자궁경부암 조직에서 동시항암화학방사선치료의 반응 예측인자로서 p53, Bc1-2, Bax, Survivin의 분석: 예비보고

김현진 ( Hyun Jin Kim ),이정필 ( Jung Pil Lee ),박재선 ( Jae Sun Park ),임현이 ( Hyun Ee Lym ),유희석 ( Hee Sug Ryu ),장기홍 ( Ki Hong Chang ) 대한산부인과학회 2008 Obstetrics & Gynecology Science Vol.51 No.2

Objective: The objective of this study is to explore changes of the expression of apoptosis-regulating genes, p53, Bc1-2, Bax, and Survivin in cervical cancer tissues during concurrent chemoradiotherapy (CCRT). Methods: The expressions of apoptosis-regulating genes were evaluated in 14 patients with invasive squamous cell carcinoma of the cervix, who underwent 6 cycles of CCRT using weekly cisplatin, with immunohistochemical staining and Western blot of p53, Bc1-2, Bax, Survivin. Specimens of the cervical carcinomas were obtained by punch biopsy before starting CCRT and just after 2nd and 4th cycle of weekly cisplatin. We analysed the results with clinicopathological factors of the patients. Results: Among 14 patients, 2 (14.3%) patients did not respond to CCRT. In these 2 cases, p53 and Bax were not expressed before and during therapy, while the expressions of Bc1-2 and Survivin were increased. Complete response and partial response were obtained in 11 cases (78.6%) and 1 case (7.1%), respectively. In this reponse group, Bc1-2 expressiondecreased seriallyduring the treatment, but the expression ofother apoptosis-regulating genes showed no significant change. There was no correlation between the clinicopathological prognostic factors and expressions of p53, Bc1-2, Bax, and Survivin. Conclusion: In this preliminary study, we deduced that Bc1-2 expression might predict response of CCRT even before completing treatment. In order to establish this possibility, a large scaled study should be needed in the future.

악성 흑색종과 멜라닌 세포성 모반의 bc1 - 2 단백의 발현 양상

김현철,김윤규,채영수,서기석,김상태 ( Hyun Cheol Kim,Yun Kyew Kim,Young Soo Chae,Kee Suck Suh,Sang Tae Kim ) 대한피부과학회 1997 대한피부과학회지 Vol.35 No.2

Background; The bcl-2 is a newly known oncogene involved in tumorigenisis by blocking apoptosis or programmed cell death. Overexpression of bcl-2 protein has been detected in a variety of human malignancies. However, recent studies of the expression of bcl-2 protein in human melanoma and melanocytic nevus have been controversial. Objective : The purpose of this study was to examine whether there are any differences in the expression of bcl-2 protein between melanocytic nevus and rnalignant melanoma. Methods : Immunohistochemical analysis of bcl-2 protein expression was performed on the formalin-fixed, paraffin-embedded tissue sections of 22 melanocytic nevus and 29 malignant melanomas(20 primary and 9 metastatic) using anti bcl-2 monoclonal antibody with an avidin-biotin peroxidase complex procedure. Results : The results were as follows. 1. The positive rate for bcl-2 protein was observed in 95.4% (21/22) of melanocytic nevus and 95.0%(19/20) of primary malignant melanomas. Therefore, there was no significant difference between the two groups in the positive rate for bcl-2 prtoein. 2. The percentage of stained cells and the staining intensity of bcl-2 protein were significantly increased in melanocytic nevus compared to malignant melanoma(p<0.05). 3. The positive rate for bcl-2 expression of metastatic malignant melanoma[44.4% (4/9)] was significantly decreased compared to that of primary malignant melanoma[95.0%(19/20) ](p<0. 05). But, there was no significant difference betweeen tumor thickness and histological type of malignant, melanoma in the expression of bcl-2 protein. 4. In melanocytic nevus, immunoreactivity of bcl-2 protein gradually diminished or even disappeared towards the deep dermis Conclusion : the bcl-2 expression was decreased in malignant melanoma compared to melanocytic nevus. It. suggests that the loss of bcl-2 expression may play a significant role in the progression and metastasis of malignant melanoma. (Kor J Dermatol 1997;35(2): 284-291)

