항암제 처리한 백혈병 세포주에서의 Apoptosis 발현

김삼용,윤소현,김현수,김종숙,윤환중,김진경,조덕연 충남대학교 의과대학 지역사회의학연구소 1998 충남의대잡지 Vol.25 No.1

The bcl-2 proto-oncogene encodes a 26 kD protein that promotes cell survival by blocking apoptosis. The bax protein is a member of the bcl-2 familly, now known to form heterodimers with the bcl-2 protein. The ratio of bax to bcl-2 is be critical in determining the fate of the cell in response to stimuli that can induce apoptosis. Extract of Pulsatilla Koreana (SB-31) showed promising antitumor activity in vitro with Topo I inhibitory action. In the present study, the relationship between apoptosis and the apoptosis related proteins, bcl-2 and bax were investigated in human leukemic cell lines HL-60, U-937 and CEM-CM3. All anticancer drugs(adriamycin, etoposide, camptothecin, SB-31) induced extensive apoptosis in HL-60, U-937 cells and CEM-CM3 cells. The expression of bcl-2 and bax protein were determined in cell lines by western blotting before and after incubation with anticancer drugs at different time points. 1) In HL-60 or U-937 cell lines, down regulation of bcl-2 and up-regulation of bax were found after incubation with ADR, VP-16 or camptothecin. 2) In HL-60 or U-937 cell lines, no significant change in bcl-2 or bax protein expression resulted after incubation with SB-31. 3) In CEM-CM3 cells, virtually no change was noted in bcl-2/bax expression after incubation with ADR, VP-16, camptothecin or SB-31. It is suggested that different leukemic cell lines use different pathways of apoptosis activation and a given cell may utilize different pathways of apoptosis activation in response to different cytotoxic agents.

폐경기 증후군 환자에서 Cyclofenil의 치료 효과

김덕윤,고은미,우정택,양인명,김진우,김영설,김광원,최영길 최신의학사 1992 最新醫學 Vol.35 No.10

항암제 처리한 백혈병 세포주에서의 Apoptosis 발현

김삼용,윤소현,김현수,김종숙,윤환중,김진경,조덕연 충남대학교 암연구소 1998 癌共同硏究所 硏究誌 Vol.2 No.1

The bcl-2 proto-oncogene encodes a 26 kD protein that promotes cell survival by blocking apoptosis. The bax protein is a member of the bcl-2 familly, now known to form heterodimers with the bcl-2 protein. The ratio of bax to bcl-2 is be critical in determining the fate of the cell in response to stimuli that can induce apoptosis. Extract of Pulsatilla Koreana (SB-31) showed promising antitumor activity in vitro with Topo I inhibitory action. In the present study, the relationship between apoptosis and the apoptosis related proteins, bcl-2 and bax were investigated in human leukemic cell lines HL-60, U-937 and CEM-CM3. All anticancer drugs(adriamycin, etoposide, camptothecin, SB-31) induced extensive apoptosis in HL-60, U-937 cells and CEM-CM3 cells. The expression of bcl-2 and bax protein were determined in cell lines by western blotting before and after incubation with anticancer drugs at different time points. 1) In HL-60 or U-937 cell lines, down regulation of bcl-2 and up-regulation of bax were found after incubation with ADR, VP-16 or camptothecin. 2) In HL-60 or U-937 cell lines, no significant change in bcl-2 or bax protein expression resulted after incubation with SB-31. 3) In CEM-CM3 cells, virtually no change was noted in bcl-2/bax expression after incubation with ADR, VP-16, camptothecin or SB-31. It is suggested that different leukemic cell lines use different pathways of apoptosis activation and a given cell may utilize different pathways of apoptosis activation in response to different cytotoxic agents.

정상과 갑상선 종양조직에서 사람 IGF-I 유전자의 발현

김성운,장현하,박상미,김덕윤,우정택,양인명,김진우,김영설,김광원,고석환,홍성화,최영길 경희대학교 유전공학연구소 1993 遺傳工學論文集 Vol.5 No.-

