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.
Anthricin (Deoxypodophyllotoxin), a naturally occurring flavolignan, has well known anti-cancer properties in several cancer cells, such as prostate cancer, cervical carcinoma and pancreatic cancer. However, the effects of Anthricin are currently unknown in oral cancer. We examined the anticancer effect and mechanism of action of Anthricin in human FaDu hypopharyngeal squamous carcinoma cells. Our data showed that Anthricin inhibits cell viability in a dose- and time-dependent manner (IC50 50 nM) in the MTT assay and Live & Dead assay. In addition, Anthricin treated FaDu cells showed marked apoptosis by DAPI stain and FACS. Furthermore, Anthricin activates anti-apoptotic factors such as caspase-3, -9 and poly (ADP-ribose) polymerase (PARP), suggesting that caspase-mediated pathways are involved in Anthricin- induced apoptosis. Anthricin treatment also leads to accumulation of the pro-apoptotic factor Bax, followed by inhibition of cell growth. Taken together, these results indicate that Anthricn-induced cell death of human FaDu hypopharyngeal squamous carcinoma cells is mediated by mitochondrial-dependent apoptotic pathway. In summary, our findings provide a framework for further exploration on Anthricin as a novel chemotherapeutic drug for human oral cancer.
성숙 콩 종실에 함유되어져 있는 raffinose와 stachyose 성분은 난분해성 올리고당이며, 인체 내에서는 분해 및 흡수가 되지 않고 장내 가스나 소화불량을 일으킨다. Raffinose와 stachyose의 함량이 적은 콩 계통을 육성하기 위한 기초 정보를 얻기 위하여 59개의 유전자형을 이용하여 2년간 포장에서 재배한 후 raffinose와 stachyose의 함량을 분석하였다. Raffinose의 2년간 평균 함량은 3.353~10.271g/kg의 범위를 보였고 육종계통 Da-7에서 가장 낮았으며 PI506592 유전자원이 가장 높은 값을 보였으며 59개 유전자형에 대한 평균값은 5.035g/kg으로 나타났다. Stachyose의 2년간 평균 함량은 2.468~36.745g/kg의 범위를 보였고 육종계통 10S31에서 가장 낮았으며 PI283327 유전자원이 가장 높은 값을 보였으며 59개 유전자형에 대한 평균값은 17.222g/kg으로 나타났다. Raffinose 성분은 3.6~7.2g/kg의 범위에서 52개의 유전자형이 분포하였고, stachyose 성분에서는 12~24g/kg의 범위에서 39개의 유전자형이 분포하였다. Raffinose의 함량에서는 년차간 부의 상관관계를 보였고, stachyose의 함량에서는 년차간 정의 상관관계를 보였다. Raffinose와 stachyose의 함량간에는 정의 상관관계을 보였다.
MicroRNAs (miRNAs, miRs) are about 21-25 nucleotides in length and regulate mRNA translation by base pairing to partially complementary sites, predominantly in the 3’-untranslated region (3’-UTR) of the target mRNA. In this study, the expression profile of miRNAs was compared and analyzed for the establishment of miRNA-related odontoblast differentiation using MDPC-23 cells derived from mouse dental papilla cells. To determine the expression profile of miRNAs during the differentiation of MDPC-23 cells, we employed miRNA microarray analysis, quantitative real-time PCR (qRT-PCR) and Alizaline red-S staining. In the miRNA microarray analysis, 11 miRNAs were found to be up- or down-regulated more than 3-fold between day 0 (control) and day 5 of MDPC-23 cell differentiation among the 1,769 miRNAs examined. In qRT-PCR analysis, the expression levels of two of these molecules, miR-194 and miR-126, were increased and decreased in the control MDPC-23 cells compared with the MDPC-23 cells at day 5 of differentiation, respectively. Importantly, the overexpression of miR-194 significantly accelerated mineralization compared with the control cultures during the differentiation of MDPC-23 cells. These results suggest that the miR-194 augments MDPC-23 cell differentiation, and potently accelerates the mineralization process. Moreover, these in vitro results show that different miRNAs are deregulated during the differentiation of MDPC-23 cells, suggesting the involvement of these genes in the differentiation and mineralization of odontoblasts.
Anthocyanins are naturally occuring phytochemicals and the main components of the coloring of plants, flowers and fruits. They are known to elicit antioxidative, anti-inflammatory and cancer preventive activity. In this study, we investigated anthocyanins in black / yellow soybean seedcoats using different methods of detection - thin layer chromatography (TLC), capillary zone electrophoresis (CZE) and HPLC analysis. The anthocyanins in soybean seedcoats were extracted by five independent methods of extraction and the aglycons (anthocyanidins) of the corresponding anthocyanins were prepared by acid mediated hydrolysis. The anthocyanin / anthocyanidin in black soybean seedcoat showed characteristic TLC mobility, CZE electrophoretic retention and HPLC migration time while little of anthocyanins were detected from yellow soybean seedcoat. The extracted anthocyanins showed pH dependent retention time in CZE and spectral change in UV-Vis spectrum. HPLC analysis of the hydrolyzed extract of black soybean seedcoat identified the presence of four anthocyanidins. The major anthocyanin in black soybean seedcoat was cyanin (cyanidin-3-O-glucoside), with the relative order of anthocyanidin in cyanidin > delphinidin > petunidin > pelargonidin.
