Ulva compressa Linnaeus (UCL) is a green algae seaweed that performs photosynthesis and is used as a food material in some Asian regions including Korea. It is known to be the dominant species in copper ion-contaminated seas, and many studies on copper ion resistant mechanisms have been reported. UCL is known to have an excellent antioxidant effect, but limited information is available regarding its other physiological activities. In this study, we investigated the anticancer activity of 30% prethanol extracts of Ulva compressa Linnaeus (30% PeUCL) and the underlying mechanisms of its activity on human FaDu hypopharyngeal squamous carcinoma cells. The 30% PeUCL extracts suppressed FaDu cell viability without affecting normal cells (L929), as determined by MTT and viability assays. Furthermore, the 30% PeUCL extracts induced apoptosis, as determined by DAPI staining. The 30% PeUCL extracts inhibited colony formation effectively as well as wound-healing of FaDu cells, even at noncytotoxic concentrations. In addition, 30% PeUCL extracts induced apoptosis significantly through proteolytic cleavage of caspase-3, -7, and -9, and poly (ADP-ribose) polymerase, and by downregulation of Bcl-2 and upregulation of Bax in FaDu cells, as determined by Western blot analysis. Collectively, these results suggest that the inhibitory effect of 30% PeUCL extracts on the growth of oral cancer cells, colony formation and wound-healing may be mediated by caspase- and mitochondrial-dependent apoptotic pathways in human FaDu hypopharyngeal squamous carcinoma cells. Therefore, 30% PeUCL extracts can be administered as a natural chemotherapeutic drug for the treatment of human oral cancers.

Humulus japonicus (HJ) is a widely used herbal medicine for pulmonary tuberculosis, hypertension, leprosy, and venomous wounds in Asia, particularly in China. Although HJ has certain physiological activities, such as longitudinal bone growth, antioxidation and alleviation of rheumatism, its anticancer activities, other than in colorectal and ovarian cancer, are yet to be studied. In this study, we investigated the anti-cancer activity and mechanism of methanol extracts of HJ (MeHJ) against human FaDu hypopharyngeal squamous carcinoma cells. MeHJ suppressed FaDu cell viability without affecting normal cells (L929), which was demonstrated using the MTT and Live & Dead assays. Furthermore, MeHJ effectively inhibited colony formation of FaDu cells, even at non-cytotoxic concentrations, and significantly induced apoptosis through the proteolytic cleavage of caspase-9, -3, -7, poly (ADP-ribose) polymerase and through the downregulation of BCL-2 and upregulation of BAX in FaDu cells, as determined by DAPI staining, flow cytometry, and western blot analyses. Collectively, these findings suggest that the inhibitory effects of MeHJ on the growth and colony formation of oral cancer cells may be mediated by caspase- and mitochondrial-dependent apoptotic pathways in human FaDu hypopharyngeal squamous carcinoma cells. Therefore, MeHJ has the potential to be used as a natural chemotherapeutic drug against human oral cancer.

Nypa fruticans Wurmb (NFW) contains a large amount of phenolic acid and flavonoids, and is popular as a superfood in Myanmar. NFW has various biological activities, such as anti-inflammatory, anti-oxidant, and neuroprotective properties; however, the anti-cancer effect of NFW have not been reported. In this study, we investigated the anticancer activity of water extracts of NFW (WeNFW) and the underlying mechanism in human FaDu hypopharyngeal squamous carcinoma cells. The WeNFW inhibited FaDu cell growth in a dose-dependent manner without affecting normal cells (L929), as determined by an MTT assay and Live and Dead assay. In addition, the concentrations of WeNFW without cytotoxicity (0.025, 0.05, and 0.1 mg/mL) inhibited wound healing and colony formation. Furthermore, WeNFW significantly induced apoptosis through the proteolytic cleavage of caspase-3 and -9, poly (ADP-ribose) polymerase, and downregulation of Bcl-2 and upregulation of Bax in FaDu cells, as determined by DAPI staining, FACS analysis, and western blot analysis. Taken together, these results suggest that WeNFW exhibits potent anti-cancer effects by suppressing the growth of oral cancer cells, wound healing and colony formation activity. Via mitrochondrial-dependent apoptotic pathways in human FaDu hypopharyngeal squamous carcinoma cells. Therefore, WeNFW can provide a natural chemotherapeutic drug for oral cancer in humans.

