• Asian Pacific Journal of Cancer Prevention
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• v.15 no.5
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• pp.2083-2087
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• 2014

Colorectal cancers remain to be a common cause of cancer-related death. Early-onset cases as well as those of various ethnic origins have aggressive clinical features, the basis of which requires further exploration. The aim of this work was to examine the expression patterns of $p15^{INK4b}$ and SMAD4 in colorectal carcinoma of different ethnic origins. Fifty-five sporadic colorectal carcinoma of Egyptian origin, 25 of which were early onset, and 54 cancers of Finnish origin were immunohistochemically stained with antibodies against $p15^{INK4b}$ and SMAD4 proteins. Data were compared to the methylation status of the $p15^{INK4b}$ gene promotor. $p15^{INK4b}$ was totally lost or deficient (lost in ${\geq}50%$ of tumor cell) in 47/55 (85%) tumors of Egyptian origin as compared to 6/50 (12%) tumors of Finnish origin (p=7e-15). In the Egyptian cases with $p15^{INK4b}$ loss and available $p15^{INK4b}$ promotor methylation status, 89% of cases which lost $p15^{INK4b}$ expression were associated with $p15^{INK4b}$ gene promotor hypermethylation. SMAD4 was lost or deficient in 25/54 (46%) tumors of Egyptian origin and 28/48 (58%) tumors of Finnish origin. 22/54 (41%) Egyptian tumors showed combined loss/deficiency of both $p15^{INK4b}$ and SMAD4, while $p15^{INK4b}$ was selectively lost/deficient with positive SMAD4 expression in 24/54 (44%) tumors. Loss of $p15^{INK4b}$ was associated with older age at presentation (>50 years) in the Egyptian tumors (p=0.04). These data show for the first time that $p15^{INK4b}$ loss of expression marks a subset of colorectal cancers and ethnic origin may play a role in this selection. In a substantial number of cases, the loss was independent of SMAD4 but rather associated with $p15^{INK4b}$ gene promotor hypermethylation and old age which could be related to different environmental exposures.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.12
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• pp.6239-6244
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• 2012

Background: The basal cell carcinoma (BCC) is the most common non-melanoma skin cancer (NMSK). BCC might develop because of the faulty cell cycle arrest. $p15^{INK4b}$ is a tumor suppressor gene, involved in cell cycle arrest and inactivated in most human cancers. The role of $p15^{INK4b}$ protein expression in the genesis of BCC is as yet unknown. In a previous study we showed the absence of $p15^{INK4b}$ expression in the majority of tissue microarray cores of cutaneous squamous cell carcinoma (SCCs), another type of non-melanoma skin cancer, indicating that $p15^{INK4b}$ could possibly be involved in the pathogenesis of cutaneous SCC. The aim of this study was to investigate $p15^{INK4b}$ protein expression in BCCs. Materials and Method: Protein expression of $p15^{INK4b}$ in 35 cases of BCC tissue arrays and 19 cases of normal human skin tissue was studied using an immunohistochemical approach. Results: The expression of $p15^{INK4b}$ was not significantly different in the BCC cases as compared with normal human skin (p=0.356; p>0.05). In addition, there were no significant relationship between clinicopathologic variables of patients (age and sex) and $p15^{INK4b}$ protein expression. Conclusions: Our finding may indicate that $p15^{INK4b}$ protein expression does not play a role in the genesis of BCC.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.11
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• pp.4539-4543
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• 2014

Since the epigenetic alteration in tumor cells can be reversed by the dietary polyphenol quercetin (Q) or butyrate (B) with chemopreventive activity, suggesting that Q or B can be used for chemopreventive as well as therapeutic agent against tumors. In this study the polyphenol flavonoid quercetin (Q) or sodium butyrate (B) suppressed human esophageal 9706 cancer cell growth in dose dependent manner, and Q combined with B (Q+B) could further inhibit Eca9706 cell proliferation than that induced by Q or B alone, compared with untreated control group (C) in MTT assay. The reverse expressions of global DNMT1, $NF-{\kappa}Bp65$, HDAC1 and Cyclin D1 were down-regulated, while expressions of caspase-3 and $p16INK4{\alpha}$ were up-regulated, compared with the C group in immunoblotting; the down-regulated HDAC1-IR (-immunoreactivity) with nuclear translocation, and up-regulated E-cadherin-IR demonstrated in immunocytochemistry treated by Q or B, and Q+B also displayed further negatively and positively modulated effects compared with C group. The order of methylation specific (MS) PCR of $p16INK4{\alpha}$: C>B/Q>Q+B group, while the order of E-cadherin expression level was contrary, Q+B>Q/B>C group. Thus, Q/B, especially Q+B display reverse effect targeting both altered DNA methylation and histone acetylation, acting as histone deacetylase inhibitor mediated via epigenetic-$NF-{\kappa}B$ cascade signaling.