Many of the growth-promoting properties of growth hormone(GH) are mediated by insulin-like growth factor-I(IGF-I), a highly conserved circulating 70-amino acid peptide. Recent studies have shown that multiple mechanisms influence IGF-I gene expression, including transcription from two promoters, alternative RNA splicing, and variable polyadenylation. In thyroid tissue, thyroid stimulating hormone(TSH) and IGF-I are the most possible candidates for follicular cell proliferation and hypertrophy. Actually IGF-I had autocrine and paracrine effect for tissue growing. We prepared thyroid tumor tissue mRNAs using single step method for detecting IGF-I levels according to different tissues, i.e., thyroid adenoma or papillary thyroid carcinoma. We used Northern blot analysis for IGF-I mRNA and RNase protection assay (RPA) for IGF-I transcription start sites. For Northern blot, we used whole human IGF-I cDNA as a DNA probe and for RPA, we used IGF-I exon 1 containing noncoding promoter 1 as a riboprobe. We got good RNA bands from Northern blot analysis around 1 kb (IGF-IA) and 7.5 kb (IGF-IB) region. To clarify the amount of both IGF-IA and IB mRNAs, we measured autoradiographied signal of IGF-I mRNAs bands using densitometer. In IGF-IA signals, there's no change among liver and thyroid tissues, but in case of IGF-IB mRNA bands, the signal was markedly increased in thyroid carcinoma tissues than that of normal thyroid tissue (85% vs 14%). In the study of RPA, all thyroid tissues used the same transcription start sites as those of liver's. We concluded that that this different regulation of IGF-I mRNA was originated from tissue specificity. That meant some tissue specific transcription factor/s were related to tissue IGF-I expression.

한국인 당뇨병 및 비당뇨병 환자에서의 뇌혈관 질환 유무에 따른 PAI-1 촉진자 유전자형과 인슐린저항성에 관한 연구

오승준,김영설,박철영,김덕윤,김성운,양인명,김진우,최영길,팽정령,정경천 대한비만학회 2000 The Korean journal of obesity Vol.9 No.2

