• Radiation Oncology Journal
• /
• 제21권1호
• /
• pp.54-65
• /
• 2003

목적 : 동시에 대량으로 유전자발현 양상을 검사할 수 있는 cDNA microarray 기법을 이용하여 자궁경부암에서 특징적으로 나타나는 유전자발현 양상을 알아보고, 방사선치료 및 방사선 항암화학요법 병용치료시의 유전자발현 변화양상을 파악하고자 하였다. 대상 및 방법 :자궁경부 편평상피암으로 확진된 후 근치목적 방사선치료를 단독으로 시행한 8명과 항암화학요법을 병행한 8명에서 채취한 종양조직을 대상으로 하고, 정상 자궁경부 3례를 대조군으로 하였다 조직 생검은 치료 전과 외부 방사선치료 16.2$\~$27 Gy에 두 번하였다. 항암화학요법을 병용한 경우, 5-FU 1,000 mg/m$^{2}$을 제 1일부터 5일까지 정주하고, clsplatin 60 mg/m$^{2}$을 제 1일에 정주하였다. cDNA microarray는 종양조직에서 추출한 total RNA를 역전사(reverse transcription)방법을 이용하여 (P-33)을 표지한 cDNAS를 제작, nylon membrane에 hybridization하였다. 이후 membrane을 phosphor-imager screens에 옮겨 1$\~$5일 동안 노출시킨 후 이미지를 스캔하였다. 유전자의 발현정도는 각 스팟(spot)들의 방사능 강도로 나타나는데, 각 스팟의 픽셀(pixel)을 Arrayguage를 사용하여 산출한 후 엑셀파일로 저장하였다. 유전자의 발현정도 비교는 원 자료(original data)를 Z-변환을 통해 보정(normalized)한 후 Z-ratio값을 산출하여 시행하였다. 결과 : 대조군에 비해 자궁경부암에서 Z-ratio 2.0 이상으로 유의한 발현증가를 보인 유전자들은 integrin-linked kinase, CDC28 protein kinase 2, Spry 2, ERK 3 등 15개로 주로 세포성장과 증식, 세포주기, 신호전달 등에 관련된 유전자들이었으며, Z-ratio -2.0 이하의 유의한 발현감소는 G protein-coupled receptor kinase 6외 6개였다. 방사선 단독치료를 시행한 후 Z-ratio 2.0 이상 발현이 증가한 것은 cyclic nucleotlde gated channel외 3개의 Expressed sequence tags (EST)들이었고, Z-ratio -2.0 이하의 발현감소를 보인 것들에는 치료전 종양세포에서 발현이 증가되었던 세포성장과 증식, 세포주기, 신호전달 등에 관련된 유전자들이 포함되었다 방사선치료와 항암화학요법을 병용했을 때는 방사선 단독치료에 비하여 세포성장과 증식 및 신호전달 관련 유전자들이 상대적으로 높게 발현되었으며, 이외에도 혈관형성(angiopoietin-2), 면역반응(formyl peptide receptor-like 1), DNA 손상회복에 관련된 유전자(CAMP phosphodiesterase)의 발현은 증가되고 세포고사(death associated protein kinase)에 관련된 유전자는 발현 감소를 보였다. 결론 : 자궁경부암에서분열과 증식 및 신호전달에 관여하는 여러 종류의 유전자들 발현이 동시다발적으로 증가되어 있다는 것과 방사선치료를 시행하면 이들 유전자의 발현이 감소하여 종양세포의 분열과 증식이 저해된다는 것을 확인하였다. 방사선 단독치료와 항암화학요법 병용치료를 비교하면 그 유전자 발현양상이 다르므로 향후 이번연구에서 나타난 유전자들에 대한 추가 연구가 필요하며, 이는 개별화된 맞춤형 치료법을 개발하는데 기초자료로 사용될 수 있을 것으로 기대된다.