일측 요로폐쇄 마우스 모델에서 b치-2 및 Fas 유전자 발현에 미치는 Renin-Angiotensin System의 영향

송정헌 ( Jung Hun Song ),이철호 ( Chul Ho Lee ),이강욱 ( Kang Wook Lee ),신영태 ( Young Tai Shin ) 대한신장학회 2003 Kidney Research and Clinical Practice Vol.22 No.2

목적 : 요로 폐쇄에 의한 수신증은 장기적으로 지속될 경우 신장조직의 위축 및 섬유화로 인해 신부전증으로 진행하게 되는 중요한 원인 중의 하나이다. 이러한 조직손상 메카니즘에는 renin-angiotensin system (RAS)의 활성화와 신장세포들의 apoptosis 과정이 관여할 것으로 추정되고 있다. 연구자는 일측요로폐쇄 (unilateral ureteral obstruction, UUO) 마우스 모델에서 신조직내 RAS의 역할과 TGF-β 및 apoptosis와 관련 있는 bcl-2 및 Fas 유전자 발현 변화를 분석하고자 본 연구를 수행하였다. 방법 : CBA 마우스를 대상으로 수술적 방법으로 일측 요로를 결찰하였으며 72시간 후 희생시켜 신장조직내 유전자 발현을 RT-PCR 방법을 이용하여 측정하였다. 결과 : 일측요로폐쇄 수술을 받은 군 (UUO)에서는 sham 수술을 받은 군에 비하여 유의하게 혈압이 상승하고 신조직내 renin 유전자의 발현이 증가하여 본 모델에서 조기부터 RAS의 활성화를 확인할 수 있었다. TGF-β 유전자 발현은 비치료 UUO 군에 비하여 유의하게 낮았다. 신조직내 bcl-2 및 Fas 유전자 발현은 비치료 UUO 군에서 sham 군에 비하여 모두 유의하게 상승되어 있었으며 ACEI 치료 UUO 군에서는 bcl-2 유전자 발현이 비치료 UUO 군에 비하여 유의하게 감소하였다. 그러나 신조직내 Fas 유전자 발현은 비치료 UUO 군과 ACEI 치료군 사이에 유의한 차이를 보이지 않았다. 결론 : 이상에서 UUO에 의한 신조직 손상 모델에서 초기부터 bcl-2와 Fas 등 apoptosis 관련 유전자들의 발현 증가 및 RAS의 활성화가 신조직 손상기전에 중요한 요인 중의 하나로 작용할 것으로 생각된다. Background : The progression of hydronephrosis due to urinary obstruction is one of major causes of end stage renal failure. The activation of renin-angiotensin system (RAS) and renal cell apoptosis may be involved in this mechanisma. Methods : In order to investigate the role of RAS in renal injury mechanism and renal expression of TGF-β, bcl-2 and Fas in experimental unilateral ureteral obstruction (UUO) mouse model, 15 CBA strain mice were underwent sham operation (n=5), UUO without treatment (n=5) and UUO with angiotensin converting enzyme inhibitor (ACEI, enalapril, n=5) under ethyl ether anesthesia. Each group was sacrificed at 72 hours after surgery. Compertitive RT-PCR was performed for the estimation of renin, angiotensin II AT1 receptor(AT1 R), TGF-β, bcl-2, Fas and glyceraldehyde-3-phosphate-dehydrogenase (GAPDH) gene expression levels in the kidneys. Results : Systolic blood pressure (SBP) and renal gene expressions of renin and AT1 R in untreated control UUO group were significantly increased compared to sham operated group at 72 hours after surgery. The level of TGF-β and bcl-2 gene expressions of ACEI treated UUO group was significantly lower than that of untreated UUO group. On the other hand. Fas gene expression of ACEI treated UUO group was not significantly different from that of ACEI untreated UUO group. Conclusion : We speculate that the upregulation of bcl-2 and Fas gene expressions and the early activation of renin-angiotensin system of kidney are important factors of renal injury mechanism in this model.

