Desferrioxamine이 사람 간암 세포주의 DNA 합성에 미치는 영향

강진경(Jin Kyung Kang),김원호(Won Ho Kim),송시영(Si Young Song),김도영(Doe Young Kim),문일환(Il Hwan Moon),윤견일(Kyun Il Yoon) 대한소화기학회 1993 대한소화기학회지 Vol.25 No.6

N/A Desferrioxamine (DFO), an iron chelator, has been sbown to have antiproliferative activity in a variety of malignant cells including hepatocellular carcinoma. The antiproliferative effect of DFO's known to be caused by decreased activity of ribonucleotide reductase, a key enzyme in DNA synthesis. This study was conducted to investigate the effect Of DFO on the DNA synthesis of cultured hepatoma cells. The proliferation of hepatocellular carcinoma (Hep 3B) as well as hepatoblastoma (Hep G2) cells was measured by trypan blue dye exclusion method and the DNA synthesis was measured by [3H] thymidine incorporation. The results obtained were as follows: The proliferation of hepatoma cells was slightly inhibited by 2 ug/ml and markedly inhibited by 6 ug/ml of DFO. This antiproliferative effect was not enhanced any more by higher dose of DFO. The percent viability of Hep 3B and Hep G2 cells was above 90%. after 96 hours of incubation with 60 ug/ml of DFO and that of Hep 3B and Hep G2 cells was 88.0% and 89.6% respectively after l6 hours of culture with 120 ug/ml of DFO. DNA synthesis of hepatoma ceils was decreased by DFO in a dose dependent manner up to 20 ug/ ml. The decrease of DNA synthesis was not enhanced any more by higher dose of DFO. In conclusion, the antiproliferative effect of DFO on cultured human hepatoma cell lines was caused by the inhibition of DNA synthesis rather than by direct cytocidal effect.

Desferrioxamine이 사람 간암 세포주의 세포주기에 미치는 영향

강진경(Jin Kyung Kang),박인서(In Suh Park),김원호(Won Ho Kim),김도영(Doe Young Kim),권오헌(Oh Hun Kwon),홍성근(Seong Keun Hong),이정운(Jung Woon Lee) 대한소화기학회 1993 대한소화기학회지 Vol.25 No.6

N/A Desferrioxamine (DFO), an iron chelator, has been known to have an antiproliferative activity in a variety of malignant cell lines including human hepatoma cell lines. This study was conducted to investigate the effect of DFO on the cell cycle of hepatoma cells. Using Hep 3B cells as the hepatoma cell lines, bivariate DNA flow cytometry, after staining with propidium iodide and antibromodeoxyuridine following bromodeoxyuridine labelling, was performed to analyze the cell cycle. The following results were obtained. The proportion of S phase cells increased and that of GO/Gl phase cells decreased after the addition of DFO in the culture media in a dose dependent manner up to 20 ug/ml. The S phase duration (Ts) was 9.9 hours when cultured without DFO, but after the addition of 20 pg/ml of DFO, the increment in the proportion of early-S phase continued without shift to the mid-S and late-S phase, and Ts was markedly prolonged (54.1 hours). After removal of DFO from the culture media, a sequential increase from early-S through mid-S and late-S to G2/M phase was observed. In conclusion, the antiproliferative effect of DFO on cultured human hepatoma cell lines was caused by a block in the early-mid S interface or mid S phase of the cell cycle.