Asarum sieboldii Miq. (Aristolochiaceae) is a perennial herbaceous plant and has been used as traditional medicine for treating diseases, cold, fever, phlegm, allergies, chronic gastritis, and acute toothaches. Also, it has various biological activities, such as antiallergic, antiinflammatory, antinociceptive, and antifungal. However, the anticancer effect of A. sieboldii have been rarely reported, except anticancer effect on lung cancer cell (A549) of water extracts of A. sieboldii . This study investigated the anticancer activity of methanol extracts of A. sieboldii (MeAS) and the underlying mechanism in human FaDu hypopharyngeal squamous carcinoma cells. MeAS inhibited FaDu cells grown dose-dependently without affecting normal cells (L929), as determined by 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyl tetrazolium bromide and live and dead assay. In addition, concentration of MeAS without cytotoxicity (0.05 and 0.1 mg/mL) inhibited migration and colony formation. Moreover, MeAS treatment significantly induced apoptosis through the proteolytic cleavage of caspase-3, -7, -9, poly (ADP-ribose) polymerase, and downregulation of Bcl-2 and upregulation of Bax in FaDu cells, as determined by fluorescence-activated cell sorting analysis, 4`6-diamidino- 2-phenylindole stain, and western blotting. Altogether, these results suggest that MeAS exhibits strong anticancer effects by suppressing the growth of oral cancer cells and the migration and colony formation via caspase- and mitochondrial-dependent apoptotic pathways in human FaDu hypopharyngeal squamous carcinoma cells. Therefore, MeAS can serve as a natural chemotherapeutic for human oral cancer.

Acacetin, which is present in damiana (Turnera diffusa ) and black locust (Robinia pseudoacacia ), has several pharmacologic activities such as antioxidant, anti-inflammatory, and anti-proliferative effects on cancer cells. However, the effect of acacetin on head and neck cancers has not been clearly established. This study aimed to examine the effects of acacetin on cell growth and apoptosis induction in FaDu human pharyngeal carcinoma cells. These were investigated by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay, Live/Dead cell assay, 4′,6-diamidino-2-phenylindole dihydrochloride staining, caspase-3 and caspase-7 activation assay, and immunoblotting in FaDu cells. Acacetin induced FaDu cell death in a dose-dependent manner, with an estimated IC50 value of 41.9 µM, without affecting the viability of L-929 mouse fibroblasts as normal cells. Acacetin treatment resulted in nuclear condensation in the FaDu cells. It promoted the proteolytic cleavage of procaspase-3, -7, -8, and -9 with increasing amounts of the cleaved caspase isoforms in FaDu cells. Acacetin-induced apoptosis in FaDu cells was mediated by the expression of Fas and activation of caspase-8, caspase-3, and poly (ADP-ribose) polymerase. Immunoblotting showed downregulation of the anti-apoptotic mitochondrial proteins Bcl-2 and Bcl-xL, but upregulation of the mitochondria-dependent pro-apoptotic proteins Bax and Badin FaDu cells after acacetin treatment. These findings indicate that acacetin inhibits cell proliferation and induces apoptotic cell death in FaDu human pharyngeal carcinoma cells via both the death receptor-mediated extrinsic apoptotic pathway and the mitochondria-mediated intrinsic apoptotic pathway.

Ficus carica L. (fig ) is one of the first cultivated crops and is as old as humans. This plant has been extensively used as a traditional medicine for treating diseases, such as cough, indigestion, nutritional anemia, and tuberculosis. However, the physiological activity of fig leaves on oral cancer is as yet unknown. In this study, we investigated the anticancer effect of methanol extracts of Ficus carica (MeFC) and the mechanism of cell death in human FaDu hypopharyngeal squamous carcinoma cells. MeFC decreased the viability of oral cancer (FaDu) cells but did not affect the viability of normal (L929) cells, as determined by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay and Live and Dead assay. In addition, MeFC induced apoptosis through the proteolytic cleavage of procaspase-3, -9, poly (ADP-ribose) polymerase (PARP), downregulation of Bcl-2, and upregulation of Bax, as determined by 4′,6-diamidino-2-phenylindole dihydrochloride staining and western blot analysis. Moreover, a concentration of MeFC without cytotoxicity (0.25 mg/mL) significantly suppressed colony formation, a hallmark of cancer development, and completely inhibited the colony formation at 1 mg/mL. Collectively, these results suggest that MeFC exhibits a potent anticancer effect by suppressing the growth of oral cancer cells and colony formation via caspase- and mitochondrial-dependent apoptotic pathways in FaDu human hypopharyngeal squamous carcinoma cells. Therefore, the methanol extract of Ficus carcica leaves provide a natural chemotherapeutic drug for human oral cancer.

