2.The Translocation of Nuclaer Factor Kappa B on Corneal Epithelial Cells after Ultraviolet B Irrradiation.
Do Hyung LEE ; Kyu Dae CHOI ; Choun Ki JOO
Journal of the Korean Ophthalmological Society 2002;43(1):171-177
PURPOSE: To seek the role of nuclear factor kappa B (NF-kB) on the corneal epithelial cell death after ultraviolet (UV) irradiation. METHODS: Human corneal epithelial cells transfected by Simian Virus 40 were used in this study. UVB(312 nm) located at 10cm distance from bottom (0.6 mW/cm2 ) was irradiated for 10, 20, 30, and 40 seconds. To measure the cytotoxicity, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) method was used. Translocation of NF-KB was examined by immunocytochemistry with anti NF-K B p65 antibody and Electrophoretic Mobility Shift Assay (EMSA). To confirm the role of NF-KB , sulfasalazine, a specific inhibitor of NF-KB (0.5 mmole), was pretreated for 30 minutes before irradiatrion, and cytotoxicity and translocation of NF-KB was evaluated. RESULTS: UV irradiation resulted in a significant decrease in viability of cultured human corneal epithelial cells, especially after 20 second duration. When HCECs were irradiated with UVB, the translocation of N F -KB was observed in immunocytochemistry. These translocation was peaked 2 hours after UV irradiation in EMSA. In HCECs pretreated with sulfasalazine, either the cellular death or the translocation of NF-KB was blocked. CONCLUSION: UV irradiation can translocate NF-KB on the cultured human corneal epithelial cells. The cellular death after UV irradiation was blocked by sulfasalazine, a potent inhibitor of translocation of NF-KB. These findings suggest that NF-KB plays an important role in cellular death after UV irradiation.
Electrophoretic Mobility Shift Assay
;
Epithelial Cells*
;
Humans
;
Immunohistochemistry
;
NF-kappa B
;
Simian virus 40
;
Sulfasalazine
3.Well-Differentiated Papillary Mesothelioma of the Tunica Vaginalis: A Case Study and Review of the Literature.
Seyda ERDOGAN ; Arbil ACIKALIN ; Handan ZEREN ; Gulfiliz GONLUSEN ; Suzan ZORLUDEMIR ; Volkan IZOL
Korean Journal of Pathology 2014;48(3):225-228
Well-differentiated papillary mesothelioma is an uncommon tumor of the testes that usually presents as a hydrocele. Here, we present the case of one patient who did not have a history of asbestos exposure. The tumor was localized in the tunica vaginalis and was composed of three pedunculated masses macroscopically. Microscopically, branching papillary structures with focal coagulative necrosis were present. In addition to immunohistochemistry, simian virus 40 DNA was also tested by polymerase chain reaction. This report presents one case of this rare entity, its clinical and macroscopic features, and follow-up results.
Asbestos
;
DNA
;
Follow-Up Studies
;
Humans
;
Immunohistochemistry
;
Mesothelioma*
;
Necrosis
;
Polymerase Chain Reaction
;
Simian virus 40
;
Testis
5.The Role of Nuclear Factor Kappa B on Lens Epithelial Cells after Ultraviolet B Irradiation.
Do Hyung LEE ; Jung Kweon KIM ; Sae Hoon OH ; Eung Kwon KIM ; Choun Ki JOO
Journal of the Korean Ophthalmological Society 2004;45(3):500-506
PURPOSE: This study was performed to determine the role of nuclear factor kappa B (NF-kappa B) on the lens epithelial cell death after ultraviolet (UV) irradiation. METHODS: Simian virus 40 transfected human lens epithelial cells (HLE B-3 cells) were used in this study. UVB located at 10cm from the bottom was irradiated during 1, 2, 3 and 4 minutes. To measure the cytotoxicity MTT assay was used. Translocation of NF-kappa B was examined by immunocytochemistry with anti NF-kappa B p65 antibody and electrophoretic mobility shift assay (EMSA). To confirm the role of NF-kappa B, the cells were pretreated with sulfasalazine, a specific inhibitor of NF-kappa B, for 30 minutes before irradiation, and cytotoxicity and translocation of NF-kappa B were evaluated. RESULTS: UV irradiation produced a progressive cytotoxic effect in cultured HLE B-3 cells after 1 minute and maximum cytotoxicity was reached after 3 minutes irradiation. When HLE B-3 cells were irradiated with UVB, the translocation of NF-kappa B was observed in immunocytochemistry. These translocations were peaked 6 hours after UV irradiation in EMSA. In HLE B-3 cells pretreated with sulfasalazine, the translocation of NF-kappa B was blocked. The cellular death after UV irradiation was markedly blocked by sulfasalazine. UV irradiation can translocate NF-kappa B and sulfasalazine is a useful blocking agent in this pathway. In addition, sulfasalazine can prevent cellular death after UV irradiation. CONCLUSIONS: These findings suggest that NF-kappa B plays an important role in cellular death after UV irradiation.