Correlations of bcl-2 expression with clinicopathological features in breast cancer

Lee, Hy-De,Koo, Ja Yun,Jung, Woo Hee Yonsei University, College of Medicine 1997 Yonsei medical journal Vol.38 No.4

신경외배엽 기원의 피부 종양에서 bc1 - 2 단백의 면역조직화학적 연구

서재정,원영호,김성진,이승철,전인기 ( Jae Jeong Seo,Young Ho Won,Seong Jin Kim,Seung Chul Lee,Inn Ki Chun ) 대한피부과학회 1997 大韓皮膚科學會誌 Vol.35 No.4

Background: Protooncogene, bcl-2 is known to inhibit, apoptosis induced by various stimuli. Its expression has been reported in various fetal and adult tissues, and also in tumors of neural origin. Objective : The purpose of this study was to evaluate the expression of bcl-2 in a skin tumor of neuroectodermal origin and to estimate whether this expression was useful in the different,ial diagnosis of tumors of neural origin or not. Method : Immunohistochemical stains by the LSAB(labelled streptavidin biotin) method for bcl-2 protein were performed in normal special nerve end-organs and a skin tumor of neural origin. Results : The immunohistochemical findings revealed strong positive results in Meissners corpuscles, but weak week positive results in Vater-Pacini corpuscles. There were also strong positive results in neurilernmomas which were mostly composed of Schwann cells, but results were mostly negative in neurofibromas and neurofibrosarcomas which were composed primarily of endoneurial fibroblasts of mesodermal origin except a few cells of Schwann cell origin. Benign granular cell tumors arising from Schwann cells, and Merkel cell carcinoma known to arise from the Merkel cells of neural crest origin showed strong positive reactions. Conclusion : The strong expression of bcl-2 protein exclusively in the tumor of neuroectodermal origin suggests a useful indicator for the differential diagnosis of skin tumors of neural origin. (Kor J Dermatol 1997;35(4): 667-673)

태아 흰쥐의 창상치유에서 Fas, Fas Ligand 및 bcl-2의 발현

배태희,김한구,박대숭,김우섭,김승홍,이태진 대한성형외과학회 2004 Archives of Plastic Surgery Vol.31 No.2

Cutaneous wound healing in adult humans and higher vertebrate animals results in scar formation. In contrast, both human and animal fetuses at early gestational ages exhibit skin wound healing without scarring. A recent study has suggested that apoptosis occurs and plays an important role in achieving a decrease in cellularity during skin wound healing. The purpose of this study is to reveal the hypothesis the apoptosis may decreases the inflammatory infiltrates in fetal skin wound healing and may affect the fetal scarless wound healing.Open full-thickness incisional skin wounds were created on fetal rats at gestational ages 16 days(term= 21days). Wound were harvested at 24 hour(n=15), 72 hour(n=15), 120 hour(n=15). Adult skin wound was harvested at 24 hour(n=15), 72 hour(n=15), 120 hour(n =15). The wounds were fixed and stained with hematoxylin and eosin, TUNEL stain, immunohistochemical stain for Fas, Fas ligand, bcl-2. Fetal wounds was healed without scar formation and with regeneration of normal dermal and epidermal appendage architecture. Immunohistochemical staining for Fas, Fas ligand shows sparse positive cells in squamous epithelium of the both adult and fetus, there are no difference of expression between two groups. Immunohistochemical stainings for bcl-2 shows no positive cells in both adult and fetus. The apoptotic index of adult is 0.65±0.32 and fetus is 0.56 ±0.37(p=0.464), there is no significant difference stastically between two groups.These data indicate that apoptosis is not likely to be related to decreased infiltration of inflammatory cells that is main factor of scarless fetal wound healing.