Inhibitory Effect of Quercetin and Desferrioxamine in Rat Reflux Esophagitis

Hyun Ju Song,Bong Jin Kil,Ill Woong Kim,Young Sil Min,Dong-Seok Kim,Uy Dong Sohn 대한생리학회-대한약리학회 2001 The Korean Journal of Physiology & Pharmacology Vol.5 No.4

<P> This study was aimed to evaluate the effects of quercetin and desferrioxamine on the development of the reflux esophagitis induced surgically, on gastric secretion and on lipid peroxidation which is a marker of oxidative stress. Omeprazole was used as a positive control drug. Omeprazole significantly and dose-dependently prevented the development of reflux esophagitis, but quercetin or desferrioxamine prevented only at high dose. Omeprazole significantly and dose-dependently inhibited the gastric acid secretion (gastric volume, pH and acid output), but quercetin or desferrioxamine did not inhibit. Malonyldialdehyde content, the end product of lipid peroxidation, increased significantly after the induction of reflux esophagitis. Omeprazole prevented lipid peroxidation. Quercetin and desferrioxamine inhibited the lipid peroxidation independent of their actions on gastric secretion. This result indicates that omeprazole confirmed preventing effect of rat reflux esophagitis, but quercetin and desferrioxamine inhibited esophagitis by reduction of lipid peroxidation irrespective of gastric acid secretion.

지속적 혈액투석환자에서 Desferrioxamine 투여 후의 혈장 Endothelin 농도 변화

최규복(Gyu Bog Choi),윤견일(Kyun Ill Yoon) 대한내과학회 1996 대한내과학회지 Vol.51 No.3

N/A Objectives: It has been reported that the risk of oxygen radical injury is increased in chronic renal failure due to the decreased endogenous serum antioxidants. Especially, the C5a induced by membrane bioincompatibility can stimulates neutrophils to release of oxygen free radicals, resulting in endothelial cell injury. However, Desferrioxamine(DFO) act as an iron chelator, which blocks iron-catalyzed Haber-Weiss reaction and inhibits release of oxygen free radicals from activated neutrophils. It is also reported that endothelin(ET) can be released from endothelial cells in response to vascular damage such as atherosclerosis. Therefore, we administered DFO, into maintenance hemodialyein patients. Then we examined the possibility of oxygen radical injury during interdialytic period and its relation with the plasma ET concentration. Methods: During the last 1-2 hours of hemodialysis, DFO(40mg/kg in 5% D/W 200cc) was infused intravenously into 13 patients(DFO group), and placebo(5% D/W 200cc only) was infused with same manner into 9 patients(Placebo group). We sampled blood for measurement of plasma ET concentration just before the initiation of hemodialysis on the day of infusion, on the 2nd-3rd day and on the 7th day after infusion. Also, we examined 26 non-diabetic patients with normal renal function as a norma1 control. Results: The mean plasma ET concentration in total hemodialysis patients is higher (5.08±3.09pg/ ml) than in normal control (2.58±1.08pg/ml, p<0.01). There was no statistical difference between two hemodialysis groups in plasma ET concentration measured before infusion (5.56±3.50pg/ml in DFO group, 4.38±2.40 pg/ml in placeb group). In DFO group, plasma ET concentration decreased significantly on the 2nd-3rd day (3,49±2.08pg/ml, p<0.01), but increased significantly on the 7th day (5.62± 2.95pg/ml, p<0.05), In contrast, there were no significant changes in plasma ET concentration in placebo group. There was no significant difference in the decrement of plasma ET between the cases of transferrin saturation below and above 60% and there was no relation between the plasma ET decrement and transferrin saturation or serum ferritin in DFO group. Conclusion: The decrease of plasma ET concentration after DFO infusion might be the result of diminished endothelial cell injury from oxygen free radicals. Therefore, we believe that the oxygen radical injury can occur during not only the hemodialysis but also the interdialytic period. Also these results suggest that the oxidant damage of endothelial cell may be one of the causes of elevated ET concentration in chronic renal failure, However, we could not confirm in this study whether the obtained results were caused by the chronic effects of membrane bioincompatibility or by the decreased endogenous serum antioxidants.