Electrophoretic Mobility Shift Assay
;
Epithelial Cells*
;
Humans
;
Immunohistochemistry
;
NF-kappa B*
;
Simian virus 40
;
Sulfasalazine
;
Transcription Factor RelA
6.Immunohistochemical study of p21 and p53 expression in ameloblastoma.
Dong Joon SHIN ; Hoon MYOUNG ; Kyeng Kyun HWANG ; Myung Jin KIM
Journal of the Korean Association of Oral and Maxillofacial Surgeons 2003;29(4):199-205
The p53 protein was discovered in 1979 as cellular 53-kD nuclear phosphoprotein bound to the large transforming antigen of SV40 virus. P21WAF1/CIP1, which has been described as the critical downstream mediator of p53, is known to suppress DNA replication and arrest the G1 cell cycle by quaternary complex with cyclin D, cyclin-dependent kinase(CDK) and proliferating cell nuclear antigen(PCNA). In these days, some studies shows that the p21 can be induced by independent pathways. There are various reports about the expression of p21 (67%.82.4%) in oral squamous cell carcinoma. But these studies are mostly done in malignant tumor not in benign tumor. So we decided to study the expression of p21 in ameloblastoma and the relationship between p53 and p21 as a downstream mediator of p53 in ameloblastoma. We investigated the expression of p21 and p53 with the method of immunohistochemistry. We selected 30 cases of ameloblastoma tissue blocks (acanthomatous type: 5 cases, follicular type: 8 cases, plexiform type: 17 cases) imbedded in paraffin. We used 30 cases of normal gingival tissues and 30 cases of squamous cell carcinoma tissues (SCC) respectively and compared their results with those of ameloblastoma. We made slides with the streptavidin-biotin methods and used monoclonal antibody DO-7 (Novocastra, Newcastle, United Kingdom) as p53 antibody and monoclonal antibody M7202 (DAKO, California, U.S.A.) as p21 antibody. We used Pearson's correlation coefficient to analyse the relationship. The results were as follows: 1. p21 was expressed in ameloblastoma about 30% and this is lower than that of normal gingiva and SCC. 2. In normal gingiva and ameloblastoma, p21 expression was correlated with p53 expression. 3. In SCC, p21 were expressed about 83.3% and this is more than that of p53. But there was no correlation between p21 and p53 expression. We confirmed p21 expression and relation with p53 in ameloblastoma. But, to confirm the function of p21, more studies about p21 expression in malignant ameloblastoma and ameloblastic carcinoma are needed.
Ameloblastoma*
;
Ameloblasts
;
California
;
Carcinoma, Squamous Cell
;
Cell Cycle
;
Cyclin D
;
DNA Replication
;
Gingiva
;
Immunohistochemistry
;
Paraffin
;
Simian virus 40
7.Construction and reversion of pCTGTKlox for reversible hepatocyte immortalization.
Yao-kai CHEN ; Jun-gang LI ; Yu-ming WANG
Chinese Journal of Hepatology 2006;14(3):222-223
Cell Line
;
Clone Cells
;
cytology
;
metabolism
;
Cloning, Molecular
;
Genetic Vectors
;
genetics
;
Hepatocytes
;
cytology
;
metabolism
;
Humans
;
Simian virus 40
;
genetics
;
Transfection
8.Immortalization of human precartilaginous stem cells by transfecting SV40Tag.