Trifolium pratense leaves (red clover) has been used in Oriental and European folk medicine for the treatment of whooping cough, asthma, and eczema, and is now being used to treat and alleviate the symptoms, such as hot flushes, cardiovascular health effects that occur in postmenopausal women. However, relatively little scientific data is available on the physiological activity of this plant. Therefore, in this study, we investigated the anti-cancer activity of T. pratense leaves using methanol extract of T. pratense leaves (MeTP) on human FaDu hypopharyngeal squamous carcinoma cells. MeTP inhibited the viability of FaDu cells by inducing apoptosis through the cleavage of procaspase- 3, -7, and -9 and poly (adenosine diphosphate ribose-ribose) polymerase (PARP), downregulation of Bcl- 2, and upregulation of Bax, as determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, Live & dead assay, 4’6-diamidino-2-phenylindole stain, fluorescence-activated cell sorting analysis, and Western blot analysis. In addition, colony formation was slightly inhibited when FaDu cells were treated with a non-cytotoxic concentration (0.125 mg/mL) of MeTP and almost completely inhibited when cells were treated with 0.25 mg/mL MeTP. Collectively, these results indicate that MeTP induced cell apoptosis via caspase- and mitochondrial-dependent apoptotic pathways, and inhibited colony formation of cancer cells in FaDu human hypopharyngeal squamous carcinoma cells. These findings suggest MeTP should be considered for clinical development as a chemotherapeutic option in oral cancer.

Codium fragile (Suringar) Hariot is an edible green seaweed that belong to the Codiaceae family and has been used in Oriental medicine for the treatment of enterobiasis, dropsy, and dysuria. Methanol extract of codium fragile has anti-oxidant, anti-inflammatory and anti-cancer properties, although the anti-cancer effect on oral cancer has not yet been reported. In this study, we investigated the anti-cancer activity and the mechanism of cell death by methanol extracts of Codium fragile (MeCF) on human FaDu hypopharyngeal squamous carcinoma cells. Our data showed that MeCF inhibits cell viability in a dose-dependent manner, and markedly induced apoptosis, as determined by the MTT assay, Live/Dead assay, and DAPI stain. In addition, MeCF induced the proteolytic cleavage of procaspase -3, -7, -9 and poly(ADP-ribose) polymerase(PARP), and upregulated or downregulated the expression of mitochondrial-apoptosis factor, Bax(pro-apoptotic factor), and Bcl-2(anti-apoptotic factor), . Futhermore, MeCF induced a cell cycle arrest at the G1/S phase through suppressing the expression of the cell cycle cascade proteins, p21, CDK4, CyclinD1, and phospho-Rb. Taken together, these results indicated that MeCF inhibits cell growth, and this inhibition is mediated by caspase- and mitochondrial-dependent apoptotic pathways through cell cycle arrest at the G1/S phase in human FaDu hypopharyngeal squamous carcinoma cells. Therefore, methanol extracts of Codium fragile can be provided as a novel chemotherapeutic drug due to its growth inhibition effects and induction of apoptosis in human oral cancer cells.

Ficus carica L. (common fig), one of the first plants cultivated by humans, originated in the Mediterranean basin and currently grows worldwide, including southwest Asia and South Korea. It has been used as a traditional medicine for treatment of metabolic, cardiovascular, and respiratory diseases as well as hemorrhoids and skin infections. Its pharmacological properties have recently been studied in detail, but research on the anti-cancer effect of its latex has been only been studied on a limited basis on several cell lines, such prostate cancer, breast cancer, and leukemia. In this study, we investigated the anti-cancer activity of the latex of Ficus carica L.and its underlying mechanism in FaDu human hypopharynx squamous carcinoma cells. (See Ed. note above) We confirmed through SDS-PAGE analysis and gelatinolytic activity analysis that the latex of Ficus carica contains cysteine protease ficin. Our data showed that the latex inhibited cell growth in a dose-dependent manner. In addition, the latex treatment markedly induced apoptosis in FaDu cells as determined by FACS analysis, elevated expression level of cleaved caspase-9, -3 and PARP (poly (ADP-ribose) polymerase), and. increased the expression of Bax (pro-apoptotic factor) while decreasing the expression of Bcl-2 (anti-apoptotic factor). Taken together, these results suggested that latex containing the ficin inhibited cell growth and induced apoptosis by caspase and the Bcl-2 family signaling pathway in FaDu human hypopharynx squamous carcinoma cells. These findings point to the potential of latex of Ficus carica to provide a novel chemotherapeutic drug due to its growth inhibition effects and induction of apoptosis in human oral cancer cells.