탈진적 일회성 운동과 인위적 근손상에 의한 Caspase-3, Bcl-2, Bax mRNA 변화

최미리(Choi Mi-Lee),김재호(Kim Jae-Ho),김창균(Kim Chang-Gyun) 한국체육과학회 2006 한국체육과학회지 Vol.15 No.4

The purpose of this study was to investigate the caspase-3, Bcl-2, Bax mRNA and DNA fragmentation, for exercise and the artificial muscle injury induced the muscle injury. The mice(ICR) were housed two per cage in a temperature of 23~24℃ and controlled in an evironment with a cycle of 12:12h light-dark and provided with food and water. The exercise were canied out on an adapted treadmill in order for 2 animals to run at the same time. A total exercise time was 60min with the average speed of 24-25m/min. One section of Quadriceps muscle of the exercise group killed with cervical decapitation after physical activity for 1hour was cut from the RㆍL low limbs, frozen in liquid nitrogen and stored at -80℃ for analysis. The caspase-S, BcI-2, Bax transcript contents were estimated by RT-PCR DNA laddering was used to detect DNA fragmentation as an indicator of apoptosis. The results of the study can be summerized as follows ; First. caspase- 3 mRNA was significantly increased on rest, immediately after exercise and recovery at 1h, 5h, 12h after exercise in exercise group when compared with artificial muscle injury. Second, BcI-2 mRNA was significantly increased on rest, immediately after exercise and recovery at 1h, 5h, 12h after exercise in exercise group. Also, the artificial muscle injury group was increased when compared with rest, but not significantly. Third, Bax transcript was significantly increased in rest, immediately after exercise and recovery at 1h, 5h, 12h after exercise in exercise group when compared with artificial muscle injury group. And Bax mRNA of exercise group peaked at recovery 1h, The artificial muscle injury group was also increased when compared with rest, but not significantly. n rest, immediately after exercise and recovery at i1h, 5h after exercise in artificial muscle injury group when compared with the exercise group at appeared peak point on recovery 12h after the injury. In the present study, it was observed that apoptotic measurements differed in quadriceps muscle after the treatments(exhausted exercise vs artificial muscle injury). For example, it was demonstrated that exercise group commenced DNA fragmentation at 5h, increased BcI-2 and caspase-3 transcript levels, Bax m-RNA in mice quadriceps muscle had a significantly low status when compared with artificial muscle injury group. Thus, exercise was adapted for the skeletal muscle myocite which had a programmed cell death.

출생 후 제대혈관 폐쇄에서의 세포고사

황경국(Kyung Kuk Hwang),김효수(Hyo Soo Kim),서정욱(Jung Uk Seo),채인호(In Ho Chae),손대원(Dae Won Sohn),오병희(Byung Hee Oh),이명묵(Myoung Mook Lee),박영배(Young Bae Park),최윤식(Yun Shik Choi),서정돈(Jung Don Seo),이영우(Young Woo L 대한내과학회 1998 대한내과학회지 Vol.55 No.6