Junfang, WANG ; Huang, FANG ; Renyun, XIA ; Anming, CHEN ; Hao, CHENG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2009;29(2):231-4
Immortalized human precartilaginous stem cells (IPSCs) were established to provide stable cell resource for the study of the molecular mechanism of gene targeting on the differentiation of PSCs. Plasmid pCMVSV40T/PUR containing simian virus 40 large T antigen gene (SV40Tag) was transfected into human PSCs by using lipofectin transfection. Colonies were isolated by puromycin selection and expanded by multiple passages. Immunohistochemistry, RT-PCR and Southern blotting were used to identify the transfected cells and to detect the expression and integration of SV40Tag in expanded cell lines. The positive colonies were isolated and subcultured, designated immortalized precartilaginous stem cells (IPSCs), which were confirmed as fibroblast growth factor receptor-3 (FGFR-3) positive cells by immunohistochemistry and RT-PCR. SV40Tag cDNA was found in cultured IPSCs of passage 8 by Southern blotting, and the expressions of SV40Tag mRNA and protein were confirmed by RT-PCR. These findings suggested that IPSCs strain with SV40Tag was constructed successfully.
Cartilage/*cytology
;
Cell Proliferation
;
Cell Transformation, Viral
;
Cells, Cultured
;
Fetus
;
Simian virus 40/*genetics
;
Stem Cells/*cytology
;
Transfection
9.No Detection of Simian Virus 40 in Malignant Mesothelioma in Korea.
Minseob EOM ; Jamshid ABDUL-GHAFAR ; Sun Mi PARK ; Joung Ho HAN ; Soon Won HONG ; Kun Young KWON ; Eun Suk KO ; Lucia KIM ; Wan Seop KIM ; Seung Yeon HA ; Kyo Young LEE ; Chang Hun LEE ; Hye Kyoung YOON ; Yoo Duk CHOI ; Myoung Ja CHUNG ; Soon Hee JUNG
Korean Journal of Pathology 2013;47(2):124-129
BACKGROUND: Simian virus 40 (SV40), a polyomavirus, was discovered as a contaminant of a human polio vaccine in the 1960s. It is known that malignant mesothelioma (MM) is associated with SV40, and that the virus works as a cofactor to the carcinogenetic effects of asbestos. However, the reports about the correlation between SV40 and MM have not been consistent. The purpose of this study is to identify SV40 in MM tissue in Korea through detection of SV40 protein and DNA. METHODS: We analyzed 62 cases of available paraffin-blocks enrolled through the Korean Malignant Mesothelioma Surveillance System and performed immunohistochemistry for SV40 protein and real-time polymerase chain reaction (PCR) for SV40 DNA. RESULTS: Of 62 total cases, 40 had disease involving the pleura (64.5%), and 29 (46.8%) were found to be of the epithelioid subtype. Immunostaining demonstrated that all examined tissues were negative for SV40 protein. Sufficient DNA was extracted for real-time PCR analysis from 36 cases. Quantitative PCR of these samples showed no increase in SV40 transcript compared to the negative controls. CONCLUSIONS: SV40 is not associated with the development of MM in Korea.
Asbestos
;
DNA
;
Humans
;
Immunohistochemistry
;
Korea
;
Mesothelioma
;
Pleura
;
Poliomyelitis
;
Polymerase Chain Reaction
;
Polyomavirus
;
Real-Time Polymerase Chain Reaction
;
Simian virus 40
;
Viruses
10.Detection of SV40 Large T Antigen in Malignant Lymphomas.
Young A KIM ; MeeSoo CHANG ; Jinho PAIK ; Sun Och YOON ; Yoon Kyung JEON ; Chul Woo KIM ; Ji Eun KIM
Korean Journal of Pathology 2009;43(4):312-316
BACKGROUND: The association of simian virus 40 (SV40) with certain types of human cancers, including malignant lymphomas, has been a topic of interest for some time. Although the virus is distributed worldwide, its incidences vary according to the specific types of tumors, and the epidemiological areas. The aim of this study was to investigate the frequency of SV40 in malignant lymphomas among Korean patients. METHODS: One hundred seventy three cases of malignant lymphomas were evaluated by immunohistochemical staining for SV40 large T antigen (TAg), using an extremely sensitive, tyramide based, catalyzed signal amplification method. RESULTS: From 158 non-Hodgkin's lymphomas, including 115 diffuse large B-cell lymphomas, and 15 Hodgkin's lymphomas, none of the cases were positive for SV40 TAg. CONCLUSIONS: SV40 does not appear to be related to the pathogenesis of malignant lymphomas among Koreans.
Antigens, Polyomavirus Transforming
;
Antigens, Viral, Tumor
;
Hodgkin Disease
;
Humans
;
Incidence
;
Korea
;
Lymphoma
;
Lymphoma, B-Cell
;
Lymphoma, Non-Hodgkin
;
Simian virus 40
;
Viruses