• Title/Summary/Keyword: 암 세포주

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Analysis of Efficiency of Recombinant pOPINEneo-3C-GFP Vector with p53 Tumor Suppression Gene Inserted (p53 암억제 유전자가 삽입된 재조합 pOPINEneo-3C-GFP 벡터의 효율 분석)

  • Sa, Young-Hee;Choi, Chang-Shik;Lee, Ki Hwan;Hong, Seong-Karp
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 2019.05a
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    • pp.533-536
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    • 2019
  • Recombinant baculoviruses are widely used to express heterologous genes in cultured insect cells. Recombinant baculoviruses can serve as gene-transfer vectors for expression of recombinant proteins in a wide range of mammalian cell types. Baculovirus system has significant benefits in view of safety, large-scale, and high level of gene expression. In this study, baculoviral vectors which were reconstructed from pOPINEneo-3C-GFP vector, were recombined with cytomegalovirus (CMV) promoter, green fluorescent protein (GFP), and p53 with NcoI and XhoI. These recombinant vectors were infected with various cells and cell lines. The baculovirus vector thus developed was analyzed by comparing the metastasis and expression of the recombinant genes with conventional vectors. These results suggest that the baculovirus vector has higher efficiency in metastasis and expression than the control vector. This work was supported by a grant from Mid-Career Researcher Program(NRF-2016R1A2B4016552) through the National Research Foundation of Korea(NRF) funded by the Ministry of Science, ICT & Future Planning(MSIP).

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Correlation of Protumor Effects of Leucine-Rich Repeat Kinase 2 with Interleukin-10 Expression in Lung Squamous Cell Carcinoma (폐 편평세포암종 내 Leucine-Rich Repeat Kinase 2 암촉진 효과와 Interleukin-10 발현과의 연관성)

  • Sung Won LEE;Sangwook PARK
    • Korean Journal of Clinical Laboratory Science
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    • v.55 no.2
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    • pp.105-112
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    • 2023
  • Leucine-rich repeat kinase 2 (LRRK2) is known to play a crucial role in the pathophysiology of neurodegenerative disorders such as Parkinson's disease. LRRK2 is predominantly expressed in the lung as well as the brain. However, it is unclear whether LRRK2 expression correlates with the pathogenesis of lung squamous cell carcinoma (LUSC). This study analyzes the prognostic significance of LRRK2 in LUSC using the Kaplan-Meier plotter tool. High expression of LRRK2 is known to be associated with a bad prognosis in patients with LUSC. Patients with high LRRK2 expression, tumor mutational burden, high neoantigen load, and even gender correlation reportedly have the worse survival rates. In the gene expression profiling interactive analysis (GEPIA) database, the severity of pathogenesis in LUSC with high LRRK2 expression positively corresponds to a high expression of anti-inflammatory cytokines but not inflammatory cytokines. Similarly, the increased expression of interleukin (IL)10-related genes was shown to be significantly linked in LRRK2-high LUSC patients having a poor prognosis. Moreover, the tumor immune estimation resource (TIMER) database suggests that macrophages are one of the cellular sources of IL10 in LRRK2-high LUSC patients. Collectively, our results demonstrate that the postulated LRRK2-IL10 axis is a potential therapeutic target and prognostic biomarker for LUSC.

Factors Influencing the Activation of Brown Adipose Tissue in 18F-FDG PET/CT in National Cancer Center (양전자방출단층촬영 시 갈색지방조직 활성화에 영향을 미치는 요인 분석)