In using both hands, everyone dominantly use one hand and it is called left-handedness or right-handedness person. Measurements of grip and pinch strength provide objective indexes to represent functional integrity of the upper extremity. This study was conducted for thirty female college students(19 right-handedness and 11 lefthandedness). For assessment of the type of handedness, questionnaire was used; for grip strength, Jamar dynamometer was used; for pinch strength, Jamar pinch gauge was used. In right handedness, the grip and pinch strength of the dominant right hand was significantly higher than those of the non-dominant hand. In addition, regular exercises were shown to give influences on reduction of strength gaps between dominant and non-dominant hands. In both groups of left and right handedness, the grip and pinch strength of the dominant hand were significantly higher than those of the non-dominant hand, and regular exercises were shown to give influences on reduction of strength gaps between dominant and non-dominant hand.
Angelica decursiva has been used in Korean traditional medicine as an antitussive, an analgesic, an antipyretic and a cough remedy. However, its anti-cancer properties have not yet been well defined. In our current study, we report the cytotoxic activity and the mechanism of cell death induced by ethanol extracts of Angelica decursiva (EEAD) against the human oral cancer cell line, KB. Treatment of KB cells with EEAD induced apoptotic cell death in both a dose- and time-dependent manner as determined by MTT assay and DNA fragmentation. However, no cytotoxic effects of EEAD against human normal oral keratinocytes (HNOK) were evident. By western blot analysis, we found that apoptosis in KB cells is associated with a decrease in procaspase-7 and -9. In addition, the activation of caspase-7 was detectable in living KB cells by fluorescence microscopy. These results suggest that EEAD exhibits anti-cancer activity in KB cells via apoptosis and thus has potential as an anticancer agent in future drug development strategies.
Background : Some of invasive plants, which were introduced from foreign countries, have caused problems in Korea. Invasion of these invasive plants in the ecosystem threatens the habitat of endemic species, reducing biodiversity, and causing a disturbance in the ecological system. Hypochaeris radicata L. (Asteraceae), the most invasive plants in Korea, particularly in Jeju Island, invade farmland, and autochthonous forest, establishing monocultures and modifying the ecosystem structure. This invasive species has become a serious environmental problem because they displace the indigenous plant species. This study was conducted to evaluate the antioxidantive effects of ethanolic extracts from different parts (root, stem, seed and leaf) of the invasive exotic species Hypochaeris radicata L. Methods and Results : The aim of present study was to estimate the total phenolic and flavonoid contents and to investigate in vitro antioxidant potential of ethanolic leaf, root, seed, and stem extracts of the Hypochaeris radicata. Antioxidant activity was assessed by using 2,2-diphenyl-1-picryl-hydrazyl (DPPH) assay, reducing power activity, [2,2-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid)] ABTS+ assay and ferrous ion chelating activity. The total phenolic and flavonoid contents were also determined and expressed in gallic acid and quercetin equivalent respectively. The results of the study indicate that the ethanolic extracts of the leaf, root, seed, and stem of H. radicata posses significant scavenging activity against DPPH (21.25% for leaf, 34.98% for root, 60.76% for seed and 45.25% for stem at 250 μg/ml each) and ABTS+ radical scavenging activity (14.85% for leaf, 17.40% for root, 35.91% for seed and 24.70% for stem at 250 μg/ml each), reducing power activity (0.178 absorbance at 300 μg/ml for leaf, 0.211 absorbance at 300 μg/ml for root, 0.447 absorbance at 300 μg/ml for seed, 0.276 absorbance at 300 μg/ml for stem). The free radical scavenging and antioxidant activities may be attributed to the presence of adequate phenolic (gallic acid content is 361.92.98 μg/g in leaf, 356.59μg/g in root, 719.72 μg/g in seed and 512.08 μg/g stem) and flavonoid compounds (219.52 μg/g in leaf, 75.67μg/g in root, 281.39 μg/g in seed and 215.66 μg/g stem). This study revealed that the ethanolic extracts of both leaf, root, seed and stem of H. radicata has demonstrated significant antioxidant activity. Conclusion : In conclusion, the present study has demonstrated that Hypochaeris radicata seed ethanol extracts are rich in phenolics and have a strong antioxidant activity and a radical-scavenging action in all of the tested methods. This suggests that Hypochaeris radicata is a good source of natural antioxidants.