배양된 위암세포에 대한 Desferrioxamine 및 항암제의 병합 효과

강진경(Jin Kyung Kang),최흥재(Heung Jai Choi),박인서(In Suh Park),문영명(Young Myoung Moon),김원호(Won Ho Kim),이관식(Kwan Sik Lee),김현숙(Hyon Suk Kim),권오헌(Oh Hun Kwon) 대한소화기학회 1993 대한소화기학회지 Vol.25 No.1

N/A Iron is essential for the growth of all cells including tumor cells because ribonucelotide reductase, a rate-limiting enzyme for DNA synthesis, requires the continual presence of oxygen and iron. Furthermore, it was reported that iron depletion causes tumor cell death but shows little cytotoxic effect on normal human diploid cells in vitro. Desferrioxamine (DFO), an iron-chelating agent, has also been reported to have antitumor activity on several cultured human cancer cell lines but has minimal cytotoxicity on normal human cells. Therefore, it was hypothesized that I)FO could be used as a potential anticancer regimen. This study was designed to investigate the combined effect of DFO and several an.ticancer chemotherapeutic agents, such as 5-fluorouracil (5-FU), adriamycin (ADR), and cisplatinum (DDP), against gastric cancer cell line (PHB). We measured cytotoxicity by MTT assay and evaluated the effects of the combinations by isobologram analysis. The results obtained were as foliows: IC50 of DFO, 5-FU, ADR, and DDP were 14.3 ug/mL, 1.2 ug/mL, 0.5 ug/mL, and 2.9 ug/mL, respectively. The combination of DFO with 5-FU showed a synergistic effect, but combination of DFO with ADR and DFO with DDP did not. We concluded that DFO, which showed an in vitro anticancer effect against cultured gastric cancer cells and showed a synergistic effect if combined with 5-FU, could be used as a potentiaI anticancer drug.

혈액 투석 환자에서 나타나는 rHuEPO 저항성 빈혈에 대한 Desferrioxamine 의 장기 효과

윤경우,배성화,정항재,도준영,임상우 영남대학교 의과대학 1997 Yeungnam University Journal of Medicine Vol.14 No.2

