• Journal of Korean Neurosurgical Society
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• 제38권1호
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• pp.47-53
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• 2005

Objective : Anti-malaria drugs may modulate tumor resistance to chemotherapeutic agents, but it has not been proven effective in the treatment of malignant gliomas. The aim of this study was to determine whether adequate pre-clinical data on co-administration of chemotherapeutic agents with anti-malaria drugs on malignant cell lines could be obtained that would warrant its further potential consideration for use in a clinical trial for malignant gliomas. Methods : Two malignant glioma cell lines [U87MG, T98G] were treated with chemotherapeutic agents alone or with anti-malaria drugs. Cells were incubated with drugs for 4 days. Following the 4-day incubation, drug sensitivity assays were performed using 3-[4,5-dimethyl-2-thiazol-2-yl] 2,5-diphenyltetrazolium bromide [MTT] assay following optimization of experimental conditions for each cell lines and cell viability was calculated. Results : In all of four chemotherapeutic agents[doxorubicin. vincrisitne, nimustine, and cisplatin], the cell viability was found to be markedly decreased when hydroxychloroquine was co-administered on both U87MG and T98G cell lines. The two way analysis of variance[ANOVA] yielded a statistically significant two-sided p-value of 0.0033[doxorubicin], 0.0005[vincrisitne], 0.0007[nimustine], and 0.0003[cisplatin] on U87MG cell lines and 0.0006[doxorubicin], 0.0421[vincrisitne], 0.0317[nimustine], and 0.0001[cisplatin] on T98G cell lines, respectively. However, treatment with chloroquine and primaquine did not induce a decrease in cell viability on both U87MG and T98G cell lines. Conclusion : Our data support further consideration of the use of hydroxychloroquine prior to systemic chemotherapy to maximize its tumoricidal effect for patients with malignant gliomas.

• Journal of Korean Neurosurgical Society
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• 제29권6호
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• pp.731-737
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• 2000

목 적 : 종양세포의 세포면에서는 성장인자 수용체들이 과발현되고 이들이 성장인자들과 결합함으로서 세포의 성장을 촉진하게 된다. 원발성 악성 뇌종양에 대한 새로운 치료법 중 하나가 면역독소를 사용하는 방법이다. 독소는 대상 세포의 세포면에 있는 수용체를 통해 세포질로 이행되며, 면역독소가 선택적인 항종양작용을 나타내기 위해서는 수용체가 과발현되어야 한다. 강력한 세포독성의 효과가 있음에도 불구하고 면역독소를 환자에 투여하였을 때 종양을 완치시키지는 못하였는데, 면역독소의 효과에 영향을 미치는 여러 인자들에 대한 더 많은 연구가 요구된다. 이러한 연구에 사용되어질 뇌종양세포주들에서 성장인자수용체들의 발현 여부를 알아보기 위해 본 연구를 시행하였다. 대상 및 방법 : 한 개의 수모세포종 세포주(Daoy)와 두 개의 교모세포종 세포주(U373 MG, T98 G)에서, transferrin 수용체, insulin-like growth factor-1 수용체, 그리고 interleukin-4 수용체들의 발현을 flow cytometric analysis를 이용하여 조사하였다. 결 과 : Transferrin 수용체와 interleukin-4 수용체는 Daoy, U373 MG, 그리고 T98 G 모두에서 발현되었다. Insulin-like growth factor-1 수용체는 Daoy와 U373 MG에서는 발현되었지만 T98 G에서는 발현되지 않았다. 결 론 : Transferrin 수용체와 interleukin-4 수용체는 면역독소 치료에 적합한 것으로 보인다. 본 실험의 결과는 상기의 세포주를 사용하여 면역독소와 관련된 실험을 하는데 참고가 되어야하며, 면역독소 치료에 있어서 적절한 면역독소를 선택하는 문제 등 치료 모델을 확립하는데 고려되어야 할 것이다.