  • You, Yeon Wook;Lee, Chung Wun;Jung, Jae Hoon;Kim, Yun Cheol;Lee, Dong Eun;Park, So Hyeon;Kim, Tae-Sung
    • The Korean Journal of Nuclear Medicine Technology
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    • v.25 no.1
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    • pp.21-28
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    • 2021
  • Purpose Brown fat, or brown adipose tissue (BAT), is involved in non-shivering thermogenesis and creates heat through glucose metabolism. BAT activation occurs stochastically by internal factors such as age, sex, and body mass index (BMI) and external factors such as temperature and environment. In this study, as a retrospective, electronic medical record (EMR) observation study, statistical analysis is conducted to confirm BAT activation and various factors. Materials and Methods From January 2018 to December 2019, EMR of patients who underwent PET/CT scan at the National Cancer Center for two years were collected, a total of 9155 patients were extracted, and 13442 case data including duplicate scan were targeted. After performing a univariable logistic regression analysis to determine whether BAT activation is affected by the environment (outdoor temperature) and the patient's condition (BMI, cancer type, sex, and age), A multivariable regression model that affects BAT activation was finally analyzed by selecting univariable factors with P<0.1. Results BAT activation occurred in 93 cases (0.7%). According to the results of univariable logistic regression analysis, the likelihood of BAT activation was increased in patients under 50 years old (P<0.001), in females (P<0.001), in lower outdoor temperature below 14.5℃ (P<0.001), in lower BMI (P<0.001) and in patients who had a injection before 12:30 PM (P<0.001). It decreased in higher BMI (P<0.001) and in patients diagnosed with lung cancer (P<0.05) In multivariable results, BAT activation was significantly increased in patients under 50 years (P<0.001), in females (P<0.001) and in lower outdoor temperature below 14.5℃ (P<0.001). It was significantly decreased in higher BMI (P<0.05). Conclusion A retrospective study of factors affecting BAT activation in patients who underwent PET/CT scan for 2 years at the National Cancer Center was conducted. The results confirmed that BAT was significantly activated in normal-weight women under 50 years old who underwent PET/CT scan in weather with an outdoor temperature of less than 14.5℃. Based on this result, the patient applied to the factor can be identified in advance, and it is thought that it will help to reduce BAT activation through several studies in the future.

Plant regeneration from suspension-cultured cell clusters of Arabidopsis thaliana (애기장대(Arabidopsis thaliana)의 현탁배양세포괴로부터 식물체 재분화)

  • 김명덕;김준철;진창덕;임창진;한태진
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.3
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    • pp.195-200
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    • 1998
  • Callus induction from leaf and stem explants of Arabidopsis thaliana was successfully obtained when leaf explants were cultured on MS medium containing 2.0 mg/L 2,4-D in the dark and also, when stem explants were cultured on CP medium containing 0.5 mg/L 2,4-D and 0.1 mg/L BAP. Explant-derived sliced calli were suspension-subcultured every week in CP liquid medium with 0.5 mg/L 2,4-D and 0.1 mg/L BAP in the dark, and shoot-forming cell clusters of nodular, pale yellow and knobby type were selected after 7-8 weeks of culture. Shoots were initiated from the green spots of the selected shoot forming calli cultured on MS regeneration medium containing 0.05 mg/L IAA, 7.0 mg/L 2-iP and 30 g/L sucrose under continous illumination for four weeks. Shoot regeneration frequency (calli regenerating at least one shoot) was more than 50%. For plant regeneration, excised shoots were trnasferred to hormone free medium for root initiation after 4 weeks of culture. The regenerants were bolting after 2 weeks of culture and formed in vitro flowering buds within bracts after 4 weeks of culture.

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Concurrent Chemoradiation in Patients with Cancer of the Esophagus (식도암에서의 동시화학방사선요법)

  • Lee Kang Kyoo;Park Kyung Ran;Lee Jong Young;Shin Hyun Soo;Lee Chong In;Chang Woo Ick;Shim Young Hak
    • Radiation Oncology Journal
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    • v.16 no.1
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    • pp.7-16
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    • 1998
  • Purpose : To evaluate survival rate and prognostic factors affecting survival of patients with esophageal cancer treated with concurrent chemoradiation. Materials and Methods : Eligibility included biopsy proven invasive carcinoma of the cervical or thoracic esophagus, confined to esophagus and mediastinum with or without regional lymph node and supraclavicular lymph node, and ECOG Performance status $H_0-H_2$. Patients received radiation therapy with 5940cGy over 7 weeks and chemotherapy, consisted of 5-FU(1000 $mg/m^2/day$ in continuous infusion for 5 days, days 1 to 5 and days 29 to 33) and mitomycin C($8mg/m^2$ intravenous bolus at day 1). After concurrent chemoradiation, maintenance chemotherapy was followed with 5-FU(1000 $mg/m^2/day$ in continuous infusion for 5 days at 9th, 13th, and 17th weeks) and cisplatin($80mg/m^2$ intravenous bolus at the first day of each cycle). Results : From November 1989 to November 1995, 44 patients were entered in this study. After treatment, complete response rate and partial response rate were $59\%$ and $41\%$. Overall 1, 2, and 5-year survivals were $59\%$, $38\%$, and $9.6\%$(median 17 months), Prognostic factors affecting survival were response to treatment and T-stage. Among 26 complete responders, there were 6 local recurrences, 3 distant recurrences, 1 local and distant recurrence, and 2 unknown site recurrences Acute and chronic complication rates with grade 3 or more were $20\%$ and $13.0\%$ and there was no treatment-related mortality. Conclusion : Concurrent chemoradiation, compared with historical control groups that treated with radiation alone, improved median survival and did not significantly increase treatment-related complications. Complete responders had longer survival duration than partial responders. Predominant failure pattern was local failure. So, efforts to improve local control should be proposed.