• 한국환경과학회지
• /
• 제16권2호
• /
• pp.219-225
• /
• 2007

Endocrine disrupting compounds (EDC's) are chemicals that either mimic endogenous hormones interfering with pharmacokinetics or act by other mechanisms. Some endocrine disrupters were reported to be chemical substances that cause apoptosis in cells. A number of reports have indicated that 1,3-DCP, one of the EDC's may act as an endocrine disrupter and also has possible carcinogenic effects. 1,3-DCP, present in commercial protein hydrolysates used for human nutrition, are genotoxic and 1,3-dichloro-2-propanol induced tumors in rats. In the present study, it was investigated whether 1,3-DCP induces ROS generation and apotosis in A549 adenocarcinoma cells. Here we show that 1,3-DCP inhibits the growth of lung cancer cell lines and generates reactive oxygen species (ROS), a major cause of DNA damage and genetic instability, It was investigated that 1,3-DCP increases G1 phase cells after 12 hours, thereafter abruptly draws A549 cells to G0 state after 24 hours by flow cytometric analysis. 1,3-DCP induces p53 and $p21^{Cip1/WAF1}$ activation time- and dose-dependently by 24 hours, while the level $p21^{Cip1/WAF1}$ was decreased after 48 hours. These results suggest that 1,3-DCP, an EDC's generates ROS and regulates genes involved with cell cycle and apoptosis.

• 동의생리병리학회지
• /
• 제18권4호
• /
• pp.1120-1128
• /
• 2004

Zinc ion has both essential and toxic effects on mammalian cells. The results demonstrated that the ability of zinc to act as an inducer of apoptosis in C6 glial cells. Incubation with 0.2 mM ZnCl₂ caused cell death that was characterized as apoptosis by internucleosomal DNA fragmentation, formation of apoptotic bodies, nuclear fragmentation and breakdown of the mitochondrial membrane potential. ZnCl₂-induced apoptosis of C6 glial cells was prevented by the addition of Sibjeondaebo-Tang and antioxidants including reduced glutathione, N-acetyl-L-cysteine. We further confirmed that ZnCl₂ decreased the intracellular levels of GSH as well as generation of H₂O₂ in C6 glial cells. In 2D-electrophoresis, computer-assisted comparative analysis of the respective silver stained spot patterns revealed 3 groups with strongly decreased intensity by ZnCl₂. Whereas, 3 groups with increasing intensity were recovered by Sibjeondaebo-Tang. These results suggest that Sibjeondaebo-Tang may function as an antioxidant against free radicals and be applicable to the treatment of brain cells against oxidative stress.

• Parasites, Hosts and Diseases
• /
• 제31권3호
• /
• pp.277-284
• /
• 1993

세계적으로 중요한 열대풍토병 병원체의 하나인 열대리슈마니아의 핵형에 영향을 줄 것으로 기대되는 인자와 그 효과를 관찰하고자 하였다. 토기의 혈액을 포함한 N.N.N. 배지에서 유지하고 있는 열대리슈마니아(Leishmania major)의 promastigote를 열쇽. 약제 첨가 자외선 조사 및 감마선을 이용한 방사선 조사를 여러 가지 방법으로 시행하고 주기변동전기영동(pulsed field gradient gel electrophoresis)을 이용하여 핵형의 변동 여부를 관찰하였다 그 결과 여러 방식에 의한 열쇽과 약제 처리 및 자외선 조사에 따른 핵형의 변화가 없었다 그러나 방사선 조사군에서는 50 Gy 이상 조사한 군에서 1 mesa base pair(Hb) 크기에 있는 염색체부터 소실되기 시작하여 방사선 조사량이 증가함에 따라서 250-500 Kb의 작은 염색체도 파괴되어 500 Gy 이상 군에서는 뚜렷한 염색체 분획이 없이 젤 하단 200 Kb 크기 아래 부분에 몰려 있었다. 이러한 소견은 방사선에 의하여 염색체가 불규칙하게 파손된 것을 의미한다고 하겠다. 방사선을 300 Gy까지 조사한 충체는 계대 배양이 가능하였고. 이들은 원래의 핵형을 유지하였다. 방사선조사 후에 배양된 충체는 염색체가 파괴되지 않았거나 부분적인 손상 후에 DNA 재결합에 의해 원상회복된 것으로 판단된다. 열대리슈마니아의 핵형은 일시적인 자극에 의하여 쉽게 변형되지 않는 안정된 것임을 확인하였다.