Soybean (Glycine max (L.) Merr.) is an important crop for protein, oil, carbohydrates, isoflavones, and many other nutrients to humans and animals. But, antinutritional factors in the raw mature soybean are exist. Raffinose and stachyose are main antinutritional factors in soybean seed. Both raffinose and stachyose are carbohydrates, belonging to the raffinose family of oligosaccharides (RFOs). RFOs are not readily digested in humans and cause flatulence or diarrhea. The objective of this research is to obtain the information on raffinose and stachyose content according to genotype and environment. A total of twenty two soybean genotypes (11 cultivars, 3 germplasms and 8 breeding lines) were selected. Each genotype was grown in the field for two years with two replications and harvested in bulk at natural maturity for two years. Content of raffinose and stachyose was detected by HPLC. The raffinose content (g/kg) of 22 genotypes was 2.68±0.21 - 5.87±2.43 in year 1 and was 3.24±0.37 - 9.05±0.16 in year 2. The stachyose content (g/kg) was 4.23±0.98 - 27.68±9.90 at year 1 and was 5.11±1.09 - 25.32±0.35 in year 2. Genotype and environment have highly significant effects on raffinose and stachyose content. Three genotypes (Da-7, 116-13, and RS-78) have low stachyose content at 5% significant level in two years. A positive correlation (R2=0.1985*) between raffinose and stachyose was observed in year 2. These informations are valuable in soybean genetics and breeding program related with raffinose and stachyose content.
Soybean proteins are widely used for human and animal feeds worldwide. The use of soybean protein has been expanded in the food industry due to their excellent nutritional benefits. But, antinutritional and allergenic factors are present in the raw mature soybean. P34 protein, referred as Gly m Bd 30K, has been identified as a predominant immunodominant allergen. The objective of this research is to identify the genetic mode of P34 protein for the improvement of soybean cultivar with a very low level of P34 protein. Two F2 populations were developed from the cross of "Pungsannamulkong" x PI567476 and "Gaechuck2ho" x PI567476 (very low level of P34 protein). Relative amount of P34 protein was observed by Western blot analysis. The observed data for the progeny of "Pungsannamulkong" and PI567476 were 133 seeds with normal content of P34 protein and 35 seeds with very low level of P34 protein (X2=1.157, P=0.20-0.30). For the progeny of "Gaechuck#1" and PI567476, the observed data were 177 seeds with normal content of P34 protein and 73 seeds with very low level of P34 protein (X2=2.353, P=0.10-0.20). From pooled data, observed data were 310 seeds with normal content of P34 protein and 108 seeds with very low level of P34 protein (X2=0.156, P=0.50-0.70). The segregation ratio (3:1) and the Chi-square value obtained from the two populations suggested that P34 protein in mature soybean seed is controlled by a single major gene. Single gene inheritance of P34 protein was confirmed in 32 F2 derived lines in F3 seeds, which were germinated from the low level of P34 protein obtained from the cross of "Pungsannamulkong" and PI567476. These results may provide valuable information to breed for new soybean line with low level of P34 protein and identification of molecular markers linked to P34 locus.
Dwarfuess and Kunitz trypsin inhibitor (KTI) protein in soybean is useful traits for basic studies. df2 and ti gene control dwarfness and the expression of Kunitz trypsin inhibitor (KTI) protein in soybean, respectively. The objective of this research was to verify genetic linkage or independent inheritance of df2 and ti loci in soybean. The F2 population was made by cross combination between "Gaechuck#2" (Df2Df2titi genotype, KTI protein absence and a normal growth type) and T210 (df2df2TiTi genotype, a dwarf growth type and KTI protein present). A total of 258 F2 seeds were analyzed for the segregation of KTI protein using SDS-PAGE. And so, 198 F2 plants were recorded for the segregation of dwarfness. The segregation ratio of 3 : 1 for Ti locus (201 Ti : 57 titi) and Df2 locus (143 Df2 : 55 df2df2) was observed. Segregation ratio of 9 : 3 : 3 : 1 (116 TiDf2: 44 Tidf2df2: 27 titiDf2: 11 titidf2df2) between df2 gene and ti gene was observed (x2 =3.53, P = 0.223). These results showed that df2 gene was inherited independently with the ti gene in soybean.
Lectin protein and Kunitz trypsin inhibitor (KTI) protein of mature soybean seed are a main antinutritional factor in soybean seed. The Le gene controls a lectin protein and Ti gene controls the KTI protein in soybean. Ti locus has been located on linkage group 9 in the classical linkage map of soybean. Position of Le locus on linkage map was not identified. Genetic relationship between Ti locus and Le locus could be useful in soybean breeding program for the genetic elimination of these factors. The objective of this study was to determine the independent inheritance or linkage between Ti locus and Le locus in soybean seed. Two F2 populations were developed from three parents (Gaechuck#1, T102, and PI548415). The F1 seeds from Gaechuck#1 (titiLeLe) x T102 (TiTilele) and Gaechuck#1 (titiLeLe) x PI548415 (TiTilele) were obtained. The lectin and KTI protein were analysed from F2 seeds harvested from the F1 plants to find independent assortment or linkage between Ti locus and Le locus. The segregation ratios of 3 : 1 for Le locus (129 Le_ : 44 lele) and Ti locus (132 Ti_ : 41 titi) and were observed. The segregation ratios of 9 : 3 : 3 : 1 (95 Le_Li_ : 34 Le_titi: 37 leleTi_ : 7 leletiti) between Le gene and Ti gene in F2 seeds were observed. This data showed that Ti gene was inherited independently with the Le gene in soybean. These results will be helpful in breeding program for selecting the line with lacking both KTI and lectin protein in soybean.