• Korean Journal of Clinical Laboratory Science
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• v.39 no.2
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• pp.128-135
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• 2007

This study was designed to investigate the correlation between the expression rate of p53 and p21 proteins by immunohistochemical staining and tumor prognostic factors including the tumor size, histological differentiation and Dukes' stage of tumor prognostic factors in colon cancer, and to acquire necessary data for the presumption of diagnosis, treatment and prognosis of colon cancer patients. From January 2000 to January 2003 at Hanyang University Guri Hospital, the paraffin blocks of 35 patients diagnosed with colon cancer whose pathologic reports were possible to review were selected. Harris hematoxylin & eosin (H&E) staining and immunohistochemical staining by ABC (Avidin Biotin Conjugate) method were performed. The histological differentiation grade and stage were classified according to the classification of the World Health Organization (WHO) and modified Dukes's stage from H&E staining. The expression rate of p53 and p21 proteins were analyzed by immunohistochemical staining. The results was analyzed statistically by SPSS (Windows version 8.0). As a result, the expression rate of p53 protein was 11.4% (4 cases) in clear differentiation, 48.6% (17 cases) in moderate differentiation, and 17.1% (6 cases) in poor differentiation. In other words, the poorer the differentiation, the higher the expression rate of p53 protein (p<0.05). The expression rate of p21 was 17.1% (6 cases) in clear differentiation, 40.0%(14 cases) in moderate differentiation, and 8.6% (3 cases) in poor differentiation, According to the progression of histological malignant degeneration, the expression rate of p21 protein decreased distinctively (p<0.05). However, the correlation between the two above mentioned proteins and the tumor-size and Dukes' stage was not of statistical significance. In the comparison of the expression rate of p53 protein with that of p21 protein, in 10 cases, p53 protein expression was positive while p21 protein expression was negative, and in 6 cases, p53 protein expression was negative whereas p21 protein expression was positive. Consequently a statistically significant inverse correlation between the expression rate of p53 protein and that of p21 protein was observed (p<0.05). In conclusion, we found a significant correlation between histological differentiation and the expression rate of p53 and p21 proteins (p<0.05), and a significant inverse correlation between the expression rate of p53 protein and that of p21 protein (p<0.05). Also, it could be confirmed that the over expression of p53 and p21 proteins is closely associated with the occurrence of colon cancer and its progress. Therefore, it is thought that this study may be greatly beneficial to the presumption of diagnosis, treatment and prognosis of colon cancer patients.

• Korean Journal of Head & Neck Oncology
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• v.21 no.2
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• pp.139-145
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• 2005

Background and Objectives: Because of squamous cell carcinoma of the head and neck undergoes a generally poor hospital course, the prognostic significance of the squamous cell carcinomas in head and neck have been evaluated to identify those features associated with aggressive biologic behavior according to the immunologic and histopathologic characteristics. Materials and Method: To assess the significance of EGFR, c-erbB-2, p21 and p53 protein in head and neck tumors and their correlation with prognostic factors, samples from 74 patients with squamous cell carcinomas of larynx, pharynx, and oral cavity were studied immunohistochemically. Results: EGFR, c-erbB-2, p21, and p53 protein were expressed 94.6%, 24.3%, 85.1%, and 55.4% in 74 cases of head and neck squamous cell carcinoma, respectively. The positive expression of EGFR was associated significantly with clinical stage and the negative expressions of p21 was associated significantly with histopathologic differentiation. There were no significant relationships between the reactivity of EGFR, c-erbB-2, p21, and p53 protein. Conclusion: The expression of EGFR, c-erbB-2, p21 and p53 protein could be related to oncogenesis, and the expression of p21 and EGFR protein can be used as a prognosticator in head and neck squamous cell carcinoma under certain limitations, but c-erbB-2 and p53 protein expression alone is not enough for evaluating prognosis of the carcinoma.