Anthriscus sylvestris (L.) Hoffm. is a perennial herb found widely distributed in various regions of Korea, Europe, and New Zealand. The root of A. sylvestris have been extensively used in the treatment for antitussive, antipyretic, cough remedy in Oriental medicine, but the physiologically active function of the leaf of A. sylvestris is as yet unknown. In this study, we investigated the anti-cancer activity and the mechanism of cell death of water extracts of leaf of Anthriscus sylvestris (WELAS), on human FaDu hypopharyngeal squamous carcinoma cells. Our data showed that WELAS treatment inhibited cell viability in a concentration- and time-dependent manner. In addition, the treatment of WELAS markedly induced apoptosis in FaDu cells, as determined by the viability assay, DAPI stain and FACS analysis. WELAS also increased the proteolytic cleavage of procaspase-3, -9 and PARP (poly(ADP-ribose) polymerase). In addition, exposure to WELAS decreased the expression of Bcl-2 (an anti-apoptotic factor), but increased the expression of Bax (a pro-apoptotic factor), suggesting that mitochondria-dependent apoptotic pathways are mediated in WELAS-induced apoptosis. Taken together, these results indicate that water extracts of leaf of A. sylvestris inhibits cell growth and induces apoptosis via the mitochondrial-dependent apoptotic pathway in FaDu human hypopharyngeal squamous carcinoma cells. Therefore, we propose that the water extracts of leaf of A. sylvestris is a novel chemotherapeutic drug, having growth inhibitory properties and induction of apoptosis in human oral cancer cells.

The purpose of this study is to evaluate the quality characteristics of blanched Capsella Bursa-pastoris with quick freezing, and thawing methods. Blanched Capsella bursa-pastoris was packed by vacuum packing (VP) and air containing packing (AP). After then, they were frozen at -20, -40, and 60℃ (immersion freezing). Then, they were stored at -20℃ during 1 week. They were thawed with normal (room temperature), running water and high frequency method. The pH and color of Capsella bursa-pastoris were not different before and after freezing. Likewise, total bacterial counts were shown without variation among them. The amount of drip from thawed Capsella bursa-pastoris was generally more in VP than AP. The cutting intensity of immersion freezing wes higher than that of other freezing methods (P<0.05). Therefore, we selected AP, immersion freezing and high frequency thawing methods for the high quality according to above results

The aim of this study is to assess the replacement effect of black chokeberry (Aronia melanocarpa) used in red-pepper paste seasoning with vinegar. Red-pepper paste seasoning with aronia and vinegar was prepared by mixing red pepper paste, aronia juice or vinegar, sugar and honey, and then aronia juice (J) was added to 0% (C), 50% (A1) and 100% (A2) by replacing the vinegar. As a result, the brightness of the prepared gochujang was decreased with increasing the level of the aronia juice. Total sugar and salinity of C, A1, and A2 showed a significant increase (p<0.05) in compared with that of aronia juice, 24.42 mg/mL and 1.11%, respectively. Also, pH level showed significantly increase with increasing the content of the juice (p<0.05). However, total acidity was decreased with increasing the level of the juice (p<0.05) and A2 (6.55%) especially was lower than the juice, 8.06%. Meanwhile, the total polyphenols were increased with increasing the content of the juice at gochujang samples (p<0.05) and the polyphenols content of the aronia juice was the highest at all samples (p<0.05). The total anthocyanins were also increased with increasing the level of the juice (p<0.05). Antioxidant activity measured by DPPH and ABTS radical scavenging rate was the highest at the aronia juice (p<0.05). Consequently, the result of antioxidant activity of red-pepper paste seasoning with aronia suggested that aronia can be used as a substitute of vinegar in red-pepper paste seasoning with vinegar.