• 대한한방내과학회지
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• 제28권3호
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• pp.409-420
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• 2007

Objectives : The purpose of this study was to identify the anti-tumor effects of realgar on various cancer cells through molecular biologic and cellular biologic methods. Materials & Methods : We used 5 kinds of cancer cell lines:stomach cancer cell (AGS), glioma cells (T98G, A172, SNU-489) and prostate cancer cells (LNCaP). We injected the boiled extract of realgar. $50{\mu}$g/ml and $100{\mu}$g/ml to culture media (ml) for 24 hours. We examined the morphological changes under an inverted microscope and a fluorescence microscope. We measured the suppressive effect on viability of 5 kinds of cancer cells via XTT assay. We examined the effect on the revelation of PARP cleavage, Bcl-2 protein and Bax protein by western blot analysis. Results : The extract of realgar caused markedly morphological changes on AGS, T98G, SNU-489, and LNCaP. All of them showed withdrawn and floating appearance. The suppressive effect on viability of AGS, T98G, A172, SNU-489, and LNCaP showed that each test group had more suppressive effect on viability of AGS, T98G, A172, SNU-489, and LNCaP than the control group, which was statistically significantly (p<0.01). The extract of realgar did not induce PARP cleavage in AGS, T98G, A 172, SNU-489, or LNCaP. In the revelation of protein related to apoptosis, the protein levels of Bcl-2 decreased and the protein levels of Bax increased in AGS, T98G, SNU-489, and LNCaP treated with realgar. The protein levels of Bcl-2 decreased and the protein levels of Bax did not change in A172 treated with realgar. Conclusions : This experiment showed that realgar has anti-tumor effect on stomach cancer cells (AGS), glioma cells (T98G, SNU-489L and prostate cancer cells (LNCaP)

• 생명과학회지
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• 제17권3호통권83호
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• pp.381-385
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• 2007

FACT(facilitates chromatin transcription)은 크로마틴을 주형으로 하는 전사에 필요한 크로마틴 특이적 전사 진행 인자이다. FACT는 Saccharomyces Cerevisiae Spt16/Cdc68의인간 유사체와 high mobility froup-1-like protein structure-specific recognition protein-1(SSRP-1)의 이종단백질 복합체이다 본 논문에서는 FACT의 단위체인 hSpt16의 발현이 휴지기의 T98G 세포에서 급격히 감소됨을 면역형광 분석법과 Western blot 분석법을 이용하여 관찰하였다. 이와 반대로 증식기의 T98C 세포에서는 hSpt16이 높은 수준으로 발현되고 있는 것이 관찰되었다. FACT의 또 다른 단위체 SSRP-1의 발현은 휴지기나 증식기의 세포에서 변화가 없음이 관찰되었다. 이상의 결과로부터 hSpt16이 발현이 세포의증식과 연관이 되어 있음을 알 수 있었고 세포 증식 표지 인자로 사용될 수 있을 것으로 사료된다.

• Journal of Korean Neurosurgical Society
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• 제60권2호
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• pp.130-137
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• 2017

Objective : Autophagy is one of the key responses of cells to programmed cell death. Memantine, an approved anti-dementia drug, has an antiproliferative effect on cancer cells but the mechanism is poorly understood. The aim of the present study was to test the possibility of induction of autophagic cell death by memantine in glioma cell lines. Methods : Glioma cell lines (T-98 G and U-251 MG) were used for this study. Results : The antiproliferative effect of memantine was shown on T-98 G cells, which expressed N-methyl-D-aspartate 1 receptor (NMDAR1). Memantine increased the autophagic-related proteins as the conversion ratio of light chain protein 3-II (LC3-II)-/LC3-I and the expression of beclin-1. Memantine also increased formation of autophagic vacuoles observed under a transmission electron microscope. Transfection of small interfering RNA (siRNA) to knock down NMDAR1 in the glioma cells induced resistance to memantine and decreased the LC3-II/LC3-I ratio in T-98 G cells. Conclusion : Our study demonstrates that in glioma cells, memantine inhibits proliferation and induces autophagy mediated by NMDAR1.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2001년도 춘계학술발표대회
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• pp.69-69
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• 2001

