• Tuberculosis and Respiratory Diseases
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• v.63 no.3
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• pp.251-260
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• 2007

Purpose: An imbalance of cell proliferation and cell apoptosis is an important mechanism in carcinogenesis. Capase 3, survivin and p53 have been identified as important members of the apoptotic related proteins. This study evaluated the proliferating cell nuclear antigen(PCNA), apoptosis, apoptotic related protein such as capase 3, survivin and p53 using urethane-induced mouse lung carcinogenesis, which provides reproducible steps from hyperplasia to adenocarcinoma. Methods: Urethane was administered to the ICR mice through an intra-peritoneal injection, The mice were sacrificed at 5, 15, and 25 weeks after urethane intervention. The sequential morphological changes and immunohistochemical expression of PCNA, apoptosis, capase 3, survivin, and p53 were examined during mouse lung carcinogenesis. Results: During carcinogenesis, the sequential histological changes were observed from hyperplasia of type II pneumocytes, to anadenoma, and ultimately to an overt adenocarcinoma. The PCNA Labeling index (LI) was 9.6% in hyperplasia, 23.2% in adenoma, and 55.7% in adenocarcinoma, respectively. The apoptotic LI was 0.24% in hyperplasia, 1.25% in adenoma, and 5.27% in adenocarcinoma. A good correlation was observed between the PCNA LI and apoptotic LI. The expression of caspase 3 was remarkable- i.e., 46.7% in adenocarcinoma, in contrast to 15% in hyperplasia and 16% in adenoma. Survivin was detected weakly in the alveolar hyperplasia and showed an increasing expressional pattern in adenoma and adenocarcinoma. p53 expression was detected only in the adenocarcinoma lesions with an expression rate of 13.3%. The level of caspase 3 expression correlated with the increase in the apoptotic index. The positive expression of caspase 3 was associated with an increased apoptotic index. Conclusions: These results suggest that the PCNA LI and apoptotic LI might be useful markers for evaluating the risk of a malignant transformation. In addition, caspase, survivin and p53 might play a role in the early and late steges of urethane-induced mouse lung carcinogenesis.

• Journal of Animal Science and Technology
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• v.46 no.4
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• pp.585-592
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• 2004

Our previous studies demonstrated that relaxin in concert with estrogen promotes development of the mammary parenchyma during the last third of gestation in gilts, and the specific relaxin-binding sites were present in the mammary gland. This study was conducted to determine if relaxin-binding sites in the mammary gland were functional relaxin receptors. Three cycling cross-bred gilts were bilaterally ovariectomized on day 0 of the experiment. Beginning on day 15 and continuing through day 29 post-surgery, the gilt received an im. injection of estradiol benzoate at 12-hr intervals. Beginning on day 22 post-surgery, highly purified porcine relaxin was administered(lug/hr) into the left fourth mammary gland from the anterior end via miniature osmotic pump. Physiological saline was administered to the right fourth mammary gland. The gilt was sacrificed on day 29 post-surgery and histological characteristics of the mammary parenchyma were examined. The mammary glands treated locally with saline showed little, if any, lobulo-alveolar development, whereas the mammary glands treated with relaxin showed not only marked lobulo-alveolar development but also prominent secretions in the alveoli. The saline-treated glands were characterized by relatively dense and highly organized collagen fiber bundles. Whereas, in the relaxin-treated mammary glands, collagen fiber bundles were dispersed and loosely organized. In conclusion, relaxin-binding sites in the mammary gland are functional relaxin receptors and relaxin acts directly on the pig mammary gland to promote development of the alveoli and remodeling of the extracellular matrix.

• Journal of Life Science
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• v.28 no.2
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• pp.141-147
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• 2018