• Korean Journal of Head & Neck Oncology
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• v.15 no.2
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• pp.141-148
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• 1999

Objectives: The development of thyroid tumor has a relationship with carcinogen, oncogene and tumor suppressor gene. With aminotriazole, radioactive iodine and nitrosomethylurea as carcinogens in rat, authors investigate the incidence in type of the thyroid tumors, p21 and p53 protein expression pattern by immunohistochemical stain and the relationship between the tumors and p21-p53 protein expressions. Materials and Methods: 80 experimental animals were divided into four groups; group 1(control, no carcinogen, n=20), group 2(oral administration of aminotriazole for 36 weeks, n=20), group 3(intraperitoneal injection of 131I for one time and oral administration of aminotriazole for 36 weeks, n=20), group 4(oral administration of nitrosomethylurea for 3 days and aminotriazole for 36 weeks, n=20). After 40 weeks they were sacrificed with pathologic examination and we performed immunohistochemical staining with pan-ras monoclonal antibody for p21 protein and CMI polyclonal antibody for p53 protein with paraffin-embedded specimens. Results: 1) No tumors were observed in group I, but 38.3% of nodular goiters, 11.7% of adenomas and 50.0% of carcinomas were observed in carcinogen treated groups(group 2, 3, 4). 2) The incidence of nodular goiter, adenoma and carcinoma were 70%, 20% and 10% in group 2, 40%, 15% and 45% in group 3 and 5%, 0% and 95% in group 4. 3) p21 protein was not expressed in normal thyroid tissues but was expressed in 26.1% of nodular goiters, 42.9% of adenomas and 6.7% of carcinomas. On the other hands, p53 protein was not expressed in normal thyroid tissues, nodular goiters, adenomas and in well differentiated thyroid carcinomas by immunohistochemical stain. Conclusion: The authors suggest that aminotrizole, 131I, nitrosomethylurea can be etiologic agents in the development of thyroid tumor and the p21 protein can be expressed in the early stage and in benign condition of thyroid tumor but p53 protein is not expressed in all conditions of development in rats.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.2
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• pp.963-967
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• 2014

Previous studies have suggested anti-tumor effects of asiatic acid in some human cancer cell lines. This agent is reported to increase the levels of $p21^{WAF1/CIP1}$ in human breast cancer cell lines. However, the molecular mechanisms have not been established. Here we report that asiatic acid up-regulates $p21^{WAF1/CIP1}$ protein expression but not the level of $p21^{WAF1/CIP1}$ mRNA in HepG2 human hepatoma cells. Furthermore, we found that the asiatic acid induced increase of $p21^{WAF1/CIP1}$ protein was associated with decreased phosphorylation (ser-146) of $p21^{WAF1/CIP1}$. Knockdown of NDR1/2 kinase, which directly phosphorylates $p21^{WAF1/CIP1}$ protein at ser-146 and enhances its proteasomal degradation, increased the levels of $p21^{WAF1/CIP1}$ protein and eliminated the regulation of $p21^{WAF1/CIP1}$ stability by asiatic acid. At the same time, the expression of NDR1/2 kinase decreased during treatment with asiatic acid in HepG2 cells. Moreover, asiatic acid inhibited the proliferation of HepG2 cells, this being attenuated by knockdown of $p21^{WAF1/CIP1}$. In conclusion, we propose that asiatic acid inhibits the expression NDR1/2 kinase and promotes the stability of $p21^{WAF1/CIP1}$ protein through attenuating NDR1/2 dependent phosphorylation of $p21^{WAF1/CIP1}$ in HepG2 cells.

• Journal of Aquaculture
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• v.13 no.1
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• pp.1-7
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• 2000

The effect of various dietary protein levels on growth and body composition of young common carp raised from 56 g to 170 g in recirculating system was investigated for 15 weeks when they were fed to visual satiety three times daily. Five experiemtal diets were formulated to contain 40, 35, 30, 25 and 21% protein levels and 3.56, 3.59. 3.63, 3.66 and 3.69 kcal/g diet GE levels respectively. Mean survival rates of the fish fed the 40, 35, 30 and 25% protein diets were not different but sig-nificantly higher than that of the fish fed the 40, 35, 30 and 25% protein diets were not different but sig-nificantly higher than that of o the fish fed the 21% protein diet(P<0.05) Weight gain (g/tank) of common carp fed the 30% protein diet was the best. However weight gain of the fish fed the 25, 30, 35 and 40% protein diets were not different but significantly better than that of the fish fed the 21% protein diet. Feed efficiency ratio of the 21% protein diet was significantly lower (P<0.05) than for other groups of diets which were not different among them. Protein efficiency ratio for the 21% protein diet was significantly lower (P<0.05) than for other groups of diets which were not different among them(P>0.05) Dietary protein level had no effect on hemoglobin content in the fish(P>0.05) Crude protein contents of whole body of the fish fed the 35 and 40% protein diets were significantly higher than that of the fish fed the 21$$\mid$% or 25% protein diet(P<0.05) Body crude lipid contents of the fish fed the 21 and 25% protein diets were significantly higher than that of the fish fed the 30% or 35% protein diet. Crude ash contents of the fish fed the 35 and 40% protein diets were significantly higher than that of the fish fed the 21% or 25% protein diet(P<0.05) Moisture content of the fish fed the 35% protein diet was significantly higher than that of the fish fed the 21% protein diet(P<0.05) In considering growth performance of common carp and efficiency of diet dietary protein level could be lowered up to 25% without the reduction of young common carp production in recirculating system.