신 부전 환자에게 발생하는 빈혈의 원인은 여러 가지가 있다. 그러나 rHuEPO를 투여할 시 이러한 요인들을 극복하고 빈혈을 교정할 수가 있다. 하지만 약 10%의 환자들에서는 고용량의 rHuEPO에도 불구하고 rHuEPO 저항성 빈혈을 나타낸다. 이러한 경우 DFO는 이러한 저항성을 극복할 수 있다고 하며, 이에 대한 기전은 아직 논란이 많다. 따라서 저자들은 DFO를 투여시 나타나는 여러 인자들을 고려해보고 기존의 보고보다 장기적인 관찰을 통하여 이러한 환자에게서 DFO가 빈혈을 교정할 수 있을지 여부를 알아보고 DFO가 rHuEPO 저항성 빈혈을 극복하는 기전을 추측해보고자 하였다. 영남대학교 의과대학 부속병원 투석실에서 투석을 시행하는 만성 신부전 환자 중 rHuEPO의 효과에 저항성을 나타내는 7명의 환자가 실험군(DFO+EPO)으로 정해지고, 다른 7명이 대조군으로 선정되었다. 실험군은 주당 4000 U의 고용량이 rHuEPO에도 불구하고 혈색소치가 9 g/dL이하 이며 출혈이나 철결핍성 빈혈같은 뚜렷한 원인 없이 정구성 정색소성 빈혈을 나타낸 환자들이었으며 대조군은 나이, 평균 투석 기간에 있어서 대조군과 차이를 보이지 않으며 rHuEPO에 비교적 적절한 반응을 보이는 환자들이었다. DFO는 8주간 rHuEPO와 함께 투여 되어 졌고 rHuEPO의 사용 용량은 지난 6개월의 평균 용량과 같게 투여 되었다(평균 사용 용량은 123.5 U/Kg/Wk). 8주간 DFO가 투여된 후 15개월간 혈색소치와 rHuEPO 사용 용량을 추적하였다. 이 15개월간은 각 개인의 혈색소치와 경제 사정에 의해 rHuEPO의 용량이 결정지어 졌다. 대조군에서는 같은 기간 실험군에서와 같은 방법으로 rHuEPO가 투여되어 졌으나 DFO는 사용되지 않았다. DFO가 투여된 15 개월의 추적기간은 Time I (DFO투여후 7개월간)과 Time II(Time I 이후 8개월간)으로 나누었다. 성적은 다음과 같다 : DFO 투여 전, 실험군의 평균 혈색소치는 대조군과(평균 8.2 g/dL) 같은 정도의(p>0.05) 수준인 7.8 g/dL 이었다. 하지만 rHuEPO의 사용 용량은 평균 123.5 U/Kg/Wk으로서 대조군의 그것(평균 41.6 U/Kg/Wk)보다 의미있게 많은(p<0.05) 용량을 사용하여 rHuEPO에 대한 저항성을 나타내었다. 하지만 DFO사용후 실험군에서 혈색소치는 의미있게 상승되었고(p<0.05) 이것은 Time II에 이르기 까지 지속되었다(Time I : 평균 8.6 g/dL, Time II : 평균 8.6 g/dL). 즉, 혈색소치에 대한 DFO의 효과는 투여 후 15 개월간 Time I, Time II의 시기에 걸쳐 균등한 정도로 지속되었다. 또한 실험군에서 rHuEPO의 사용 용량은 DFO투여후 대조군과 유사한 수준으로 감소하였고 이것도 마찬가지로 15개월간 지속되었다(Time I ;: 평균 48.1 U/Kg/Wk, Time II : 평균 51.8 U/Kg/Wk). 한편, 같은 기간 대조군에서의 혈색소치와 rHuEPO사용 용량의 변화는 없었다. 특히, 실험군에서 혈색소치의 증가와 rHuEPO의 사용 용량의 감소는 DFO투여 후 1개월동안에 극대점을 이루었다. 즉, DFO 사용 후 감소된 rHuEPO사용 용량으로도 조혈 작용을 증강시킬 수 있었고 따라서 DFO를 사용함으로써 rHuEPO의 저항성을 극복할 수 있으리라고 생각된다. 보다 대규모의 실험과 잠재적으로 위험한 DFO의 부작용을 최소화 할 수 있는 적절한 용량과 투여 방법에 대한 연구가 더 필요하다고 사료된다. There are several factors concerning to anemia in chronic renal failure patients. But when rHuEPO is used, most of these factors can be overcome. and the levels of hemoglobin are increased. However, about 10% of the renal failure patients represent rHuEPO-resistant anemia eventhough high dosage of rHuEPO. For these cases, desferrioxamine can be applied to correct rHuEPO resistnacy, and many mechanism of DFO are arguing. So we are going to know whether DFO can be applied to correct anemia of the such patients, how long its effect can be continued. The seven patients as experimental group(DFO+EPO) who represent refractoriness to rHuEPO and the other seven patients as control group(EPO) were included. Experimental group had lower than 9 g/dL of hemoglobin levels despite high rHuEPO dosage (more than 4000U/Wk) and showed normocytic normochromic anemia. There were no definitve causes of anemia such as hemorrhage or iron deficiency. Control group patients had similar characteristics in age, mean dialysis duration but showed adequate response to rHuEPO. DFO was adminsitered to experimental group for 8 weeks along with rHuEPO(the rHuEPO individual mean dosage had been determined by mean dosage of the previous 6 months. Total mean dosage; 123.5 U/Kg/Wk). After 8 weeks of DFO administration, the hemoglobin and rHuEPO dosage levels were checked for 15 consecutive months. It should be noted that the patients determined their own rHuEPO dosage levels according to hemoglobin levels and economic status. In conrol group, rHuEPO was administered by the same method used in experimental group without DFO through the same period. Fifteen months of observation period after DFO trial were divided as Time Ⅰ(7 months after DFO trial) and Time Ⅱ(8months after Time Ⅰ). The results are as follows: Before DFO trial, mean hemoglobin level of experimental group was 7.8g/dL, which is similar level(p>0.05) to control group(mean Hb; 8.2 g/dL). But in experimental group, significantly(p<0.05) higher dosages of rHuEPO(mean; 123.5 U/Kg/Wk) than control group (mean; 41.6 U/Kg/Wk) had been used. It means resistancy to rHuEPO of experimental group. But after DFO trial, the hemoglobin levels of the experimental group were increased significantly(p<0.05), and these effect were continued to Time Ⅱ.(Time Ⅰ; mean 8.6g/dL, Time Ⅱ; mean 8.6g/dL) The effects of DFO to hemoglobin were continued for 15 months after DFO trial with similar degree through Time Ⅰ, Time Ⅱ. Also rHuEPO dosages used in the experimental group were decreased to similar levels of the control group after DFO trial and these effect were also continued for 15 months(Time Ⅰ; mean 48.1 U/Kg/Wk. Time Ⅱ; mean 51.8 U./Kg/Wk). In the same period, hemoglobin levels and rHuEPO dosages used in the control group were not changed significantly. Notibly, hemoglobin increment and rHuEPO usage decrement in experimental group were showed maxilly in the 1st month after DFO trial. That is,after the use of DFO, erythopoiesis was enhanced with a reduced rHuEPO dosage. So we think rHuEPO reisistancy can be overcome by DFO therapy. In conclusion, the DFO can improve the anemia caused by chronic renal failure at least over 1 year, and hence, can reduce the dosage of rHuEPO for anemia correction. Additional studies in order to determine the mechanism of DFO on erythropoiesis and careful attention to potential side effects of DFO will be needed.

