• 한국식품영양과학회지
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• 제42권7호
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• pp.1022-1028
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• 2013

러시아에서 여러 민간요법으로 사용되어지고 있는 차가버섯은 대장암, 위암 등에서 항암효능이 있다고 알려져 있다. 그러나 뇌종양에 대한 연구는 현재 이루어지지 않고 있으며 차가버섯의 항암 효능에 대한 기전 연구도 미미한 실정이다. 따라서 본 연구에서는 신경교모세포종 U-87 MG 세포주에 대한 성장억제 효과를 확인하고자 세포생존율 변화와 형태학적 변화, 세포주기에 미치는 영향과 세포사멸 관련 단백질 및 세포주기 조절 단백질의 발현 변화를 조사하였다. 그 결과 차가버섯 추출물을 처리한 U-87 MG 세포 생존율이 유의하게 감소하였으며, Bcl-2, Bax와 p-p53 단백질 수준차이는 없었고 caspase-3 활성에 영향을 미쳐 죽음 수용체 경로에 의한 세포사멸이 이루어졌다. 또한 세포주기를 관찰한 결과 $G_0/G_1$ 세포주기에서 성장 억제를 확인하였으며, 이는 cyclin D1, CDK2 단백질 발현량의 감소와 RB 단백질의 인산화 억제 때문이라는 결론에 도달하였다. 본 연구결과는 차가버섯을 뇌종양의 치료 및 보조제로서 역할을 수행할 수 있을 것으로 사료된다.

• IMMUNE NETWORK
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• 제16권3호
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• pp.183-188
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• 2016

Sirtuin family members with lysine deacetylase activity are known to play an important role in anti-aging and longevity. Cellular senescence is one of the hallmarks of aging, and downregulation of sirtuin is reported to induce premature senescence. In this study, we investigated the effects of small-molecule sirtuin inhibitors on cellular senescence. Various small molecules such as tenovin-1 and EX527 were employed for direct sirtuin activity inhibition. U251, SNB-75, and U87MG glioblastoma cells treated with sirtuin inhibitors exhibited phenotypes with nuclear enlargement. Furthermore, treatment of rat primary astrocytes with tenovin-1 also increased the size of the nucleus. The activity of senescence-associated β-galactosidase, a marker of cellular senescence, was induced by tenovin-1 and EX527 treatment in U87MG glioblastoma cells. Consistent with the senescent phenotype, treatment with tenovin-1 increased p53 expression in U87MG cells. This study demonstrated the senescence-inducing effect of sirtuin inhibitors, which are potentially useful tools for senescence research.

• Journal of Korean Neurosurgical Society
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• 제61권4호
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• pp.441-449
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• 2018

Objective : Notch receptors are heterodimeric transmembrane proteins that regulate cell fate, such as differentiation, proliferation, and apoptosis. Dysregulated Notch pathway signaling has been observed in glioblastomas, as well as in other human malignancies. Nerve growth factor (NGF) is essential for cell growth and differentiation in the nervous system. Recent reports suggest that NGF stimulates glioblastoma proliferation. However, the relationship between NGF and Notch1 in glioblastomas remains unknown. Therefore, we investigated expression of Notch1 in a glioblastoma cell line (U87-MG), and examined the relationship between NGF and Notch1 signaling. Methods : We evaluated expression of Notch1 in human glioblastomas and normal brain tissues by immunohistochemical staining. The effect of NGF on glioblastoma cell line (U87-MG) was evaluated by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. To evaluate the relationship between NGF and Notch1 signaling, Notch1 and Hes1 expression were evaluated by reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis, respectively. To confirm the effects of NGF on Notch1 signaling, Notch1 and Hes1 small interfering RNAs (siRNAs) were used. Results : In immunohistochemistry, Notch1 expression was higher in glioblastoma than in normal brain tissue. MTT assay showed that NGF stimulates U87-MG cells in a dose-dependent manner. RT-PCR and Western blot analysis demonstrated that Notch1 and Hes1 expression were increased by NGF in a dose-dependent manner. After transfection with Notch1 and Hes1 siRNAs, there was no significant difference between controls and 100 nM $NGF-{\beta}$, which means that U87-MG cell proliferation was suppressed by Notch1 and Hes1 siRNAs. Conclusion : These results indicate that NGF stimulates glioblastoma cell proliferation via Notch1 signaling through Hes 1.