This study aimed to analyze the contents of rutin, procyanidin B3, quercetin, and kaempferol, known to have antioxidant, anti-inflammatory, and anti-carcinogenic effects, among the polyphenol types contained in grape pruning stem extracts (GPSE). It utilized grape stems discarded after harvest to measure the effects of GPSE on skin moisture, inhibition of skin cell proliferation, and anti-inflammatory activity on the damaged skin of HR-1 mice induced with ultraviolet B (UVB), and to verify the applicability of GPSE as a material for functional food and functional cosmetics. The polyphenol was extracted from grape pruning stems with 80% EtOH, and then the extract was used while storing at $-20^{\circ}C$, after filtering, concentrating, and freeze-drying it. The content of an active ingredient of GPSE was analyzed using high performance liquid chromatography (HPLC). From 53 kg of the grape pruning stem specimen, 2.34 kg of the EtOH fraction extracts were extracted to achieve a 4.42% yield ratio. Analysis of the active ingredients showed 0.28 mg/g of procyanidin B3, 12.81 mg/g of rutin, 0.51 mg/g of quercetin, and 8.24 mg/g of kaempferol. After UVB irradiation on the dermis, to confirm the degree of inhibition of collagen synthesis, we examined the protein expression of MMP-9 using immunohistochemical staining. The results of this study confirm the existence of active polyphenol types, such as rutin, kaempferol, quercetin, and procyanidin B3, in GPSE. Moreover, the study found that GPSE has anti-collagenase effects and it decreases the effects of UV damage on skin barrier function. GPSE is a functional ingredient with a potential for skin protection effects, and it has high utilization potential as an ingredient for functional cosmetics.

• Development and Reproduction
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• v.6 no.2
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• pp.131-139
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• 2002

Since GnRH and its receptor genes are expressed in the ovary, it has been suggested that ovarian GnRH might be involved in the regulation of ovarian function and the apoptosis of ovarian cells. However, it was not known well on the expression and function of GnRH and its receptor in the corpus luteum. The present study was undertaken to investigate whether GnRH and its receptor are expressed in luteal cells and GnRH has any effect on the apoptosis of luteal cells. Luteal cells obtained from the pregnant rats were cultured and stained for GnRH and its receptor proteins. Cultured luteal cells showed distinct immunoreactivity against both anti-GnRH and anti-GnRH receptor antibodies. In addition, the presence of GnRH receptor protein in cultured cells was confirmed by Western blot analysis. To investigate the effect of GnRH on the apoptosis of luteal cells, luteal cells were cultured in the presence of 10$^{-6}$ M GnRH-agonist(GnRH-Ag) for 3, 8, and 12h. TUNEL assay showed that the number of cells undergoing apoptosis increased 12h after culture(P<0.05). DNA fragmentation analysis confirmed the results such that the cells treated for 12h showed the greatest increase of fragmentation(p<0.05). Further, Western blot analysis of cytochrome c in the mitochondrial and cytoplasmic fractions of the luteal cells showed that GnRH-Ag treatment increased the content of cytochrome c in cytoplasm. These results demonstrate that the luteal cells express GnRH and its receptor and GnRH-Ag treatment induces apoptosis of the luteal cells via mitochondrial release of cytochrome c. The present study suggest that the releasing of cytochrome c from mitochondria might be involved in the luteal cell apoptosis induced by GnRH-Ag.

• Childhood Kidney Diseases
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• v.2 no.2
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• pp.104-109
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• 1998

Purpose : The proliferative nuclear antigen Ki-67, present in all cell cycle phases except G0, is a useful marker for the detection of proliferative cells in vivo. MIB I has been found to recognize an antigen in formalin-fixed and wax-embedded material. The aim of this study was to assess the efficacy of MIB-1 expression as a marker of representing the status of mesangial cell proliferation in renal tissues. Methods : Immunohistochemical staining for Ki-67 Ag using monoclonal antibody MIB-1 (Immunotech,505) were performed in 41 renal tissuses which were obtained by percutaneous renal biopsy done between January 1994 and December 1996. Results : In both glomeruli and renal tubules, MIB-1 expression was observed only in 2 of 18 ($11.1\%$) cases of IgA nephropathy, in 2 of the 4 ($50\%$) cases of mebranoproliferative glomerulonephritis, in 4 of the 5 ($80\%$) cases of poststreptococcal glomerulonephritis. But MIB-1 expression was not detected in all cases of minimal lesion and membranous nephropathy. Renal tubules In another 7 cases of IgA nephropathy were MIB-1 positive. Conclusion : MIB-1 expression in renal tissues may relate to the cell proliferation in glomeruli and renal tubules. But the efficacy of MIB-1 expression as a marker of mesangial cell proliferation may reveal a limited value because of it's lower positive rate in IgA nephropathy.