The cell cycle phase in which donor nuclei exist prior to nuclear transfer is an important factor governing developmental rates of reconstituted embryos. It was suggested that quiescent G0 and cycling G1 cells could support normal development of reconstituted embryos. In a quest of optimized donor nuclei treatment prior to nuclear transfer, this study was undertaken to examine the cell cycle characteristics of bovine fetal and adult somatic cells when cultured under a variety of culture treatments and the cell cycle change with the lapse of time after trypsinization. This was archived by measuring the DNA content of cells using flow cytometry, Cultured fetal fibroblast cells, adult skin and muscle cells, and cumulus cells were divided by 3 culture treatments; 1) grown to 60-70% confluency (cycling), 2) serum starved culture, 3) culture to confluency. Trypsinized cells were fixed by 70% ethanol and stained with propidium iodide. For one experiment, trypsinized cells were resuspended in DMEM＋10% FBS and incubated for 1.5, 3 and 6 h with occasional shaking before ethanol fixation. Cell cycle phases were determined by flow cytometry enabling calculation of percentages of G0＋G1, S and G2＋M. The majority of cells were in G0＋Gl stage regardless of origin of cells. Cultures that were serum starved or cultured to confluency contained significantly (P<0.05) higher percentages of cells in G0＋G1 (89.5-95.4%). For every cell lines and culture treatments, percentages of cells in existing in G0＋G1 increased with decreasing of the cell size from large to small. In the serum starved and confluency groups, about 98% of small cells were in G0＋G1 Serum starved culture contained higher percentages of small-sized cells (38.5-66.9%) than cycling and confluent cultures regardless of cell lines (P<0.05). After trypsinization of fetal fibroblast and adult skin cells that were serum starved and cultured to confluency, the percentages of cells in G0＋G1 significantly increased by incubation for 1.5(95.7-99.5%) and 3.0 h (95.9-98.6%). The results suggest that the efficient synchronization of bovine somatic cells in G0＋G1 for nuclear transfer can be established by incubation for a limited time period after trypsinization of serum starved or confluent cells.

• IMMUNE NETWORK
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• 제5권2호
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• pp.89-98
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• 2005

Background: DNA vaccination represents an anticipated approach for the control of numerous infectious diseases. Used alone, however, DNA vaccine is weak immunogen inferior to viral vectors. In recent, heterologous prime-boost vaccination leads DNA vaccines to practical reality. Methods: We assessed prime-boost immunization strategies with a DNA vaccine (minigene, $gB_{498-505}$ DNA) and recombinant vaccinia virus $(vvgB_{498-505})$ expressing epitope $gB_{498-505}$ (SSIEF ARL) of CD8+ T cells specific for glycoprotein B (gB) of herpes simplex virus (HSV). Animals were immunized primarily with $gB_{498-505}$ epitope-expressing DNA vaccine/recombinant vaccinia virus and boosted with alternative vaccine type expressing entire Ag. Results: In prime-boost protocols using vvgBw (recombinant vaccinia virus expressing entire Ag) and $vvgB_{498-505}$, CD8+ T cell-mediated immunity was induced maximally at both acute and memory stages if primed with vvgBw and boosted with $vvgB_{498-505}$ as evaluated by CTL activity, intracellular IFN-staining, and MHC class I tetramer staining. Similarly $gB_{498-505}$ DNA prime-gBw DNA (DNA vaccine expressing entire Ag) boost immunization elicited the strongest CD8+ T cell responses in protocols based on DNA vaccine. However, the level of CD8+ T cell-mediated immunity induced with prime-boost vaccination using DNA vaccine expressing epitope or entire Ag was inferior to those based on vvgBw and $vvgB_{498-505}$. Of particular interest CD8+ T cell-mediated immunity was optimally induced when $vvgB_{498-505}$ was used to prime and gB DNA was used as alternative boost. Especially CD7+ T cell responses induced by such protocol was longer lasted than other protocols. Conclusion: These facts direct to search for the effective strategy to induce optimal CD8+ T cell-mediated immunity against cancer and viral infection.

• 한국식품영양학회지
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• 제28권6호
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• pp.965-972
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• 2015

This study was conducted to evaluate the neuroprotective effects of Cheonggukjang extract in in-vitro and in-vivo models. T98G-human glioblastoma cells were pretreated with various concentrations (1~10 mg/mL) of Cheonggukjang extract for 24 h and then exposed to $H_2O_2$ (1 mM) for 3 h. The neuroprotective effects of Cheonggukjang extract were measured using a CCK-8 kit assay, total antioxidant capacity (TAC) assay, reactive oxygen species (ROS) assay, and lactate dehydrogenase (LDH) release assay. The early stage focal ischemia rodent model was used as the in-vivo neurotoxicity model. Various concentrations (10~200 mg) of Cheonggukjang extract were administered to the animal models for 1 week. Peripheral blood was analyzed for glutathione peroxidase (GPx) expression by ELISA, and infarct volume reduction was analyzed by TTC staining. Cheonggukjang extract significantly (p<0.05) increased cell viability in T98G cells against $H_2O_2$ as well as against the induced neurotoxicity. Indeed, treatment with the Cheonggukjang extract induced a decrease in ROS and LDH expression and increased TAC significantly (p<0.05). However, Cheonggukjang extract did not induce a decrease in infarct volume or an increase in GPx expression in the in-vivo model. Despite the limitation in neuroprotection, Cheonggukjang extract may be useful for treating ROS injury.