• Biomedical Science Letters
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• v.13 no.3
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• pp.213-221
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• 2007

This study was designed to investigate the correlation between the expression rate of p53 and p21 proteins, c-erbB-2 oncoprotein and $TGF-{\beta}1$ and tumor prognostic factors in colon cancer including the tumor size, histological differentiation and Dukes' stage. The expression rate of p53 protein was 11.4% (4 cases) at well differentiation, 48.6% (17 cases) at moderately differentiation, and 17.1% (6 cases) at poorly differentiation. In other words, the poorer differentiation, the higher the expression rate of p53 protein (P<0.05). The expression rate of p21 protein was 17.1% (6 cases) at well differentiation, 40.0% (14 cases) at moderately differentiation, and 8.6% (3 cases) at poorly differentiation, indicating that, as the histological malignant degeneration progressed, the expression rate of p21 protein decreased distinctively (P<0.05). However, the correlation of the above mentioned proteins with tumor size and Dukes' stage was not recognized. The expression rate of c-cerbB-2 oncoprotein was 11.4% (4 cases) at well differentiation, 54.3% (19 cases) at moderately differentiation, and 17.1% (6 cases) at poorly differentiation, indicating that the poorer differentiation, the higher expression rate of c-erbB-2 oncoprotein (P<0.05). The expression rate of $TGF-{\beta}1$ was 17.1% (6 cases) at well differentiation, 48.6% (17 cases) at moderately differentiation, and 11.4% (4 cases) at poorly differentiation. As Dukes' stage progressed, the expression rate of $TGF-{\beta}1$ was 8.6% (3 cases) in stage A, 20.0% (7 cases) in stage B, 37.1 % (13 cases) in stage C, and 11.4% (4 cases) in stage D. There was a difference in expression rates between Dukes' stages (P<0.05). In 10 cases, p53 protein was positive while p21 protein was negative, and in 6 cases, p53 protein was negative whereas p21 was positive (P<0.05). Therefore, a statistically significant inverse correlation between the expression rate of p53 protein and that of p21 protein was observed. In conclusion, since there was a signigicant correlation between histological differentiation of colon cancer and the expressions of p53 and p21 proteins and c-erbB-2 oncoprotein, and between Dukes' stage and the expression of $TGF-{\beta}1$, it was conformed that the overexpression of p53 and p21 proteins, c-erbB-2 oncoprotein, and $TGF-{\beta}1$ is closely associated with the occurrence of colon cancer and its progress. Accordingly, this study may be greatly beneficial to the presumption of diagnosis, treatment and prognosis of colon cancer patients.

• Microbiology and Biotechnology Letters
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• v.50 no.1
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• pp.63-70
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• 2022

Two small cryptic plasmids, pHG1 and pHG2, were isolated from Levilactobacillus zymae (formerly Lactobacillus zymae) GU240 and characterized. pHG1 is 1,814 bp in size with a GC content of 37.4% and contains two open reading frames. orf1 can potentially encode a protein of 101 amino acids (aa) with 99% identity with the copy number control protein of Lacticaseibacillus paracasei. orf2 can potentially encode a protein of 230 aa with 99% identity with a replication protein from multiple species. Six inverted repeats (IR I-VI) and six direct repeats (DR I-VI) were found in pHG1. pHG2 is 2,864 bp in size, with a GC content of 39.6%. pHG2 has two orfs. orf1 might encode a protein with 99% identity with the TrsL transmembrane protein. orf2 might encode a protein with 99% identity with plasmid recombination proteins from lactic acid bacteria. Both pHG1 and pHG2 may be useful as frames for constructing lactic acid bacteria-Escherichia coli shuttle vectors.

