1.Changes of RAD50 and p53 Expression in Ultraviolet B-Irradiated Skin.
Seung Chul LEE ; Won Sik PYO ; Jeong Eun LEE ; Young Ho WON
Korean Journal of Dermatology 2000;38(2):213-220
BACKGROUND: Ultraviolet irradiation causes various changes in cells or tissues including DNA damage, which results in changes of cell cycling, mutation and cell death such as apoptosis or necrosis. p53 has been studied widely to be as a regulator gene to modulate cell cycling. Previous report shows that RAD50 is a gene to be associated with p53 to repair damaged DNA. OBJECTIVE AND METHOD: In this study, we evaluated changes of RAD50 and p53 expression by ultraviolet B (UVB) irradiation in rat skin in vivo and in human fibroblasts (hF) in vitro. RESULTS: In rat skin irradiated with 400mJ/cm2 UVB, RAD50 protein and mRNA expression were decreased by from early stage after irradiation, and they were restored to its normal level after 12 h. In hF irradiated with 20mJ/cm2 UVB, change of RAD50 expression by UVB irradiation was similar to that of rat skin. On the contrary, p53 protein expression was increased by UVB irradiation from 6 h and 3 h after UVB irradiation in rat skin and hF, respectively, but p53 mRNA expression was not changed by UVB irradiation. CONCLUSION: RAD50 and p53 expression is modulated differently in UVB- irradiated skin. Further studies are warrented to evaluate functional relationship between the genes in repairing damaged DNA.
Animals
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Apoptosis
;
Cell Death
;
DNA
;
DNA Damage
;
Fibroblasts
;
Genes, Regulator
;
Genes, vif
;
Humans
;
Necrosis
;
Rats
;
RNA, Messenger
;
Skin*
2.Induction of Apoptosis and Expression of Apoptosis-related Gene Products in Response to Radiation in Murine Tumors.
Jinsil SEONG ; Nancy R HUNTER ; Luka MILAS
Journal of the Korean Society for Therapeutic Radiology 1997;15(3):187-196
PURPOSE: To analyze the involvement of apoptosis regulatory genes p53, p21waf1/cip1, bax and bcl-2 in induction of apoptosis by radiation in murine tumors. MATERIALS AND METHODS: The radiation-sensitive ovarian carcinoma OCa-I, and the radiation-resistant hepatocarcinoma HCa-I were used. Tumors, 8 mm in diameter, were irradiated with 25 Gy and at various times after irradiation, ranging from 1 to 48 h, were analyzed histologically for apoptosis and by western blot for alterations in the expression of these genes. The p53 status of the tumors were determined by the polymerase chain reaction-single strand conformation polymorphism assay. RESULTS: Both tumors were positive for wild-type p53. Radiation induced apoptosis in OCa-I but not in HCa-I. Apoptosis developed rapidly, peaked at 2 h after irradiation and returned to almost the background level at 48 h. In OCa-I radiation upregulated the expression of p53, p21waf1/cip1, and the bcl-2/bax ratio was decreased. In HCa-I radiation increased the expression of both p53 and p21waf1/cip1, although the increase of the latter was small. The bcl-2/bax ratio was greatly increased. In general the observed changes occurred within a few hours after irradiation, and either preceded or coincided with development of apoptosis. CONCLUSIONS: The development of apoptosis required upregulation of both p53 and p21waf1/cip1 as well as a decrease in bcl-2/bax ratio. In contrast, an increase in bcl-2/bax ratio prevented apoptosis in the presence of upregulated p53 and p21waf1/cip1. These findings indentified the involvement of multiple oncogenes in apoptosis regulation in vivo and demonstrate the complexity that may be associated with the use of a single oncogene assessment for predicting the outcome of cancer therapy with cytotoxic agents.
Apoptosis*
;
Blotting, Western
;
Cytotoxins
;
Genes, Regulator
;
Oncogenes
;
Up-Regulation
4.Role of Synovial Fibroblasts in Rheumatoid Arthritis.
Hanyang Medical Reviews 2005;25(2):36-42
Rheumatoid arthritis (RA) is a chronic inflammatory disease characterized by synovial tissue proliferation with progressive joint destruction. The etiology of RA remains unknown, but many factors, including autoimmunity, cytokines and genetic factors, participate in its pathogenesis. There is growing evidence that activated fibroblast like synoviocytes (FLS), as part of a complex cellular network, play an important role in the pathogenesis of RA. It has been understood that proinflammatory cytokines secreted from macrophages and lymphocytes may influence the activation of FLS, but invasive and aggressive behaviour of RA-FLS maintained even in the absence of inflammatory stimuli. This kind of partial transformation is characterized by alterations in the expression of regulatory genes such as p53 and signaling cascade, as well as changes in pathway leading to apoptosis. Under the influences of proinflammatory cytokines in rheumatoid joints, RA-FLS is actively involved in the matrix degradation through the production of matrix metalloproteinases (MMP) and cathepsin. In addition, activated RA-FLS exert specific effects on other cell types such as macrophages and lymphocytes. While careful mapping of cytokine networks a decade ago led to the successful development of anti-cytokine therapy, the elucidation of gene mutations and detailed signaling transduction pathways that are specific to RA as well as mechanisms of action of MMP may provide the new targets for novel therapeutic interventions for RA.