연구배경 : 혈전현상을 특징으로 하는 질환에서는 Plasminogen activator inhibitor-1 (PAI-1) 이 높은 활성도를 보이는데, PAI-1 치는 당뇨병, 심근경색증, 비만 등에서 높다고 밝혀진 바 있다. 또한 당뇨병 환자들의 합병증의 주요한 병인은 죽상경화증으로 혈전현상이 특징인 질환에서 증가하는 PAI-1이 당뇨병 환자에서 높다. 목적 : 정상인에서의 PAI-1 유전자 촉진자의 유전자형의 분포 및 혈액농도를 관찰하고, 당뇨병 및 뇌혈관 질환 환자군에서의 PAI-1 유전자 촉진자 유전자형의 분포 및 혈액농도를 측정하여 정상인과 차이점을 알아본다. 당뇨병 환자군에서의 혈장 PAI-1 치와 인슐린 저항성, 전구 인슐린 등과의 상관관계를 살펴보고, 인슐린저항성과 대혈관질환의 지표로 사용될 수 있는지 알아보았다. 방법 : 대상으로는 정상인 76명, 제2형 당뇨병 환자 56명, 뇌혈관질환이 동반된 제2형 당뇨병 환자 48명, 뇌혈관질환 환자 51명을 선택하여, 환자의 혈액에서 인슐린, 공복시 혈당, 전구인슐린, 중성지방, 총콜레스테롤 및 기타 생화학 검사 및 이학적 검사를 시행하였다. 환자의 DNA를 채취하여 전사개시 -675bp를 포함하는 대립형질 특이 시발체를 사용하여 중합효소 연쇄 반응을 실시하여, 그 유전자형을 판독하였다. 결과 : 정상 대조군 76명 (46.4±11.1세), 2형 당뇨병 환자 56명 (58.3±12.6세), 뇌경색증 환자 51명 (63.1±13.2세) 대상으로 하였다. PAI-1 촉진자 유전자형의 (4G/4G, 4G/5G, 5G/5G)빈도는 정상 대조군이 각각 23.7%, 75.0%, 1.3%, 뇌경색 환자군이 19.6%, 66.7%, 13.7%, 뇌경색이 동반된 당뇨병 환자군이 33.3%, 58.3%, 8.3% 였다. (X2=12.6, p=0.05). 이러한 사실은 서구인에 비해 4G/4G, 5G/5G 동형 유전자형이 낮은 결과였다. 각 군별 혈장 PAI-1 농도는 정상 대조군 13.4, 1.8 ~ 65.2 ng/mL (중앙값 , 범위 ) 2형 당뇨병 환자군 14.4, 2.9 ~ 47.8 ng/mL, 뇌경색 환자군 21.9, 6.2 ~ 154.7 ng/mL , 뇌경색이 동반된 2형 당뇨병 환자군 28.8, 3.2 ~ 139.3 ng/mL 로 차이를 보였다 (p=0.000). 전체 대상에서 PAI-1 촉진자 부위의 유전자형에 따른 PAI-1 활성도와 항원 농도는 차이를 보이지 않았다. 그러나 PAI-1 활성도는 혈중 중성지방, 전구인슐린, 체질량지수와 독립적인 상관관계를 보였다 (p=0.000, p=0.000 and p=0.005). 결론 : 결론적으로 PAI-1 촉진자 부위의 유전자형은 뇌경색증의 지표는 아니며, PAI-1 활성도를 결정짓는 인자는 유전적 요인보다는 혈중 중성지방, 전구 인슐린, 체질량지수와 같은 대사적 요인으로 생각된다. Plasminogen activator inhibitor-1 (PAI-1) is known be related to insulin resistance and several components of the large vascular disease. Notably, the high frequencies of diseases such as coronary heart disease or stroke are related to type 2 diabetes complications. We studied to find out whether the PAI-1 promother genotype could be a marker for cerebral infarction in type 2 patients. Subject patients were; 56 type 2 diabetics (age 58.3±12.6), 51 patients with cerebral infarction (age 63.1±13.2), 48 type 2 diabetics with cerebral infarction (age 64.8±9.3) , and 76 healthy control (age 46.4±11.1). The 4G/5G genotype of PAI-1 promoter was evaluated by polymerase chain reaction and endonuclease digestion. PAI-1 promoter genotype frequency (4G/4G, 4G/5G, 5G/5G) was 23.7%, 75.0% and 1.3% in healthy control, 17.9%, 67.9% and 14.3% in type 2 diabetes patients, 19.6%, 66.7% and 13.7% in cerebral infarction patients, 33.3%, 58.3% and 8.3% in type 2 diabetics with cerebral infarction (X^2=12.6, p=0.05). This finding is lower in frequency of 5G/5G homozygote than that reported in Caucasians. The plasma PAI-1 concentrations according to the disease were 13.4, 1.8 ~ 65.2 ng/mL (median, range) for healthy control, 14.4, 2.9 ~ 47.8 ng/mL for type 2 diabetes, 21.9 6.2 ~ 154.7 ng/mL for cerebral infarction , and 28.8, 3.2 ~ 139.3 ng/mL, for cerebral infarction with type 2 diabetes (p=0.000). In the all subjects, PAI-1 concentration and activity of PAI-1 promoter genotype did not show any significant difference. However, the PAI-1 activity was independently associated with serum triglyceride level, plasma proinsulin and BMI (p=0.000, p=0.000 and p=0.005 respectively). We concluded that PAI-1 genotype is not a marker for the cerebral infarction ; however, the genotype is related to PAI-1 concentration , and therefore it seems to be that metabolic factors such as triglyceride level or plasma proinsulin or BMI are more in relations with determining the PAI-1 concentration than the genotype.

심한 골병변으로 발현된 기능성 낭종성 부갑상선 선종

전숙,김영희,박지영,고관표,박철영,김덕윤,우정택,김성운,김진우,김영설,고석환 대한내분비학회 2003 Endocrinology and metabolism Vol.18 No.2

낭종성 부갑상선 선종과 심한 골병변을 동반한 부갑상선 기능항진증은 매우 드문 질환으로서, 저자들은 양측 고관절의 통증을 초기 주소로 내원한 환자에서 고칼슘혈증과 부갑상선 호르몬 증가, 골병변의 방사선적 소견을 통해 부갑상선 기능항진증을 진단하고, 경부 초음파와 컴퓨터 단층 촬영, 부갑상선 스캔검사 및 수술중 부갑상선 낭종액 검사 등을 통해 기능성 부갑상선 낭종의 한 종류인 낭종성 부갑상선 선종을 진단하고 수술적 제거를 통하여 정상화된 1예를 경험하였다. A cystic parathyroid adenoma is rare. A case of primary hyperparathyroidism, with the cystic formation of a parathyroid adenoma and a severe bony lesion, is reported. A 52-year-old male was admitted due to pain in both hips and for evaluation of hypercalcemia. The plasma level of the intact parathyroid hormone (iPTH) was elevated to 1424 pg/mL. Ultrasonography and the computed tomography revealed a parathyroid cyst on the left thyroid lower pole. Parathyroid scintigraphy detected a parathyroid adenoma. A radiograph showed a subperiosteal bone resorption on the phalanges, and a brown tumor (osteitis fibrosa cystica) on the femur shaft was noted. A surgical excision of the parathyroid adenoma was performed. The PTH level in the cystic fluid was increased. A histological examination confirmed a cystic parathyroid adenoma. The PTH level was normalized after the operation (J Kor SOC Endocrinol 18:214-220, 2003).