• Journal of Chest Surgery
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• v.40 no.8
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• pp.529-535
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• 2007

Background: An imbalance between oxidants and antioxidants leads to oxidative stress, and this has been proposed to play an important role in the pathogenesis of lung neoplasm. Apurinic/apyrimidinic endonuclease-1/redox factor-1 (APE/ref-1) is a multifunctional protein involved in DNA base excision repair and the redox regulation of many transcription factors. However, the alteration of the expressed levels of APE/ref-1 in non-small cell lung cancer is unknown. Material and Method: Forty-nine patients with surgically resected non-small cell lung cancer (NSCLC) were included in this study. Immunohistochemical staining with APE/ref-1 antibodies was performed, and their expressions were analyzed via Western blotting for specific antibodies. Result: APE/ref-1 was localized at the nucleus and mainly in the non-tumor region of the NSCLC tissue specimens; it was expressed in the cytoplasm and nucleus of the NSCLC. The nuclear and cytoplasmic expressions of APE/ref-1 in lung cancers were markedly up-regulated in the NSCLC, and this was correlated with the clinical stage. Catalase, as first-line antioxidant defense, was dramatically decreased in the NSCLC. Conclusion: Taken together, our results suggest that APE/ref-1, and especially cytoplasmic APE/ref-1, was upregulated in the lung cancer regions, and this may contribute to the compensatory defense system against oxidative stress. A low expression of catalase might have fundamental effects on the extracellular redox state of lung tumors, along with the potential consequences for the tumors.

• Journal of Life Science
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• v.27 no.4
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• pp.383-389
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• 2017

This study intends to analyze the contents of rutin, procyanidin B3, quercetin, kaempferol, known to have antioxidant, anti-inflammatory and anti-carcinogenic effects, among the polyphenol type contained in the grape pruning stem extracts (GPSE), utilizing grape stems being discarded after harvest, measure the effects on the skin moisture, inhibition of skin cell proliferation, anti-inflammatory on the damaged skin of a HR-1 mice induced with UVB, and verify the applicability as a material for functional food and functional cosmetics. The results of verifying the photoprotection effects through the skin proliferation control through of GPSE showed similar result to suncream was achieved at the GPSE concentration of 2,000 mg/kg on the epidermis (p<0.05). The results showed anti-inflammatory effects on all groups applied with GPSE as compared to the control group irradiated with UVB, but at the GPSE concentration of 1,000 mg/kg, a lower COX-2 protein expression at 8%, lower than the 22% of suncream, was observed to achieve an excellent anti-inflammatory effect (p<0.05). The results of this study confirmed the existence of active polyphenol type, such as rutin, kaempferol, querocetin and procyanidin B3, within the GPSE, and GPSE has improvement effects on moisturizing effects, skin proliferation control effect, inflammatory control effect and improvement effects on the skin barrier function through UV ray damage. GPSE is a functional ingredient with a potential for skin protection effects, and has high utilization as an ingredient for functional food and functional cosmetics.

• Tuberculosis and Respiratory Diseases
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• v.40 no.6
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• pp.659-668
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• 1993

Background: p53 is currently considered as a tumor suppressive gene product, and its alterations are suggested to be involved in several human malignancies, including non-small cell lung cancers. p53 expression rates are variable in many reports and among cell types. Also, whether the phase of p53 expression is early or late during carcinogenesis is not certain. Thus, We have investigated to evaluate p53 expression rates of the various cell types and tissues and identify expression phase (early or late). Method: We obtained 71 tissue from 50 non-small cell lung cancer patients and performed the simple immunohistochemical staining using nonspecific monoclonal antibody(NCL-p53DO7). Results: 1) In non-small cell lung cancer patients. the expression rate of lungs(46.5%) is higher than that(25.0%) of lymph nodes. But, there is no significant difference between two groups. 2) Among the various cell types, p53 expression rates in squamous cell carcinoma and adenocarcinoma are 58.3% and 50.0% respectively without significant difference. 3) p53 expression rates in various stages are 33.3%, 60.0%, 40.0%, 60.0% and 66.7% in stage I, II, IIIa, IIIb and IV, respectively with no significant difference. 4) p53 expression rates in the various T parameters are 33.3%, 50.0%, 16.7% and 100% in T1, T2, T3 and T4, respectively and p53 expression rates in the various N parameters are 27.3%, 22.2% and 25.0% in N1, N2 and N3, respectively. There are no significant differences in the expression rates among varous T & N parameters. 5) p53 expression rates of lymph nodes in patients who have positive stains in lungs are 12.5% and 50.0% in N1 and N2. 6) p53 expression rates of all lymph nodes in patients who have negative stains in lungs are 0.0%. Conclusion: The above results show that p53 expression rate in non-small cell lung cancers is not correlated with cell type and progression of stage and it is thought to need further investigations about at what phase p53 expression influences the development and progression of lung cancers.