Apoptosis
;
Arthritis, Rheumatoid*
;
Autoimmunity
;
Cathepsins
;
Cytokines
;
Fibroblasts*
;
Genes, Regulator
;
Joints
;
Lymphocytes
;
Macrophages
;
Matrix Metalloproteinases
5.Relationship between HPV Infection and bcl-2 Protein Expression and Apoptosis in Invasive and In Situ Squamous Cell Carcinoma of the Uterine Cervix.
Myoung Ja CHUNG ; Kyu Yun JANG ; Woo Sung MOON ; Myoung Jae KANG ; Dong Geun LEE
Korean Journal of Pathology 1999;33(9):702-708
Human papillomavirus (HPV) 16/18 is a causative agent of uterine cervical carcinoma. HPV 16/18 can alter cell cycle regulation through apoptosis. Bcl-2 is an important regulatory gene of apoptosis. A study was done to evaluate the relation between HPV 16/18 and bcl-2 and apoptosis in 21 cases of carcinoma in-situ (CIS), 5 cases of microinvasive carcinoma and 23 cases of invasive squamous cell carcinoma. HPV 16/18 was detected by hybrid capture system (HCS), bcl-2 protein by immunohistochemical method and apoptosis by using the hematoxylin-eosin stained slide. The results were as follows: Expression of the bcl-2 protein was 43% (9/21) in CIS and 26% (6/23) in invasive carcinoma. Expression of the bcl-2 protein was 42% (5/12) in CIS with HPV 16/18 infection, 44% in CIS without HPV 16/18 infection, 20% (2/10) in invasive carcinoma with HPV 16/18 infection and 31% (4/13) in invasive carcinoma without HPV 16/18 infection. Mean apoptotic index (mAI) was 3.36 in CIS, 5.23 in microinvasive and 6.25 in invasive carcinoma. mAI was 3.66 in CIS with HPV 16/18 infection, 2.86 in CIS without HPV 16/18 infection, 6.18 in invasive carcinoma with HPV 16/18 infection and 6.30 in invasive carcinoma without HPV 16/18 infection. Based on these results, we conclude that there are no correlation between HPV infection and bcl-2, and between HPV infection and apoptosis in invasive and in situ carcinoma of the uterine cervix, and apoptosis is increased according to tumor progression.
Apoptosis*
;
Carcinoma, Squamous Cell*
;
Cell Cycle
;
Cervix Uteri*
;
Female
;
Genes, Regulator
;
Humans
6.Cystic fibrosis of pancreas and nephrotic syndrome: a rare association.
Selvi KELEKCI ; Musemma KARABEL ; Aydin ECE ; Velat SEN ; Ali GUNES ; Ilyas YOLBAS ; Cahit SAHIN
Korean Journal of Pediatrics 2013;56(10):456-458
Cystic fibrosis (CF) is a genetic disease with autosomal recessive inheritance and is common in Caucasian people. The prevalence of this disease is between 1/2,000 and 1/3,500 live births, and the incidence varies between populations. Although the CF transmembrane conductance regulator gene is expressed in the kidneys, renal involvement is rare. With advances in the treatment of CF, life expectancy has increased, and some previously unobserved disease associations are now seen in patients with CF. It is important to follow patients with CF for possible abnormalities that may accompany CF. In this paper, we present two rare cases of CF accompanied by nephrotic syndrome.
Child
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Cystic Fibrosis*
;
Genes, Regulator
;
Humans
;
Heredity
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Incidence
;
Kidney
;
Life Expectancy
;
Nephrotic Syndrome*
;
Pancreas*
;
Prevalence
7.Establishment of stable subline of K562 cells overexpressing high mobility group B1 protein.