그람양성구균에 대한 Teicoplanin과 Vancomycin의 시험관내 항균력

최태열,김경숙,전용관,서일혜,김정욱,이웅수,안정열,김홍석,정재용,최효선,김덕언,유진우 대한감염학회 1994 감염 Vol.26 No.1

An increasing frequency of methicillin resistant S. aureus(MRSA), methicillin resistant coagulase negative staphylococci(MRCNS) and Enterococcal infection have been observed in recent years. Teicoplanin is a new glycopeptide antibiotic obstained from the Actinoplanes teicomycetius. The molecular structure and spectrum of antimicrobial activity of teicoplanin is simillar to those of vancomycin, and has been reported to have an excellent in vitro and in vivo effect against various gram-positive infections. Therefore, we evaluated the in vitor susceptibility of gram positive cocci, such as, S. aureus, coagulase negative Staphylococci(CNS), and Enterococci to teicoplanin and vancomycin. The total 253 strains consisted of MSSA(40), MRSA(53), MSCNS(47), MRCNS(48), and Enterococci(65). They were assayed by disc diffusion and agar dilution. During the study, 57% of S. aureus and 49% of CNS showed resistance to methicillin. The inhibitory diameter of teicoplanin was 15-20mm in MSSA, 12-19mm in MRSA, 13-24mm in MSCNS, 11-23mm in MRCNS, and 15-22mm in Enterococci respectively, and showed sensitivity in all but 8 strains(3.2%). The range of the minimum inhibitory concentration (MIC) of teicoplanin to MSSA, MRSA, MSCNS, MRCNS and Enterococci were 9.12-2.0㎍/ml, 0.25-2.0㎍/ml, & 0.25-32㎍/ml, 0.12-1.0㎍/ml respectively. One case of S. haemolyticus was resistant to teicoplanin (32㎍/ml) by the agar dilution method. Eight minor (3.2%) and one major(0.4%) error was observed when the MIC and disk diffusion data were correlated with teicoplanin. As for vancomycin the inhibitory diameter was 17-21mm in MSSA, 15-21mm in MRSA, 18-26mm in MSCNS, 18-25mm in MRCNS, and 16-22mm in Enterococci respectively. The range of the MIC of vancomycin to MSSA, MRSA, MSCNS, MRCNS, and Enterococci were 0.25-1.0㎍/ml, 0.25-4.0㎍/ml, 0.5-2.0㎍/ml and 0.5-2.0㎍/ml respectively. One minor error (0.4%) was seen with the vancomycin disk. The MIC90 of MSSA and MRSA exhibited the same results in teicoplanin (1.0㎍/ml, 1.0㎍/ml), and vancomycin(2.0㎍/ml, 2.0㎍/ml). MSCNS and MRCNS exhibited greater MIC90 with teicoplanin(4.0㎍/ml, 8.0㎍/ml) than vancomycin(2.0㎍/ml, 2.0㎍/ml). Incontrase Enterococci were more susceptible to teicoplanin(0.5㎍/ml) than to vancomucin (2.0㎍/ml). Results from this analysis indicated that both teicoplanin and vancomycin were very excellent for gram positive infections, especially those resistant to methicillin.