• Toxicological Research
• /
• 제20권3호
• /
• pp.213-223
• /
• 2004

This work aimed to study the effectiveness of cellular oxidative parameter (malondial-dehyde, protein carbonyl, and 8-hydroxy-2'deoxyguanosine). The experimental groups were aluminum treated rats and control rats. Aluminum treatd rats were given intraperitoneally aluminum nitrate nonahydrate ($Al^{3+}$, 0.2 mmol/kg) daily for 30 days except Sunday. Control rats were injected 1 ml of saline. After the dose, rats were decapitated and hippocampus and cerebral cortex were removed. The measured parameters were tissue malondialdehyde (MDA, index of lipid peroxidation), protein carbonyl (index of protein oxidation), 8-hydroxy-2'-deoxy-guanosine (8-OHdG, index of DNA oxidation), reduced glutathione (GSH) levels as well as glutathione reductase (GR) and catalase. AI concentrations in the tissues were also measured. All results were corrected by tissue protein levels. The results were as followed; 1. The concentrations of AI in the cortex and hippocampus were significantly higher in the AI-treated rats than in the control rats. 2. Antioxidative enzyme's activity, catalase and GR, were significantly higher in the AI-treated rats than the control rats. GSH levels were also higher in the AI-treated rats. 3. MDA, protein carbonyl, and 8-OHdG concentration of AI-treated rats were significantly higher than those of control rats. 4. The concentrations of antioxidants, and oxidative stress parameter were correlated with the concentrations of AI in hippocampus and cerebral cortex. Catalase and GR activity were also correlated with the concentration of AI. Based on these results, it can be suggested that intraperitoneally injected AI was accumulated in the brain and induced the increase of antioxidant levels and antioxidative enzyme activity. Also, the oxidative products of cellular macromolecules are significantly related to tissue AI concentration. Therefore MDA, protein carbonyl, and 8-OHdG are useful markers for oxidative stress on cellular macromolecules.

• 대한한방내과학회지
• /
• 제19권1호
• /
• pp.22-38
• /
• 1998

This study was to investigate the hepatoprotective and anticirrhotic effects of Ganyeumilhobang(GIE) on the acute and chronic liver injury induced by various agents. Chronic liver injury induced by dimethylnitrosamine(DMN) ; a new experimental model for cirrhosis and the intraperitoneal injection of dimethylnitrosamine in the rat. Acute liver njury induced by carbon tetrachloride$(CCl_4)$ and D-galactosamine ; a experimental model for acute liver injury, the administration of $CCl_4$ and the intraperitoneal injection of D-galactosamine in the rat. The development of fibrosis and acute liver injury by the three prescriptions were examined by the chemical analysis of AST, ALT, prothrombin time and hydroxyproline. The results obtained were as follows. 1. The increasing level of hydroxyproline volume induced by DMN in mice was decreased by the oral administration of GIB. 2. The degree of histological fibrosis and hepatic inflammatory cell infiltration induced by $CCl_4$ decreased by the oral administration of GIB. 3. The increase of senun AST and ALT of mice with acute liver damage induced by $CCl_4$ and D-galactosamine was inhibited by the administration of GIB. 4. The prolongation of prothrombin time(seconds) of mice acute liver damage induced by $CCl_4$ was shortened by the oral administration of GIB. 5. The liver of mice was hepatectomized partial1y after the oral administration of GIB. The mitotic index(% of nuclei), weight of liver, contents of protein, RNA and DNA synthesis of the liver tissue were increased by the oral administration of GIB.

• 대한한방내과학회지
• /
• 제31권1호
• /
• pp.66-78
• /
• 2010

Objective : This study was designed to investigate the possibility of AR for chronic renal injury. Methods : The author first investigated the expression levels of DNA by inducing of chronic renal injury. Then, the author investigated the effects of AR on chronic renal injury induced by combination treatment with Adriamycin and cisplatin in terms of changes in body weights and renal tissues, urine volume, BUN and creatinine levels, creatinine clearance and histopathological changes in renal tissues. Total expression levels of 546 genes were elevated or lowered by induction of chronic renal injury. Genes of which whose expression levels were elevated by induction of chronic renal injury were related to the PPAR signaling pathway and fatty acid mechanism, etc. Genes of which expression levels were lowered by induction of chronic renal injury were related to the neuroactive ligand-receptor signaling pathway. Results : Oral administration of AR restored renal mass which was reduced by induction of chronic renal injury. AR also restored creatinine clearance and lowered serum BUN level. In histopathological observation, the AR group has a tendency to prevent tissue damages as shown in the chronic renal injury group. Conclusions : AR can be used to treat patients with chronic renal injury although further study will be needed to elucidate the exact mechanisms in the efficacy of AR on chronic renal injury.