Dissolution of Phosphate-adsorbed Goethite by Desferrioxamine B

Priscila Ung,Jang-Hung Huang 한국토양비료학회 2014 한국토양비료학회 학술발표회 초록집 Vol.2014 No.6

Screening of Differentially Expressed Genes by Desferrioxamine or Ferric Ammonium Citrate Treatment in HepG2 Cells

Park, Jong-Hwan,Lee, Hyun-Young,Roh, Soon-Chang,Kim, Hae-Yeong,Yang, Young-Mok Korean Society for Biochemistry and Molecular Biol 2000 Journal of biochemistry and molecular biology Vol.33 No.5

A differential display method is used to identify novel genes whose expression is affected by treatment with ferric ammonium citrate (FAC) or desferrioxamine (DFO), an iron chelating agent in the human hepatoblastoma cell line (HepG2). These chemicals are known to deplete or increase the intracellular concentration of iron, respectively. Initially, we isolated seventeen genes whose expressions are down- or up regulated by the treatment of the chemicals, as well as their four differentially expressed genes that are designated as clone-1, -2, -3, and -4. These are further characterized by cDNA sequencing and Northern blot analysis. Through the cDNA sequencing, as well as comparing them to genes published using the NCBI BLAST program, we identified the sequence of the clone-1 that is up-regulated by the treatment of DFO. It is identical to the human insulin-like growth factor binding protein-1 (IGFBP-1). This suggests that the IGFBP-1 gene in the HepG2 cell is up-regulated by an iron depletion condition. Also, the expression of the clone-3 and -4 is up-regulated by FAC treatment and their eDNA sequences are identical to the human ferritin-fight chain and human NADH-dehydrogenase, respectively. However, the sequence of the clone-2 has no significant homology to any other known gene. Therefore, we suggest that changes of the cellular iron level in the HepG2 cell affects the transcription of cellular genes. This includes human IGFBP-1, ferritin-fight chain, and NADH-dehydrogenase. Regulation of these gene expressions may have an important role in cellular functions that are related to cellular iron metabolism.