• International Journal of Oral Biology
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• v.32 no.1
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• pp.35-43
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• 2007

Tooth loss in elderly is mainly caused by alveolar bone loss via severe periodontitis. Although the severity of periodontitis is known to be affected by age, the aging process or the genetic changes during the aging of periodontal tissue cells are not well characterized. In this study, we investigated the effect of in vitro aging on the change of gene expression pattern in periodontal fibroblasts. Gingival fibroblasts (GF) and periodontal ligament fibroblasts (PDL) were obtained from two young patients and replicative senescence was induced by sequential subcultivation. When more than 90% cells were positively stained with senescence-associated ${\beta},-galactosidase$, those cells were regarded as aged cells. In aged GF and PDL, the level of phosphorylated retinoblastoma (RB) and $p16^{INK4A}$ protein was significantly decreased and increased, respectively. However, the protein level of p53 and p21, well known senescence-inducing genes, did not increase in aged GF and PDL. Although $p27^{Kip1}$ and $p15^{INK4B}$, another cyclin-dependent kinase inhibitors, were reported to be involved in replicative senescence of human cells, they were decreased in aged GF and PDL. Because senescent cells showed flattened and enlarged cell shape and are known to have increased focal adhesion, we examined the protein level of several integrins. Aged GF and PDL showed increased protein level of integrin ${\alpha}2$, ${\alpha}v$, and ${\beta}1$. When the gene expression profiles of actively proliferating young cells and aged cells were compared by cDNA microarray of 3,063 genes and were confirmed by reverse transcription-polymerase chain reaction, 7 genes and 15 genes were significantly and commonly increased and decreased, respectively, in aged GF and PDL. Among them, included are the genes that were known to be involved in the regulation of cell cycle, gene transcription, or integrin signaling. The change of gene expression pattern in GF and PDL was minimally similar to that of oral keratinocyte. These results suggest that $p16^{INK4A}/RB$ might be involved in replicative senescence of periodontal fibroblasts and the change of gene expression profile during aging process is cell type specific.

• Journal of the East Asian Society of Dietary Life
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• v.16 no.6
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• pp.707-716
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• 2006

To make tofu (soybean curd) with cuttlefish int the cuttlefish ink diluted 20-fold was added to soymilk in the ratio of 0%, 1%, 3%, 5%, 7% and 9% there is no respective comparison here respectively and the prepared cuttlefish inky tofu samples were stored for 15 days at $4^{\circ}C$. After storage, the tofu samples were tested for yield, pH, acidity, bacterial growth, sensory evaluations and physical properties. The yields of 7%(I7) and especially 9%(I9) cuttlefish ink tofu were higher than that of the control tofu(I0). The pH was decreased, but the acidity was increased with increasing storage period. The microorganism count of I9 was the lowest during the storage period. The turbidity gradually increased until 9 days of storage and rapidly increased at 12 days of storage. In the measurement of inky tofu color, the L and b values were decreased during the storage period. In the texture analysis, the hardness, gumminess and brittleness of inky tofu were increased until 12 days of storage but then decreased. Chewiness was decreased with increasing storage period. Springiness of Il and I3 was higher than that of I0. In sensory evaluation, color was increased with increasing cuttlefish inky concentration. Sleekness of I3 was the highest. Hardness and chewiness of inky Il and I3 were the highest. Springiness, cohesiveness and softness were the highest in I3. In overall acceptability, I3 gained the highest score.

• The Journal of Korean Medicine
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• v.21 no.1
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• pp.53-61
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• 2000

Objectives: This study was camed out to examine the effect of five fractions of aqueous extract from Artemisia capillaris THUNB(ACT), on TGF, 1-induced apoptosis, cell viability, cell cycle progression and mRNA expression of apoptosis-related genes in human hepatocyte cell line HepG2. Methods: This study employed Tryphan blue exclusion assay, DNA fragmentation assay, Cpp32 protease activity assay and Quantitative RT-PCR analysis. Results: In the Tryphan blue exclusion assay, the butanol fraction of ACT with $TGF{\beta}$, l showed magnificent (Nice word, ut is it appropriate in a medical abstract\ulcorner) viability and the H2O fraction of ACT with $TGF{\beta}$, l also showed higher viability than only $TGF{\beta}$, l-treated group. DNA fragmentation assay showed that the butanol fraction and the H2O fraction carried inhibitory effects on apoptosis induction, with the butanol fraction displaying greater effects. The Cpp32 protease activity assay showed that the butanol fraction decreased Cpp32 protease activity. The H2O fraction of ACT had no significant effect on the Cpp32 protease activity. Quantitative RT-PCR showed that the butanol fraction suppressed Bax, p 15/INK4B, p21/Waf1, PAI-1 and increased Bcl-2 gene. Conclusions: The data shows that butanol fraction of ACT increases the hepatocyte viability and has the hepatocellular protective effect by the suppression of $TGF{\beta}$, l induced-apoptosis through gene regulation.