제2형 당뇨병 남성 환자에서 유리 테스토스테론과 성호르몬 결합 글로블린 농도

남기덕,김영설,박철영,오승준,김덕윤,우정택,김성운,양인명,김진우,최영길 대한당뇨병학회 2002 Diabetes and Metabolism Journal Vol.24 No.6

연구배경:인슐린저항성은 제2형 당뇨병과 심혈관질환의 주요한 위험 인자로 성호르몬과 상호 관계가 있다고 알려져 있다. 그러나 여성과는 달리 남성에서는 연령과 인슐린저항성에 따른 유리 테스토스테론과 성호르몬 결합 글로블린 농도의 변화에 대한 연구가 부족한 실정이다. 그러므로, 본 저자등은 제2형 당뇨병 남성 환자에서 정산인과 비교해서 유리 테스토스테론과 성호르몬 결합 글로블린 농도를 측정하고 연령에 따른 변화 정도를 알아보고자 하였다. 방법:대상 환자 모두에서 연령과 체질량지수, 총 콜레스테롤, 중성지방, 공복혈당과 인슐린 농도를 측정하였다. 혈중 유리 테스토스테론 농도는 방사면역측정법(radioimmunoassay)을 이용해서 측정하였고, 혈중 성호르몬 결합글로불린은 면역방사계측측정법(immunoradiometric assay)을 이용해서 측정하였다(Diagnostic System Laoratories, Wbster, TX, USA). 결과:1)제2형 당뇨병 남성 환자에서 정상 대조군 남성에 비해 성호르몬 결합글로불린은 104.1±35.0 vs 25.7±3.5 mole×10??로 의미 있게 높았으나(p<0.001), 유리 테스토스테론은 13.7±9.5 vs 13.6±6.5 ng/dL로 차이가 없었다. 2)연령과 성호르몬 결합 글로블린 사이의 상관 계수는 제2형 당뇨병 남성 환자에서 0.40로 중등고의 양의 상관 관계를 보였고(p<0.001), 정상 대조군 남성에서 0.11로 유의한 상관 관계를 보이지 않았다. 연령과 유리 테스토스테론 사이의 상관 계수는 제2형 당뇨병 남성 환자에서 0.08, 정상 대조군 남성에서-0.17로 모두에서 유의한 상관 관계를 보이지 않았다. 3)연령과 체질량지수를 보정한 후에 제2형 당뇨병 남성 환자와 정상 대조군 남성에서 혈중 인슐린 농도, 유리 테스토스테론과 성호르몬 결합 글로블린 사이에는 상관 관계가 없었다. 결론:제2형 당뇨병 남성 환자에서 정상 남성에 비해 성호르몬 결합 글로블린 농도가 증가되어 있었으며, 유리 테스토스테론은 차이가 없었다. 연령이 증가함에 따라 성호르몬 결합 글로블린이 제2형 당뇨병 남성 환자에서 정상 남성에 비해 증가 폭이 의미있게 컸으며, 유리 테스토스테론은 변화가 없었다. Background: Insulin resistance is a risk factor for cardiovascular disease and type 2 diabetes mellitus. There are many previous studies indicating that insulin lowers serum sex hormone-binding globulin levels, and there is inverse correlation between insulin resistance and serum sex hormone-binding globulin levels in women. However, in men, a limited number of studies are available to explain the effect of sex hormone on age and insulin. Therefore, the present study was undertaken to investigate the relationship among free testosterone, hormone-binding globulin and age in type 2 diabetic men and control subjects. Method: Age, body mass index, total cholesterol, triglyceride, fasting blood sugar, and insulin concentrations were examined on 89 type 2 diabetic men and 47 control subjects. The free testosterone level was measured by commercially available double-antibody system (Radioimmunoassay). The sex hormone-binding globulin level was also measured by commercially available double-antibody system(Immunoradiometric assay). Results: 1) Sex hormone-binding globulin level was significantly increased in patients with type 2 diabetes. However, there was no significantly difference in free testosterone level between the two groups. 2) Sex hormone-binding globulin was positively correlated with age (r=0.4, p <0.001) in patients with type 2 diabetes. Sex hormone-binding globulin and free testosterone were not correlated with age in control sujects. 3) Free testosterone and sex hormone-binding globulin concentrations were not significantly related to serum insulin concentration after adjusting for age and body mass index. Conclusions: We observed increased sex hormone-binding globulin concentration in diabetes man, and was a positively related to age. Further studies are needed to understand the relationships between age, insulin resistance, testosterone, and sex hormone-binding globulin concentrations(J Kor Diabetes Asso 24:699~707,2000).

포도당과 이소프로테레놀에 의한 시상하부의 소마토스타틴 분비가 갑상선자극호르몬방출호르몬에 의한 갑상선자극호르몬에 미치는 효과

박철영,양인명,오승준,김덕윤,우정택,김성운,김진우,김영설,최영길 대한내분비학회 2000 Endocrinology and metabolism Vol.15 No.4·5

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