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Phytic Acid Protects the Formation of Colonic Aberrant Crypt Foci Induced by Azoxymethane in Male F344 Rats (랫드에서 azoxymethane으로 유도된 대장 전암병변에 대한 피티산의 방어 효과)

  • Hue, Jin-Joo;Lee, Yea-Eun;Lee, Ki-Nam;Nam, Sang-Yoon;Ahn, Byeong-Woo;Yun, Young-Won;Lee, Beom-Jun
    • Journal of Food Hygiene and Safety
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    • v.23 no.3
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    • pp.264-270
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    • 2008
  • Phytic acid(PA) (Inositol hexaphosphate, $IP_6$) is a naturally occurring polyphosphorylated carbohydrate that is present in substantial amounts in almost all plants and mammalian cells. Recently PA has received much attention for its role in anticancer activity. In the present study, the preventive effects of PA on colon carcinogenesis were investigated. Six-week old Fisher 344 male rats were fed a AIN-93G purified diet and PA(0.5% or 2% PA in water) for 8 weeks. The animals received two ($1^{st}\;and\;2^{nd}$ week) injections of azoxymethane(AOM, 15 mg/kg b.w.) to induce colonic aberrant crypt foci(ACF). After sacrifice, the total numbers of aberrant crypts(AC) and ACF in colonic mucosa were examined after staining with methylene blue. Blood and serum were analyzed with a blood cell differential counter and an automatic serum analyzer. AOM induced the total numbers of $142.3{\pm}22.3$ ACF/colon and $336.6{\pm}55.1$ AC/colon. PA at the doses of 0.5 and 2% decreased the numbers of ACF and AC/colon in a dose-dependent manner. The numbers of ACF/colon and AC/colon by PA at the dose of 0.5% were $124.4{\pm}28.5\;and\;302.7{\pm}67.3$, respectively. PA at the dose of 2% significantly decreased the ACF and AC numbers to $109{\pm}18.1\;and\;254.8{\pm}50.6$, respectively(p<0.01). Especially, 2% PA significantly reduced the number of large ACF(${\geq}4$ AC/ACF) from $26.8{\pm}6.2$ ACF/colon to $15{\pm}6.7$ ACF/colon(p<0.01). Although some parameters in blood counts and serum chemistry were changed compared with the control, no specific toxicity was found. These findings suggest that phytic acid can be a chemopreventive agent for colon carcinogenesis resulting from inhibition of the development of ACF in the F344 rat.

In Vitro Study of Tumor Seeking Radiopharmaceutical Uptake by Human Breast Cancer Cell Line MCF-7 after Paclitaxel Treatment (사람 유방암세포주 MCF-7에 Paclitaxel 처치 후 종양영상용 방사성의약품 섭취 변화에 대한 시험관내 연구)

