific study has been carried out to verify these traditional claims. Hence, this study was conducted to characterize the chemical profile and validate the antioxidant and anticancer activity against melanoma cells (A375). PB extract was initially subjected to Fourier-transform infrared spectroscopy (FTIR), gas chromatography&#8315;mass spectrometry (GCMS), total phenolic content (TPC), and total flavonoid content (TFC) analyses. The bioautography of antioxidant assays, namely 2,2&#8242;-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid (ABTS), 2,2-diphenyl-1-picrylhydrazy (DPPH), and &#946;-carotene was performed. An in vitro A375 cell viability assay, apoptosis assay, cell cycle arrest assay, and gene expression assay were carried out as well. The experimental finding revealed 5,10-diethoxy-2,3,7,8-tetrahydro-1H,6H-dipyrrolo[1,2-a:1&#8242;,2&#8242;-d]pyrazine, ursodeoxycholic acid, and cholest-5-en-3-ol (3 beta)-, carbonochloridate are major compounds detected in PB extract. PB extract has low phenolic content, viz. 698.7 &#177; 0.93 (&#181;g GAE/5 mg dry weight). The bioautography antioxidant assays revealed a potent antioxidant effect (ABTS &gt; DPPH &gt; &#946;-carotene), with free radical scavenging activity. Furthermore, PB extract exhibited dose- and time-dependent inhibition of cancer activity on A375 cells due to the exhibition of apoptosis via an intrinsic pathway

Porcupine bezoar exhibit cell cycle arrest through inhibiting cyclin /CDK1 complex and apoptosis mitochondria mediated dependent pathway in A549 cells

Porcupine bezoar (PB) previously reported to possess various medical properties, however there is no information on its potential anticancer effect on human cancer cells. In this study we investigate potential apoptotic effects induced by PB extracts in A549 cells and elucidate the underlying mechanism. Our study revealed that PB reduced the viability of A549 cells in dose dependent manner. PB treatment shown induction morphological changes of apoptosis feature in our phase contrast, hoechst 33342 and rhodamine phalloidin counter staining analysis. Flow cytometric analysis revealed that PB led to cell cycle arrest at G0/G1 through promoting p21 and inhibiting cyclin D/CDK1 complex gene expressions. Apoptotic gene expressions results indicated that PB mediated apoptosis of A549 cells by mitochondria pathway through receptor. Evidenced up-regulation of caspase 8 and BCL-2 interacting domain (BID) supported which resulted in Cytochrome C released. Additionally, upregulation of Bax result in down regulation of Bcl-2. The activities of caspase-8 and caspase-9 were enhanced by PB promoting caspase-3 activation, leading to DNA fragmentation. In conclusion, PB induced apoptosis occurs via mitochondria dependent pathway through receptor

Porcupine bezoar: in vitro antioxidant and anti-proliferative effects

Objectives: Porcupine bezoar (PB) is a mass of undigested organic and inorganic materials formed within the gastrointestinal tract of the porcupine. PB has been claimed to possess medicinal properties to treat different types of diseases including cancer. However, these traditional claims are yet to be scientifically ascertained and properly validated. Hence, this study was aimed to evaluate antioxidant and anticancer activities of PB through different biological assays. Design and method: Powdered PB was sonicated with double distilled water to get aqueous extract (PBA) which was initially screened for its phenolic content, flavonoid content and anti-oxidant potential using total phenolic content (TPC), total flavonoid content (TFC) and diphenl-2-picrylhydrazyl (DPPH) assays, respectively. Later on, in vitro anti-proliferative effect of PBA was evaluated against A375 (Skin Malignant Melanoma) and HGF-1 (normal cell). Results: PBA was found to contain low level of phenolic compounds and devoid of flavonoids. However, the DPPH assay showed low IC50 value indicating PBA’s potent anti-oxidant characteristic. Moreover, the PBA displayed low IC50 value and and also showed significant anti-proliferation pattern at 24, 48 and 72 hours exposures against A375 and HGF-1 cell lines. Conclusions: The results of this study suggest that PB is medicinally potent in nature due to its strong antioxidant and anti-proliferative effects and could play an important role to cure different kinds of cancers. Keywords: Porcupine bezoar, aqueous extract, TPC, TFC, DPPH, antioxidant activity, anti-proliferative acitivity, A375 and HGF-1 cell line

Anticancer activity of grassy Hystrix brachyura bezoar and its mechanisms of action: An in vitro and in vivo based study

Porcupine bezoar (PB) is a calcified undigested material generally found in porcupine’s (Hystrix brachyura) gastrointestinal tract. The bezoar is traditionally used in South East Asia and Europe for the treatment of cancer, poisoning, dengue, typhoid, etc. However, limited scientific studies have been performed to verify its anticancer potential to substantiate its traditional claims in the treatment of cancers. Hence, this study was aimed at investigating the in vitro and in vivo anticancer properties of two grassy PB aqueous extract (PB-A and PB-B) using A375 cancer cell line and zebrafish model, respectively. This paper presents the first report on in vitro A375 cell viability assay, apoptosis assay, cell cycle arrest assay, migration assay, invasion assay, qPCR experimental assay and in vivo anti-angiogenesis assay using the grassy PBs. Experimental findings revealed IC50 value are 26.59 ± 1.37 μg/mL and 30.12 ± 3.25 μg/mL for PB-A and PB-B respectively. PBs showed anti-proliferative activity with no significant cytotoxic effect on normal human dermal fibroblast (NHDF). PBs were also found to induce apoptosis via intrinsic pathway and arrest cell cycle at G2/M phase. Additionally, the findings indicated its ability to debilitate migration and invasion of A375 cells. Further evaluation using embryo zebrafish model revealed LC50=450.0 ± 2.50 μg/mL and 58.7 ± 5.0 μg/mL for PB-A and PB-B which also exerted anti-angiogenesis effect in zebrafish. Moreover, stearic acid, ursodeoxycholic acid and pregnenolone were identified as possible metabolites that might contribute to the anticancer effect of the both PBs. Overall, this study demonstrated that PB-A and PB-B possess potential in vitro and in vivo anticancer effects which are elicited through selective cytotoxic effect, induction of apoptosis, inhibition of migration and invasion and anti-angiogenesis. This study provides scientific evidence that the porcupine bezoar do possess anti-cancer efficacy and further justifies its traditional utility. However, more experiments with higher vertebrae models are still warranted to validate its traditional claims as an anticancer agent