Fan-Zhi YAN ; Jin-Song YAN ; Jia ZHAO ; Wei-Ping LI ; Xue-Yu CHEN ; Yan YANG ; Shu-Mei RAO ; Jing JIN
Journal of Experimental Hematology 2011;19(1):1-5
This study was aimed to establish a stable subline of K562 cells (K562-HMGB1) overexpressing HMGB1 protein and K562-HMGB1 sublines served as control, so as to provide a basis for exploring the role of hmgb1 gene in occurrence and development of leukemia and their mechanism. Protein-coding gene of hmgb1 was amplified by PCR with cDNA as template, which was synthesized by reverse transcription from total RNA extracted from U937 cells. The PCR-amplified hmgb1 gene was ligated into PMD18-T vector (PMD18-T-HMGB1 vector), and then transformed into E. coli strain DH5α. DH5α containing PMD18-T-HMGB1 vector were grown on LB agar plate supplemented with 100 µg/ml ampicillin overnight. The single ampicillin-selected DH5α clone was picked for culturing overnight and then harvested for plasmid extraction. The extracted plasmid was characterized to contain hmgb1 gene digested with the desired restriction enzymes of KpnI/XhoI. The correctness of hmgb1 sequence was confirmed with DNA sequencing. The insert of hmgb1 gene contained in PMD18-T-HMGB1 vector was cut out with restriction enzymes of KpnI/XhoI and then ligated into eukaryotic expression vector pcDNA3.1 to form pcDNA3.1-HMGB1 vector. 10µg of pcDNA3.1-HMGB1 or pcDNA3.1 plasmid was separately electroporated into K562 cells. At 48 hours after electroporation the cells were cultured with G418 at a final concentration of 800 µg/ml for over 2 weeks. Finally stably transfected sublines of K562 cells containing hmgb1 gene (K562-HMGB1), and of K562 containing pcDNA3.1 vector (K562-pcDNA3.1) served as a control, were obtained. The transcriptional or translational expression of hmgb1 gene was detected with RT-PCR or Western blot, respectively, to testify transfected efficiency and validity of stable subline of K562-HMGB1. The results indicated that the eukaryotic expression vector pcDNA3.1-HMGB1 plasmid was successfully constructed and was electroporated into K562 cells. The transcriptional or translational expression of hmgb1 gene in the stable subline of K562 cells containing hmgb1 gene was overexpressed. It indicated that stable subline of K562-HMGB1 cells was successfully established. It is concluded that the stable sublines of K562-HMGB1 cells or K562-pcDNA3.1 cells are successfully established, which provides a basis for exploring the roles and mechanisms of hmgb1 gene in leukemogenesis and development of leukemia.
Gene Expression
;
Genes, Regulator
;
Genetic Vectors
;
HMGB1 Protein
;
genetics
;
Humans
;
K562 Cells
;
metabolism
;
Plasmids
;
Transformation, Genetic
8.Improvement of natamycin production in an industrial strain by heterologous expression of the afsRS(cla) global regulatory genes.
Zhengsheng TAO ; Yemin WANG ; Hualiang ZHENG ; Meifeng TAO
Chinese Journal of Biotechnology 2015;31(5):744-751
The afsRS(cla) global regulatory genes from Streptomyces clavuligerus activate the production of two antibiotics in Streptomyces lividans. In this study, we gained an increase of 38% in the production of natamycin (3.56 g/L) in an industrial strain Streptomyces gilvosporeus TZ1401 through the integration of pHL851 that bears the afsRS(cla) global regulatory genes into its genome. We discovered by quantitive real-time reverse transcription PCR (qRT-PCR) that the expression of 6 genes of the natamycin biosynthetic gene cluster were improved from 1.9 to 2.7 times. This suggests that afsRS(cla) improve the production of natamycin through increased transcription. This study provides a good example for applying afsRS(cla) in high yield breeding of industrial antibiotic producers.
Anti-Bacterial Agents
;
biosynthesis
;
Genes, Regulator
;
Industrial Microbiology
;
Multigene Family
;
Natamycin
;
biosynthesis
;
Streptomyces
;
genetics
9.Cystic Fibrosis: Case Report.
Si Hyun PARK ; Hyun ju LEE ; Ji Hye KIM ; Chol Heui PARK
Journal of the Korean Radiological Society 2002;47(6):693-696
Cystic fibrosis is a autosomal recessive genetic disease. Among Caucasians, it is the most common cause of pulmonary insufficiency during the first three decades of life. The prevalence of cystic fibrosis varies according to ethnic origin: it is common among Caucasians but rare among Asians. We report a case in which cystic fibrosis with bronchiectasis and hyperaeration was revealed by high-resolution CT, and mutation of the cystic fibrosis conductance transmembrane regulator gene (CFTR) by DNA analysis.
Asian Continental Ancestry Group
;
Bronchiectasis
;
Cystic Fibrosis*
;
DNA
;
Genes, Regulator
;
Humans
;
Prevalence
10.Estimation of the Genetic Substitution Rate of Hanwoo and Holstein Cattle Using Whole Genome Sequencing Data.
Genomics & Informatics 2018;16(1):14-20
Despite the importance of mutation rate, some difficulties exist in estimating it. Next-generation sequencing (NGS) data yields large numbers of single-nucleotide polymorphisms, which can make it feasible to estimate substitution rates. The genetic substitution rates of Hanwoo and Holstein cattle were estimated using NGS data. Our main findings was to calculate the gene's substitution rates. Through estimation of genetic substitution rates, we found: diving region of altered substitution density exists. This region may indicate a boundary between protected and unprotected genes. The protected region is mainly associated with the gene ontology terms of regulatory genes. The genes that distinguish Hanwoo from Holstein in terms of substitution rate predominantly have gene ontology terms related to blood and circulatory system. This might imply that Hanwoo and Holstein evolved with dissimilar mutation rates and processes after domestication. The difference in meat quality between Hanwoo and Holstein could originate from differential evolution of the genes related to these blood and circulatory system ontology terms.
Animals
;
Cattle*
;
Diving
;
Gene Ontology
;
Genes, Regulator
;
Genome*
;
Meat
;
Mutation Rate