  • Choi, Joon-Young;Choi, Yong;Choe, Yearn-Seong;Lee, Kyung-Han;Kim, Byung-Tae
    • Nuclear Medicine and Molecular Imaging
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    • v.41 no.5
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    • pp.364-372
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    • 2007
  • Purpose: This study was designed to investigate the cellular uptake of various tumor imaging radiopharmaceuticals in human breast cancer cells before and after paclitaxel exposure considering viable cell number. Materials and Methods: F-18-fluorodeoxyglucose, C-11-methionine, Tl-201, Tc-99m-MIBI, and Tc-99m-tetrofosmin were used to evaluate the cellular uptake in MCF-7 cells. MCF-7 cells were cultured in multi-well plates. Wells were divided into DMSO exposure control group, and paclitaxel exposure group. The exposure durations of paclitaxel with 10 nM or 100 nM were 2 h, 6 h, 12 h, 24 h, and 48 h. Results: Viable cell fraction was reduced as the concentration and exposure time of paclitaxel increased. After 10 nM paclitaxel exposure, the cellular uptake of all 5 radiopharmaceuticals was not reduced significantly, irrespective of exposure time and viable cell fraction. After 100 nM paclitaxel exposure, the cellular uptake of all 5 radiopharmaceuticals was enhanced significantly irrespective of viable cell fraction. The peak uptake was observed in experimental groups with paclitaxel exposure for 6 to 48 h according the type of radiopharmaceutical. When the cellular uptake was adjusted for the viable cell fraction and cell count, the peak cellular uptake was observed in experimental groups with paclitaxel exposure for 48 h, irrespective of the type of radiopharmaceutical. Conclusion: The cellular uptake of F-18-fluorodeoxyglucose, C-11-methionine, Tl-201, Tc-99m-MIBI, and Tc-99m-tetrofosmin did not reflect viable cell number in MCF-7 cells after paclitaxel exposure for up to 48 h.

Conditioned Media of RAW 264.7 Cells Stimulated with Phellinus linteus Extract Regulates the Epithelial-mesenchymal Transition in Prostate Cancer Cells (상황버섯에 의해 활성화된 RAW 264.7 대식세포주 배양액의 인간 전립선암 세포주의 epithelial-mesenchymal transition 조절)

  • Kang, Taewoo;An, Hyun-Hee;Park, Sul-Gi;Yu, Sun-Nyoung;Hwang, You-Lim;Kim, Ji-Won;Ahn, Soon-Cheol
    • Journal of Life Science
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    • v.29 no.8
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    • pp.904-915
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    • 2019
  • Prostate cancer (PCa) is one of the most metastatic tumor. Although hormone therapy or surgical castration is mostly conducted to treat PCa, it has a lot of side effects. Recently, many researchers have been exploring the tumor microenvironment to remedy these circumstances. Immune cells, especially macrophages, are an important composition of the tumor microenvironment. Under normal conditions, macrophages exhibit mild tumoricidal activity against tumors. However, once activated by interferon gamma or lipopolysaccharides, macrophages can kill cancer cells directly or indirectly by secreting cytokines and chemokines. In this study, murine macrophage RAW 264.7 cells were treated with Phellinus linteus extract. To analyze their pro-inflammatory phenotype, we were used several assays such as a real-time polymerase chain reaction, an enzyme-linked immunosorbent and nitric oxide assay. Prostate cancer cells were treated with the RAW 264.7-conditioned media, which was identified as a pro-inflammatory nature, for 48 h, and the expression of epithelial-mesenchymal transition (EMT)-related genes was determined. Not only N-cadherin, Snail, Twist, Slug, and Cadherin 11, which are mechenchymal-related proteins, were decrease, but epithelial marker of E-cadherin was increased. In addition, the mRNA level of vimentin, ccl2, and vegfa were decreased, as the EMT is closely related to the migration and invasion of cancer cells. In conclusion, the RAW 264.7-conditioned media stimulated with P. linteus extract inhibited migration and invasion and regulated the EMT pathway in human prostate cancer cells.

AMP-activated Kinase Regulates Adipocyte Differentiation Process in 3T3-L1 Adipocytes Treated with Selenium (AMP-activated protein kinase가 셀레늄으로 처리된 3T3-L1 지방세포의 분화과정 억제에 관한 연구)