• 한국수산과학회지
• /
• 제50권6호
• /
• pp.714-720
• /
• 2017

Exposure to ultraviolet (UV) radiation is associated with the development of adverse effects in skin. Among the three types of UV rays, UV-B causes the most damaging effects, inducing sunburn and penetrating the outer skin, resulting in DNA mutations and skin cancer. The objective of this study was to formulate a UV-protective film by incorporating Ecklonia cava extracts. Cells covered with the film were exposed to UV-B (50, 80, and $100mJ/cm^2$). To determine the protective effects of the film, we evaluated cell viability, intracellular ROS generation, and apoptosis. We found that all E. cava extracts absorbed UV light and exhibited protective effects against UV-B-induced photodamage. Among the protective films examined in this study, that incorporating an E. cava 70% ethanol extract (70EX) formed the most effective protection against UV-B in HaCaT cells. These findings suggest that the application of film containing E. cava extract could prevent UV-B-induced photodamage, and offer protection against the detrimental effects of UV radiation, thus maintaining physiological condition.

• 한국수산과학회지
• /
• 제53권1호
• /
• pp.123-131
• /
• 2020

In this study, we investigated the protective effects of a Neutrase enzymatic hydrolysate derived from Korea pen shell Atrina pectinata (APN) against hydrogen peroxide (H₂O₂)-induced oxidative damage in hepatocytes. First, we confirmed that APN has antioxidant activities by scavenging 2,2-azino-bis (3-ethylbenzthiazoline)-6-sulfonic acid radical (ABTS⁺) and H₂O₂ and increasing oxygen radical absorbance capacity (ORAC) value. Also, the treatment of APN increased the cell viability by reducing the intracellular reactive oxygen species (ROS) production in H₂O₂-stimulated hepatocytes. In addition, APN decreased the sub-G₁ DNA contents and the apoptotic body formation increased by H₂O₂ stimulation. Moreover, APN modulated the protein expression of apoptosis related molecules (Bcl-2, Bax and p53) by suppressing the activation of nuclear factor NFkB and ERK/p38 signaling in H₂O₂-stimulated hepatocytes. Furthermore, APN led to the activation of Nrf2/HO-1signaling known as antioxidant systems. These results suggest APN protects hepatocytes against oxidative damages caused by H₂O₂ stimulation.

• 대한한의학회지
• /
• 제23권4호
• /
• pp.140-150
• /
• 2002

Objectives: The water extract of Omija-tang (OMIT) has traditionally been used for treatment of ischemic heart and brain damage in oriental medicine. However, little is known about the mechanism by which the water extract of OMJT rescues cells from these damages. Therefore, this study was designed to investigate the protective mechanisms of OMJT on oxidative stress-induced toxicity in H9c2 cardiomyoblast cells. Methods: Treatments of $H_2O_2$, or $ZnC_{12}$ markedly induced death of H9c2 cardiomyoblast cells in a dose-dependent manner. The characteristics of oxidative stress-induced death of H9c2 showed apparent apoptotic features such as DNA fragmentation. OMJT significantly reduced both ${H_2O_2}-induced$ cell death and chromatin fragmentation. The decrease of B치-XL expression by $H_2O_2$ were inhibited by OMJT. In addition, the increase of Bcl-XS expression was also inhibited by OMJT. In particular, Fas expression, which is generally recognized as cell death-inducing signal by Fas/FasL interaction, was markedly increased by H2O2 in a time-dependent manner. Also, the expression profile of proteins in Chang cells were screened by using two-dimensional (2-D) gel electrophoresis. Among 300 spots resolved in 2-D gels; the comparison of control versus apoptotis cells revealed that signal intensity of 6 spots decreased and 11 spots increased. Results and Conclusions: Taken together, this study suggests that the protective effects of the water extract of OMJT against oxidative damages may be mediated by the modulation of Bcl-XL/S Fas expression.