Isomaltooligosaccharides were prepared from puffed rice flour by a One-step processing method under different conditions, and their physicochemical properties were evaluated. Sample was prepared by allowing puffed rice flour to pass through a 50 mesh standard sieve and segregated into experimental groups (PR15-60, PR20-60, PR15-65, and PR20-65) according to the substrate concentrations (15%, 20% w/v) and reaction temperatures (60 , 65 ). Enzyme reaction of puffed rice flour and water mixture was performed for 0, 1, 6, 24, and 48 h using commercial enzymes (Maltogenic amylase, Promozyme D2 and Transglucosidase; amount: 0.5% (w/v) to obtain the corresponding isomaltooligosaccharides. The reducing sugar content, dextrose equivalent, and total soluble solids of the reactants increased with increasing reaction time. The carbohydrate composition and amount of isomaltooligosaccharides with degree of polymerization (DP) 2 to DP 7 in the rice flour were examined by HPLC with an evaporative light scattering detector. Because of the high carbohydrate composition, the PR20-65 group showed the highest isomaltooligosaccharide content after 6 h of reation (138.47 mg/mL). After 24 h of reaction, the amount of isomaltooligosaccharides (DP2-DP7) and the isomaltooligosaccharides/total carbohydrate ratio in this group were 135.00 mg/mL and 68.04%, respectively.

약호박을 페이스트로 소재로 활용하기 위한 가열조건을 확인하기 위하여 증숙 후 고압가열 처리 시간을 달리한 약호박 페이스트를 제조하여 색도, 가용성 고형분, 유리당, 유기산 및 관능적 특성을 확인하였다. 그 결과, 색도는 증숙처리 후 L, b 값이 유의적으로 증가하였으나, a 값은 증숙과 고압가열 시간이 증가함에 따라 유의적으로 감소하였으며 페이스트의 고압가열 시간이 증가함에 따라 L, a 및 b value 모두 유의적으로 감소하였다. 가용성 고형분은 증숙 처리 페이스트가 4.13 °Brix으로 생약호박 대비 유의적으로 증가하였으나, 고압가열 처리 시간이 증가함에 따라 3.97-3.80 °Brix으로 유의적으로 감소하는 경향을 보였다(p<0.05). 유리당은 증숙 처리 후 sucrose가 추가적으로 검출되었으며, HHP0 실험군의 fructose, glucose 및 sucrose 함량이 가열조건별 약호박 페이스트 중 유의적으로 가장 높은 값을 나타내었고(p<0.05), 고압가열시간이 증가함에 따라 fructose의 값은 유의적으로 감소하였고, glucose는 HHP40 실험군이 유의적으로 가장 낮은 값을 보였으며, sucrose는 HHP60 실험군이 유의적으로 가장 낮은 값을 보였다. 총 유리당 함량은 HHP0 실험군이 1,413.36 mg/100 g으로 유의적으로 가장 높았으며, 고압가열처리가 40분 이상 지속 시, 총 유리당 함량이 1,068.89 mg/100 g으로 감소하였다. 비휘발성 유기산은, 증숙 및 고압가열 처리 시 succinic acid가검출되지 않았고, malic acid는 생약호박에 비해 유의적으로 증가하여 405.0-428.9 mg/100 g의 범위를 보인 반면 fumaric aicd는 큰 폭으로 감소했다. 총 유기산 함량은 유의적인 차이가 나타나지 않았으나 증숙 처리에 의해 그 함량이 증가하는 경향을 보였다. 고압가열 처리에 따른 약호박 페이스트의 관능적 특성차이 검사 결과 40분간 고압가열 처리한 HHP40 실험군의 향, 단맛, 감칠맛 특성이 가장 높게 나타났으나 유의적인 차이는 없었고, 관능적 기호도 검사 결과 40분간 고압가열 처리한 HHP40 실험군의 향, 단맛, 감칠맛, 전반적기호도 점수가 가장 높았으나 모두 유의적인 차이는 없는 것으로 나타났다. 위 모든 결과를 종합하여 볼 때, 20분간 증숙 처리한 약호박 페이스트의 가용성 고형분, 총 유리당 함량, 총 유기산 함량이 가장 높았고, 고압가열 처리와 그 지속시간에 따른 관능적 특성이 크게 개선되지 않은 것으로 보아 약호박을 이용한 소재를 제조하기 위한 장시간의 고압가열 처리는 불필요할 것으로 사료된다. 또한, 약호박은 단호박과 달리 가용성 고형분이 낮고, 단맛 특성, 단맛 기호도 및 전반적 기호도 점수가 낮은 소재인 것으로 나타나, 기호도를 향상시키기 위해 감미료 등 부재료를 첨가하여 페이스트로 제조 및 이의 품질특성에 대한 추가적인 연구가 필요할 것으로 보인다.