• 한국동물위생학회지
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• 제42권4호
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• pp.169-175
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• 2019

Naegleria fowleri, a pathogenic free-living amoeba, leads to a fatal infection known as primary amebic meningoencephalitis (PAM) in human and animals. PAM is an acute, fulminant, necrotizing, and hemorrhagic disease that leads to death in approximately seven days. In this study, we investigate the cytotoxicity of target cells and the secreted molecules of N. fowleri under the non-contact condition. The target cell (U87MG cell) treated with N. fowleri lysates showed no morphological changes and no cytotoxicity. By contrast, the U87MG cells co-cultured with N. fowleri trophozoites under the non-contact condition induced morphological changes and reduction in number. When U87MG cells were co-cultured with N. fowleri trophozoites under the non-contact condition for 30 min, 2 hr, and 4 hr, the levels of cytotoxicity of target cells were 32.3, 35.5, and 37.8%, respectively. Particularly, when the ratio of amoeba to target cells is 10 to 1, the level of cytotoxicity of target cells was 49.7% at 30 min. To show the proteins secreted from N. fowleri under the non-contact condition, we carried out 2D electrophoresis and observed 6 major proteins. Finally, these results suggest that the molecules released from N. fowleri under the non-contact condition induce the cell death and this process is an important step in pathogenesis of N. fowleri.

• 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.

• 대한임상검사과학회지
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• 제51권2호
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• pp.145-154
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• 2019

본 연구는 대한민국 성인에서 eGFR 및 uACR과 Ferritin의 관련성에 대한 연구이다. 2012년 국민건강영양조사자료에서 20세 이상의 4,948명을 대상으로 관련변수를 보정한 후, 만성신장질환(CKD, eGFR<$60mL/min/1.73m^2$) 및 알부민뇨($uACR{\geq}30mg/g$)에 따른 페리틴 수준을 분석하였다. 만성신장질환군의 ferritin 수준($M{\pm}SE$) [$103.04{\pm}6.59mL/min/1.73m^2$; 95% confidence interval (CI), 90.12~115.96]은 정상군($84.87{\pm}1.16mL/min/1.73m^2$; 95% CI, 82.59~87.14)에 비하여 유의하게 높았다(P=0.007). 그러나 정상군($82.72{\pm}4.09mg/g$; 95% CI, 74.71~90.73)과 알부빈뇨군($82.72{\pm}4.09mg/g$; 95% CI, 74.71~90.73)의 ferritin 수준은 유의한 차이가 없었다(P=0.487). 결과적으로, 대한민국 성인에서 만성신장질환과 ferritin수준은 양의 상관관계가 있었지만, 알부빈뇨에서는 유의한 차이가 없었다.

• Asian Pacific Journal of Cancer Prevention
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• 제14권4호
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• pp.2607-2610
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• 2013

Malignant glioma, also known as brain cancer, is the most common intracranial tumor, having an extremely high mortality and recurrence rate. The survival rate of the affected patients is very low and treatment is difficult. Hence, growth inhibition of glioma has become a hot topic in the study of brain cancer treatment. Among the various isothiocyanate compounds, it has been confirmed that benzyl isothiocyanate (BITC) can inhibit the growth of a variety of tumors, including leukemia, glioma and lung cancer, both inside and outside the body. This study explored inhibitory effects of BITC on human glioma U87MG cells, as well as potential mechanisms. It was found that BITC could inhibit proliferation, induce apoptosis and arrest cell cycling of U87MG cells. In addition, it inhibited the expression of SOD and GSH, and caused oxidative stress to tumor cells. Therefore, it is believed that BITC can inhibit the growth of U87MG cells outside the body. Its mechanism may be related to the fact that BITC can cause oxidative stress to tumor cells.