• Korean Journal of Poultry Science
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• v.30 no.2
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• pp.113-118
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• 2003

In this study, broilers were raised up to 6 weeks of age in a single room to determine if different levels of dietary protein or addition of aluminum sulfate[alum, $Al_2$(SO$_4$)$_3$ㆍ14$H_2O$] to the litter affected growth performance, production of ammonia(NH$_3$) and soluble phosphorus(SP) content of the litter.The experimental treatments consisted of six treatments in a 2x3 factorial arrangements: T$_1$＝23% protein ＋ 0.2% alum to litter; T$_2$＝21% protein ＋ 0.2% alum to litter; T$_3$＝19% protein ＋ 0.2% alum to litter; T$_4$＝23% protein ＋ no alum; T$_{5}$＝21% protein ＋ no alum; T$_{6}$＝19% protein ＋ no alum. For broiler performance, there was no effect of alum addition to the litter, but the dietary protein levels significantly affected feed intake from days 22 to 42(P<0.05) and day 0 to 42(P< 0.05), weight gain during all periods(P<0.05 or 0.01), and feed:gain from day 0 to 21(P<0.05) and day 0 to 42(P<0.05). Alum addition to the litter did not affect body weight at 21 and 42 days, but dietary protein levels has a significant effect on it at both 21(P<0.0l) and 42 days(P<0.05). Alum addition only affected ammonia production at weeks 3(P

• Molecules and Cells
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• v.28 no.5
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• pp.489-494
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• 2009

We have previously shown that the down-regulation of protein kinase CKII activity is tightly associated with cellular senescence of human fibroblast IMR-90 cells. Here, we examined the roles of p53 and $p21^{Cip1/WAF1}$ in senescence development induced by CKII inhibition using wild-type, isogenic p53-/- and isogenic p21-/- HCT116 human colon cancer cell lines. A senescent marker appeared after staining for senescence-associated ${\beta}$-galactosidase activity in wild-type HCT116 cells treated with CKII inhibitor or $CKII{\alpha}$ siRNA, but this response was almost abolished in p53- or $p21^{Cip1/WAF1}$-null cells. Increased cellular levels of p53 and $p21^{Cip1/WAF1}$ protein occurred with the inhibition of CKII. CKII inhibition upregulated p53 and $p21^{Cip1/WAF1}$ expression at post-transcriptional level and transcription level, respectively. RB phosphorylation significantly decreased in cells treated with CKII inhibitor. Taken together, this study shows that the activation of the $p53-p21^{Cip1/WAF1}$ pathway acts as a major mediator of cellular senescence induced by CKII inhibition.

• Proceedings of the Zoological Society Korea Conference
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• 1998.10b
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• pp.360-360
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• 1998

C11, a chloride-containing VK3 analog, acts as a mediator of programmed cell death in SK-Hep-1 cell lines, but its molecular mechanisms linked to cell death are not understood. In this study, we investigated the expression of p21 gene and its relationship to apoptosis induced by C11. In SK -hep-1 cells, the addition of C11 resulted in time-dependent growth suppression and DNA fragmentation characteristics of apoptosis. p21 protein was induced during this process, while the protein level of p53 was not changed at the same condition. This apoptotic cell death with p21 induction was also observed in the Hep3B cells lacking functional p53 after treatment of C11. These results suggest that C11-induced apoptosis is associated with up-regulation of p21 protein in p53-independent pathway. Next, in order to confirm whether the p53-independent p21 induction is required for C11-induced apoptosis, we introduced the p21 gene into Hep3B. Overexpression of p21 did not affect the expression of the bcl-2 gene, but DNA fragmentation and PARa cleavage were significantly increased. These data indicate that p21 is involved in C11-induced apoptosis. Although Bcl-2 has been implicated to interfere with an essential signaling molecule involved in the apoptosis pathway, its molecular mechanism and target molecule are poorly understood. To determine the effects of bcl-2 overexpression on apoptosis and to investigate whether BcI-2 interfers with the p53-independent p21 pathway, we transfected the bcl-2 expression vector into SK - Hep-1 cels. Overexpression of Bcl-2 prevented C11-induced apoptosis. Taken together, C11-induced apoptosis is regulated by p52-independent p21 pathway and bcl-2 may inhibit functional activity of p21, therebe may inhibit the C11-induced apoptosis.ptosis.