  • Park, Song-Yi;Hwang, Jin-Taek;Lee, Yun-Kyoung;Kim, Young-Min;Park, Ock-Jin
    • Journal of Life Science
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    • v.19 no.4
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    • pp.423-428
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    • 2009
  • Selenium was investigated using human origin preadipocytes to see whether it possesses preventive or therapeutic effects for obesity. Unveiling the potential of selenium in the reduction of adipogenesis can help predict the therapeutic capabilities of selenium in obesity. In the present study, the molecular mechanism of the inhibition of adipogenesis by selenium was explored to unravel the involvement of the AMP-activated protein kinase. There is emerging evidence that AMPK, a sensor of cellular energy status, is a possible molecular target of controlling adipocyte differentiation on the basis of discovery that AMPK is responsible for the major metabolic responses to exercise, and integration of nutritional and hormonal signals to modulate feeding behavior or energy expenditure in the hypothalamus. Treatment of selenium resulted in inhibition of the adipocyte differentiation process and induction of mature apoptosis in 3T3-L1 adipocytes. We hypothesized that selenium may exert anti-adipogenic potential though modulating AMPK. We have found that selenium significantly activated AMPK and phosphorylated its substrate acetyl-CoA carboxylase ($ACC-serine^{79}$) during the inhibitory process of adipocytes. Also, the inhibition process of adipocyte differentiation by selenium was comparable to either reveratrol or a synthetic AMPK activator, AICAR (5-aminoimidazole-4-carboxamide-1-${\beta}$-D-ribofuranoside). To evaluate the involvement of AMPK in anti-lipogensis, we applied AICAR and Compound C, an AMPK inhibitor, to 3T3-L1-adipocytes and found that AMPK is required for the adipocyte differentiation blocking process. These results suggest that selenium has a potential to control adipogenesis and that this effect is mediated by AMPK, an essential kinase for both inhibition of adipocyte differentiation and apoptosis of mature adipocytes.

Comparison of the Uptakes of $^{99m}Tc-sestamibi\;and\;^{99m}Tc-tetrofosmin$ in Cancer Cell Lines Expressing Multidrug Resistance (다약제내성 발현 암세포에서 $^{99m}Tc-sestamibi$$^{99m}Tc-tetrofosmin$ 섭취의 비교)

  • Yoo, Jeong-Ah;Chung, Shin-Young;Seo, Myeng-Rang;Kwak, Dong-Suk;Ahn, Byeong-Cheol;Lee, Kyu-Bo;Lee, Jae-Tae
    • The Korean Journal of Nuclear Medicine
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    • v.37 no.3
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    • pp.178-189
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    • 2003
  • Purpose: Cellular uptakes of $^{99m}Tc-sestamibi(MIBI)\;and\;\;^{99m}Tc-tetrofosmin$ into cancer cell lines expressing multidrug resistance(MDR) were investigated and compared. The effects of verapamil and cyclosporin A, well-known multidrug resistant reversing agents, on cellular uptakes of both tracers were also compared. Materials and Methods: Doxorubicin-resistant HCT15/CL02 human colorectal cell and doxorubicin-resistant K562(Adr) and vincristine-resistant K562(Vcr) human leukemic cells were studied. RT-PCR analysis was used for the detection of mdr1 mRNA expression. MDR-reversal effects with verapamil and cyclosporine A were evaluated at different drug concentrations after incubation with MIBI and tetrofosmin for 1, 15, 30, 45 and 60 min, using single-cell suspensions at $1{\times}10^6cells/ml$ incubated at $37^{\circ}C$. Radioactivity in supernatants and pellets were measured with gamma well counter. Results: The cellular uptakes of MIBI and tetrofosmin in K562(Adr) and K562(Vcr) were lower than those of parental K562 cell. In HCT15/CL02 cells and K562(Adr) cells, there were no significant difference in cellular uptakes of both tracers, but cellular uptake of MIBI was higher than that of tetrofosmin in K562(Vcr) cells. Coincubation with verapamil resulted in a increase In cellular uptakes of MIBI and tetrofosmin. Verapamil increased cellular uptakes of MIBI and tetrofosmin by HCT15/CL02 cell by 11.9- and 6.8-fold, by K562(Adr) cell by 14.3- and 8-fold and by K562(Vcr) cell by 7- and 5.7-fold in maximum, respectively. Cyciosporin A increased cellular uptakes of MIBI and tetrofosmin by HCT15/CL02 cell by 10- and 2.4-fold, by K562(Adr) cell by 44- and 13-fold and by K562(Vcr) cell by 18.8- and 11.8-fold in maximum, respectively Conclusion: Taking together, MIBI and tetrofosmin are considered as suitable radiopharmaceuticals for defecting multidrug resistance. However, MIBI seems to be a better tracer than tetrofosmin for evaluating MDR reversal effect of the modulators. Since cellular uptakes of both tracers might differ in different cell types, further experiments regarding differences in cellular uptakes between cell types should be explored.