• Restorative Dentistry and Endodontics
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• v.26 no.4
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• pp.307-315
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• 2001

The application of Nd:YAG laser and irrigants to the root surface can change its surface configurations. The purpose of this study was to investigate the effects of Nd:YAG laser and irrigants on the apical seal of obturated canals. In this study, 66 single rooted teeth were randomly assigned to 4 group of 14 teeth each. 8 teeth were served us positive and negative controls. The teeth were divided into 6 groups as follows. Group A: Nd:YAG laser, 5% NaOCl + Rc-prep Group B: Nd:YAG laser, Saline Group C: 5% NaOCl + Rc-prep Group D: Saline Group E: Positive control Group F: Negative control 66 teeth were instrumented using Maillefer ProFile$^{\circledR}$ (Orifice Shapers, .04 taper, .06 taper Dentsply, Switzerland). Two of each group were selected at random, and the canal wall surfaces were examined under a SEM. 12 teeth of each group were obturated using by lateral condensation technique. Specimens were immersed in india ink for 7days, decalcified by 10% nitric acid, dehydrated by 75. 80. 85, 90, 95 and 100% alcohol in order cleared by methyl salicylate and then measured of dye penetration with stereomicroscope($\times$15 magnification) and Image Pro plus. The data were analyzed statistically by one-way ANOVA test and Duncan's Multiple Range test. The results were as follows : 1. The mean leakage was 0.128$\pm$0.376 for group A, 0.237$\pm$0.325 for group B, 0.397$\pm$0.468 for group C, 0.586$\pm$0.402 for group D, and there were statistically significant differences between group A and group D, group B and group D. (p<0.05). 2. Group A had better sealing ability than Group C, but there was statistically no significant differences. (p>0.05). 3. Group B had better sealing ability than Group D and there was statistically significant difference. (p<0.05). 4 Group A had better sealing ability than Group B, but there was statistically no significant difference. (p>0.05). 5. Group C had better sealing ability than Group D, but there was statistically no significant difference. (p>0.05). 6. As a result of observation under SEM, Smear layers were removed in Group A, B. but Smear layers were partially removed and smear plugs were remained in Group C, Smear layers were not removed in Group D. To be specially, Melting of smear layer were showed in Group C. 7. These results suggests that the laser has a potential in reducing the apical microleakage of obturated canals.

• Journal of Ginseng Research
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• v.39 no.2
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• pp.125-134
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• 2015

Background: Black ginseng (Ginseng Radix nigra, BG) refers to the ginseng steamed for nine times and fine roots (hairy roots) of that is called fine black ginseng (FBG). It is known that the content of saponin of FBG is higher than that of BG. Therefore, in this study, we examined antitumor effects against MCF-7 breast cancer cells to target the FBG extract and its main component, ginsenoside Rg5 (Rg5). Methods: Action mechanism was determined by MTT assay, cell cycle assay and western blot analysis. Results: The results from MTT assay showed that MCF-7 cell proliferation was inhibited by Rg5 treatment for 24, 48 and 72 h in a dose-dependent manner. Rg5 at different concentrations (0, 25, 50 and $100{\mu}M$), induced cell cycle arrest in G0/G1 phase through regulation of cell cycle-related proteins in MCF-7 cells. As shown in the results from western blot analysis, Rg5 increased expression of p53, $p21^{WAF1/CIP1}$ and $p15^{INK4B}$ and decreased expression of Cyclin D1, Cyclin E2 and CDK4. Expression of apoptosiserelated proteins including Bax, PARP and Cytochrome c was also regulated by Rg5. These results indicate that Rg5 stimulated cell apoptosis and cell cycle arrest at G0/G1 phase via regulation of cell cycle-associated proteins in MCF-7 cells. Conclusion: Rg5 promotes breast cancer cell apoptosis in a multi-path manner with higher potency compared to 20(S)-ginsenoside Rg3 (Rg3) in MCF-7 (HER2/ER+) and MDA-MB-453 (HER2+/ER) human breast cancer cell lines, and this suggests that Rg5 might be an effective natural new material in improving breast cancer.

• Restorative Dentistry and Endodontics
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• v.23 no.2
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• pp.561-574
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• 1998

The purposes of this study were to observe the difference in the root canal wall after hand instrumentation or engine-driven Ni-Ti instrumentation under the scanning electron microscope, and to evaluate the apical leakage provided by continuous wave of canal filling technique with or without root canal sealer and smear layer. Twenty recently extracted human maxillary anterior teeth were instrumented with K-type files or engine-driven Ni-Ti files, Quantec series 2000$^{TM}$ and irrigated with 5.25% NaOCl alone or 15% EDT A and final flush of 5.25% NaOCl. After the instrumentation and flushing, teeth were split in half with a knife and a mallet. They were then examined with a scanning electron microscope Forty-four recently extracted human maxillary anterior teeth were divided into four groups with and without smear layer and then warm vertical canal filling using System-B with or without sealer. The extent of leakage was scored after immersion in India ink for 1 week. The results were as follows: 1. No significant difference of smear layer was observed between K-type file-instrumented group and engine-driven Ni-Ti file-instrumented group. 2. A group without smear layer showed significantly less apical leakage than a group with smear layer when sealer was used for the canal filling (p<0.01). 3. There was no significant difference between a group without sealer and smear layer and a group without sealer and with smear layer (p<0.01). 4. In groups without smear layer, a group with sealer showed significantly less apical leakage than a group without sealer (p<0.01). 5. When root canals were irrigated with NaOCl alone, a group with sealer showed significantly less apical leakage than a group without sealer (p<0.01).