Hypoxia Enhances Osteogenic Differentiation in Retinoic Acid-Treated Murine-Induced Pluripotent Stem Cells

Jeeranan Manokawinchoke,Thanaphum Osathanon,Hiroshi Egusa,Prasit Pavasant 한국조직공학과 재생의학회 2016 조직공학과 재생의학 Vol.13 No.5

Hypoxic condition influences biological responses in various cell types. However, a hypoxic regulating osteogenic differentiation remains controversy. Here, an influence of short-term culture in hypoxic condition on osteogenic marker gene expression by retinoic acid-treated murine gingival fibroblast-derived induced pluripotent stem cells (RA-miPS) was investigated. Results demonstrated that hypoxic condition significantly upregulated Vegf, Runx2, Osx, and Ocn mRNA expression by RA-miPS in normal culture medium at day 3. Further, desferrioxamine significantly downregulated pluripotent marker (Nanog and Oct4) and enhanced osteogenic marker (Runx2, Osx, Dlx5, and Ocn) gene expression as well as promoted in vitro mineral deposition. However, the effect of cobalt chloride on osteogenic differentiation of RA-miPS was not robust. In summary, the results imply that hypoxic condition may be useful in the enhancement of osteogenic differentiation in RA-miPS.

단핵구의 부착, 분화 및 SR-A 발현에 대한 철 결핍의 영향

서검석(Geom Seog Seo),최석채(Suck Chei Choi),최은영(Eun Young Choi),김태현(Tae Hyeon Kim),나용호(Yong Ho Nah),오재민(Jae Min Oh),김상욱(Sang Wook Kim),전창덕(Chang-Duk Jun) 대한해부학회 2004 Anatomy & Cell Biology Vol.37 No.6

철 항산성의 유지는 세포의 증식 및 분화에 필수적이며, 면역 기능 조절에도 중요한 역할을 한다. 단핵구와 큰포식세포는 급만성 염증에 있어 숙주 방어의 중요 역할을 한다. 또한 단핵구 및 큰포식세포는 세망내피계통의 다른 세포들과 더불어 철의 저장에 있어 중요한 역할을 한다. 이러한 단핵구인 THP-1 세포에서 철 결핍이 발생될 때 형태학적 및 기능적 변화에 대한 연구가 없어 알아보았다. Desferrioxamine (DFO)을 처리한 단핵구에서는 세포 부착능이 증가하였고 미분간섭 현미경(Differential interference contrast, DIC)에서 DFO를 처리한 세포의 거짓발 연장(pseudopodial extension)이 관찰되었다. Giemsa 염색에서 핵세포질 비율(N/C ratio)의 변화를 보였다. 또한 fluorescence-activated cell sorter (FACS)와 rhodamine-phalloidin 염색에서 DFO에 의한 단핵구의 탐식능 증가가 관찰되였다. 세포 분화 인자인 scavenger receptor-A (SR-A)의 발현이 DFO 처리한 단핵구에서 증가되었다. 이러한 결과는 DFO에 의한 철 결핍 상태가 단핵구의 큰포식세포양 세포로 분화를 촉진시키며, 이는 염증에서 숙주 면역 반응을 약화시킬 것으로 추측된다. Maintenance of cellular iron homeostasis is a prerequisite for proliferation and differentiation of cells, and is also a central role in the regulation of immune function. Monocyte-macrophages play an important roles in host defense, particularly in the inflammatory process of acute and chronic disease. The reason that an iron is important in these cell is because an iron is indispensable in a generation of hydroxyl radical for bacterium killing. Because of the role of iron in the monocytic THP-1 cell differentiation is not become clear, we investigated whether THP-1 cell can differentiate to macrophage-like cell using of iron and iron chelator which cause iron depletion. The cell differentiation was not able to observe by iron treatment, by the way, the cell adhesion was increased in DFO treated monocyte and cellular pseodopodial extension, change of a nucleus-cytoplasmic ratio were showed in Differential interference contrast (DIC) and Giemsa staining, and it was inhibited by ferric citrate (FC). Increased polystyrene bead phagocytosis by DFO treatment of THP-1 cell were detected through FACS and rhodamine-phallodin staining. The SR-A expression, which was a cell differentiation marker, was increased by DFO treatment of THP-1 cell. These results suggest that iron depletion by DFO can promote THP-1 cell diffentiation into macrophage-like cell, and this may carrying out important role in the immune response.