• The Plant Pathology Journal
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• 제21권4호
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• pp.301-309
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• 2005

Trichoderma species/isolates exhibited varied degree of agglutination on sclerotial (Sc) and hyphal (Hy) surface of Macrophomina phaseolina. The agglutination efficiencies on Sc and Hy ranged from $11\;to\;57\%$. Isolates of T. harzianum (Th) and T. viride (Tv) showed greater agglutination on Sc ($23-57\%$) and Hy ($16-47\%$). Different enzymes (trypsin, pepsin, proteinase k, a-chymotrypsin, lyticase and glucosidase) and inhibitors (tunicamycin, cycloheximide, brefeldin A, sodium azide, dithiothreitol and SDS) reduced the agglutination potential of conidia of Th-23/98 and Tv-25/98; however, the extent of response varied greatly in different treatments. Different fractions of Th-23/98 and Tv-25/98 exhibited haemagglutinating reaction with human blood group A, B, AB and O. Haemagglutinating activity was inhibited by different sugars and glycoproteins tested. Crude haemagglutinating protein from outer cell wall protein fraction of Th-23/98 and Tv-25/98 were eluted on Sephadex G-100 column. Initially Th-23/98 and Tv-25/98 exhibited two peaks showing no agglutination activity; however, lectin activity was detected in the third peak. Similar to crude lectin, the purified lectin also exhibited haemagglutinating activity with different erythrocyte source. SDS-PAGE analysis of partially purified lectin revealed single band with an estimated molecular mass of 55 and 52 kDa in Th-23/98 and Tv-25/98, respectively. Trypsin, chymotrypsin and b-1,3-glucanase totally inhibited lectin activity. Similarly, various pH also affected the haemagglutinating activity of Th-23/98 and Tv-25/98. From the present observations, it can be concluded that the recognition/attachment of mycoparasite (T. harzianum and T. viride) to the host surface (M. phaseolina) may be most likely due to lectin-carbohydrate interaction.

• 대한화장품학회지
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• 제41권2호
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• pp.173-180
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• 2015

본 연구에서는 항산화 기능을 가진 자소엽, 어성초 그리고 녹차 추출물들과 이들 추출물의 혼합물을 사용하여 항산화, 미백, 세포독성실험을 진행하였다. 자소엽, 어성초와 녹차의 추출물을 동일 양으로 혼합하였을 때 처리농도가 증가함에 따라 농도 의존적으로 DPPH와 ABTS 라디칼 소거활성이 높아졌으며, 추출물을 혼합한 경우 낮은 농도인 $10{\mu}g/mL$에서도 80.2%, 98.0%의 DPPH와 ABTS 라디칼을 소거시키는 우수한 항산화 효과를 나타내었다. 미백효과의 정도를 조사하기 위하여 tyrosinase 저해효과를 조사한 결과 자소엽과 녹차추출물 그리고 각 추출물의 혼합물이 농도 의존적으로 tyrosinase 저해효과가 증가되었다. HaCaT 세포를 사용한 세포독성실험의 경우 자소엽, 어성초 추출물의 경우 세포생존율이 무처리 대조군과 같거나 높게 나타났으며 특히 어성초추출물의 경우 $100{\mu}g/mL$에서 무처리 대조군 보다 10% 이상의 높은 생존율을 보였다. 녹차와 각 식물 추출물 혼합물의 경우 $500{\mu}g/mL$ 이상에서는 무처리 대조군에 비하여 세포생존율이 감소하였다. 그러므로 세 가지 식물의 추출물 혼합물이 $100{\mu}g/mL$ 농도일 때 세포에 안전하며 높은 항산화 활성과 함께 tyrosinase 저해 효과가 있었다.