Background: The mechanism of the closure of umbilical vessels is known to be multifactorial. In order to verify that apoptosis is one of the possible closure mechanisms, we studied to identify apoptosis in umbilical vessels and evaluate its mechanism by studying apoptosis-related gene and the relationship between the pattern of apoptosis and gestational age (GA). Methods: Twelve umbilical cords of GA of 37-42 weeks were obtained immediatly (less than 10 min. for minimal ongoing external influences) after birth. The presence of apoptotic cells was demonstrated by electron microscopy (EM) and terminal deoxynucleotidyl transferasemediated dUTP nick end labeling (TUNEL). Immnohistochemical staining and Western blotting were used for the analysis of the proteins of apoptosis-related gene. Results: Apoptosis of the smooth muscle cells of media and intima of umbilical vessels was identified at EM, regardless of GA from 37 to 42 weeks. The proportion of TUNEL(+) cells was 80% in intima, 40% in media, 80% in connective tissue of umbilical cord. The expressions of bax-a, bcl-Xs/L were strong in intima, in media and connective tissue, but those of bcl-2 were detected in only in connective tissue, regardless of GA in immunohistochemistry. The ratios of expressions of bax- a to bc1-2, bcl-Xs to bcl-XL, did not change with increasing GA from 37 to 42 weeks in Western blot- ting. Conclusion: Apoptosis was identified in umbilical vessels. The closure of umbilical vessels can be explained by apoptosis where the ratios of bax-a/bcl-2, bcl-Xs/ bcl-XL play an important role. The fact that there were no differences in the extent of apoptosis and the expressions of bax-a/bcl-2, bcl-Xs/bcl-XL according to GA, suggests that apoptosis of umbilical vessels is more dependent on the external stimuli during delivery than GA.

Technology, chronology and the role of crucible steel as inferred from iron objects of the ancient site at Junnar, India

Park, J.S.,Shinde, V. Academic Press 2013 Journal of archaeological science Vol.40 No.11

Ordinary iron objects from an ancient habitation site at Junnar in India, dating to the 2nd BC to AD 2nd century, were examined for their microstructure, chemical composition and age. The objects were mostly made of high carbon steel with a homogeneous microstructure consisting of fine spherical particles of carbide in the ferrite background, free of non-metallic inclusions. Their carbon concentration ranged from 0.7% to over 1.6% with one exception at 0.2%. Some of them contained trace amounts of silicon, manganese and sulfur while one object retained cavities due to volume contraction during solidification reactions. These features indicate that the objects examined constitute an early example of Indian steel making in crucibles. Evidence was found that basic techniques needed for the success of crucible steel technology were mostly available at Junnar at the time. The radiocarbon measurement on carbon samples extracted from one of the iron objects placed its date between 176 BC and AD 20, in agreement with the radiocarbon dates of three charcoal samples from the same site. The forgoing results support that crucible steel was produced in India at a much earlier date than previously supposed, to serve as a material for specific needs arising in daily life.

Apoptosis Detected by in Situ DNA end-extension in Osteosarcomas - In relation to p53 and Bcl-2 expression -

Park, Yong-Koo,Yang, Moon-Ho,Park, Hye-Rim,Kim, Youn-Wha,Lee, Ju-Hie The Korean Musculoskeletal Tumor Society 1997 대한골관절종양학회지 Vol.3 No.2

Objective : The objective of this study was to compare expression of various proto-oncogenes and rates of apoptosis in osteosarcoma patients. Modulation of apoptosis may influence resistance to chemotherapy and therefore affect the outcome of cancer treatment. Osteosarcoma is one of the most fatal malignancies in young adolescents and investigation of the role of apoptotic cell death is warranted in relation to chemotherapy and tumor outcome. Design : The terminal deoxynucleotidyl transferase to exposed 3'-hydroxyl termini of DNA (TUNEL method) staining method has been applied for the in situ detection of DNA double strand breaks. Patients : Thirty-three osteosarcomas in various stages of differentiation from twenty-nine patients were investigated immunohistochemically for p53, Bcl-2 and TUNEL method for apoptosis. Results and conclusion; We have found that higher level of wild type p53 were correlated with enhanced expression of apoptosis. Increased apoptosis rates were found in cases of negative Bcl-2 expression. In the present study, we have concluded that a significant proportion of osteosarcoma, a tumor in which resistance to chemotherapy often occurs, express high levels of p53 and low levels of Bcl-2. Our data provide further evidence for cross-talk between Bcl-2 and p53 and suggests that these genes are important determinants of drug-induced apoptosis.