• 대한방사성의약품학회지
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• 제5권2호
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• pp.101-112
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• 2019

Herein, 2-nitroimidazole-fluorophore conjugates were synthesized by linking 2-nitroimidazole and FITC or RITC via thiourea bonds. The prepared derivatives were stable for 2 h in Dulbecco's modified Eagle's medium (DMEM) at 37 ℃. The novel conjugates were studied for their in vitro uptake under hypoxic conditions using U87MG and CT-26 cell lines, showing significantly higher uptakes in hypoxic than normoxic cells. Immunohistochemical analysis confirmed hypoxia in U87MG and CT-26 xenografted tumor tissues. Moreover, the prepared conjugates were evaluated by in vivo experiments after intravenous injection in U87MG and CT-26 xenografted mice. Hypoxia was confirmed by immunohistochemistry of the prepared derivatives with co-injected pimonidazole. Confocal microscopy of the prepared derivatives showed strong fluorescence in hypoxic tumor tissues correlated with the pimonidazole distribution. This suggested that the 2-nitroimidazole-fluorophore conjugates are promising optical imaging probes for tumor hypoxia and are promising substitutes for pimonidazole immunohistochemistry, which requires a multi-step procedure of incubation involving antibody, second antibody, dye, hydrogen peroxide, and multiple washing steps.

• Journal of Korean Neurosurgical Society
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• 제29권4호
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• pp.471-476
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• 2000

Objective : p16/INK4a, a kind of tumor suppressor genes, encodes a specific inhibitor of the cyclin D-dependent kinases CDK4 and CDK6. This prevents the association of CDK4 with cyclin D1, and subsequently inhibits phosphorylation of retinoblastoma tumor suppressor protein(pRb), thus preventing exit from the G1 phase. According to previous reports, over 50% of glioma tissue and 80% of glioma cell lines have been demonstrated inactivation of p16/INK4a gene. The purpose of this study was to determine whether recombinant adenovirus-p16 virus is a suitable candidate for gene replacement therapy in cases of glioma. Methods : Three human glioma cell lines(U251MG, U87MG and U373MG) that express mutant p16 protein were used. Replication-deficient adenovirus was utilized as an expression vector to transfer exogenous p16 cDNA into the cells ; control cells were infected with the Ad-${\beta}$-gal expressing ${\beta}$-galactosidase. To monitor gene transfer and the expression of exogenous genes, we used Western Blotting analysis. Flow cytometry studies of cellular DNA content were performed to determine the cell cycle phenotype of the glioma cells before and after treatment. Results : We showed here that restoration of p16/INK4a expression in p16 negative U87MG, U251MG and partially deleted U373MG by Ad-CMV-p16 induced growth suppression in vitro. Flow cytometric study revealed that Ad-CMV-p16 infected U87MG cells were arrested during the G0-G1 phase of the cell cycle. Expression of p16 transferred by Ad-CMV-p16 in glioma cells was highly efficient and maintained for more than seven days. Conclusions : Our results suggest that Ad-CMV-p16 gene therapy strategy is potentially useful and warrants further clinical investigation for the treatment of gliomas.

• Journal of Korean Neurosurgical Society
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• 제55권3호
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• pp.131-135
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• 2014

Objective : With the growing interests of bacteria as a targeting vector for cancer treatment, diverse genetically engineered Salmonella has been reported to be capable of targeting primary or metastatic tumor regions after intravenous injection into mouse tumor models. The purpose of this study was to investigate the capability of the genetically engineered Salmonella typhimurium (S. typhimurium) to access the glioma xenograft, which was monitored in mouse brain tumor models using optical bioluminescence imaging technique. Methods : U87 malignant glioma cells (U87-MG) stably transfected with firefly luciferase (Fluc) were implanted into BALB/cAnN nude mice by stereotactic injection into the striatum. After tumor formation, attenuated S. typhimurium expressing bacterial luciferase (Lux) was injected into the tail vein. Bioluminescence signals from transfected cells or bacteria were monitored using a cooled charge-coupled device camera to identify the tumor location or to trace the bacterial migration. Immunofluorescence staining was also performed in frozen sections of mouse glioma xenograft. Results : The injected S. typhimurium exclusively localized in the glioma xenograft region of U87-MG-bearing mouse. Immunofluorescence staining also demonstrated the accumulation of S. typhimurium in the brain tumors. Conclusion : The present study demonstrated that S. typhimurium can target glioma xenograft, and may provide a potentially therapeutic probe for glioma.