• Title/Summary/Keyword: CT-26 cell

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Apoptotic Pathway Induced by Dominant Negative ATM Gene in CT-26 Colon Cancer Cells (CT-26 대장암 세포에서 Dominant Negative ATM 유전자에 의하여 유도되는 세포자멸사의 경로)

  • Lee, Jung Chang;Yi, Ho Keun;Kim, Sun Young;Lee, Dae Yeol;Hwang, Pyoung Han;Park, Jin Woo
    • Clinical and Experimental Pediatrics
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    • v.46 no.7
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    • pp.679-686
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    • 2003
  • Purpose : Ataxia telangiectasia mutated(ATM) is involved in DNA damage responses at different cell cycle checkpoints, and signalling pathways associated with regulation of apoptosis in response to ionizing radiation(IR). However, the signaling pathway that underlies IR-induced apoptosis in ATM cells has remained unknown. The purpose of this study was, therefore, to investigate the apoptotic pathway that underlies IR-induced apoptosis in a CT-26 cells expressing dominant negative ATM (DN-ATM). Methods : We generated a replication-deficient recombinant adenovirus encoding the DN-ATM(Ad/DN-ATM) or control adenovirus encoding no transgene(Ad/GFP) and infected adenovirus to CT-26 cells. After infection, we examined apoptosis and apoptotic pathway by [$^3H$]-thymidine assay, DNA fragmentation, and Western immunoblot analysis. Results : DN-ATM gene served as the creation of AT phenotype in a CT-26 cells as revealed by decreased cell proliferations following IR. In addition, IR-induced apoptosis was regulated through the reduced levels of the anti-apoptotic protein Bcl-2, the increased levels of the apoptotic protein Bax, and the activation of caspase-9, caspase-3, and PARP. Conclusion : These results indicate that the pathway of IR-induced apoptosis in CT-26 cells expressing DN-ATM is mediated by mitochondrial signaling pathway involving the activation of caspase 9, caspase 3, and PARP.

Exosomes from CIITA-Transfected CT26 Cells Enhance Anti-tumor Effects

  • Fan, Wen;Tian, Xing-De;Huang, E.;Zhang, Jia-Jun
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.2
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    • pp.987-991
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    • 2013
  • Aim: To study anti-tumor effects of exosomes from class II transactivator (CIITA) gene transfected CT26 cells. Methods: In this study, we established an MHC class II molecule-expressing murine colon cancer cell line (CT26-CIITA) by transduction of the CIITA gene. Immune effects in vitro and tumor protective results in vivo were tested and monitored. Results: Exosomes from CT26-CIITA cells were found to contain a high level of MHC class II protein. When loaded on dendritic cells (DCs), exosomes from CT26-CIITA cells significantly increased expression of MHC class II molecules, CD86 and CD80, as compared to exosomes from CT26 cells. In vitro assays using co-culture of immunized splenocytes and exosome-loaded DCs demonstrated that CIITA-Exo enhanced splenocyte proliferation and IFN-${\gamma}$ production of CD4+T cells, while inhibiting IL-10 secretion. In addition, compared to exosomes from CT26 cells, CT26-CIITA-derived exosomes induced higher TNF-${\alpha}$ and IL-12 mRNA levels. A mouse tumour preventive model showed that CT26-CIITA derived exosomes significantly inhibited tumour growth in a dose-dependent manner and significantly prolonged the survival time of tumour-bearing mice. Conclusion: Our findings indicate that CT26-CIITA-released exosomes are more efficient to induce anti-tumour immune responses, suggesting a potential role of MHC class II-containing tumour exosomes as cancer vaccine candidates.

The Use of MTT Assay, In Vitro and Ex Vivo, to Predict the Radiosensitivity of Colorectal Cancer (In-vitro와 Ex-vivo MTT Assay를 통한 직장암의 방사선치료 감수성 예측 가능성 검증)

  • Kim, Ji-Eun;Kim, Mi-Sook;Kang, Chang-Mo;Kim, Jong-Il;Shin, Hye-Kyung;Choi, Chul-Won;Seo, Young-Seok;Ji, Young-Hoon
    • Radiation Oncology Journal
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    • v.26 no.3
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    • pp.166-172
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    • 2008
  • Purpose: The measurement of radiosensitivity of individuals is useful in radiation therapy. Unfortunately, the measurement of radiation survival using a clonogenic assay, which is the established standard, can be difficult and time consuming. The aim of this study is to compare radiosensitivity results obtained from the MTT and clonogenic assays, and to evaluate whether the MTT assay can be used on clinical specimens. Materials and Methods: HCT-8, LoVo, CT-26, and WiDr were the colon cancer cell lines used for this study. The clonogenic assay was performed to obtain the cell survival curves and surviving fractions at a dose of 2 Gy ($SF_2$) as the standard technique for radiosensitivity. Also, the MTT assay was performed for each of the cell lines (in vitro). To simulate clinical specimens, the cell lines were inoculated into nude mice, removed when the tumors reached 1 cm in diameter, and chopped. Next, the tumors were subjected to the same process involved with the MTT assay in vitro. The inhibition rates (IR) of 10 Gy or 20 Gy of irradiation for in vitro and ex vivo were calculated based on the optical density of the MTT assay, respectively. Results: According to $SF_2$ and the cell survival curve, the HCT-8 and WiDr cell lines were more resistant to radiation than LoVo and CT-26 (p<0.05). The IR was measured by in vitro. The MTT assay IR was 17.3%, 21%, 30% and 56.5% for the WiDr, HCT-8, LoVo and CT-26 cell lines, respectively. In addition, the IR measured ex vivo by the MTT assay was 23.5%, 26%, 38% and 53% in the HCT-8, WiDr, LoVo and CT-26 tumors, respectively. Conclusion: The radiosensitivity measured by the MTT assay was correlated with the measures obtained from the clonogenic assay. This result highlights the possibility that the MTT assay could be used in clinical specimens for individual radiosensitivity assays.

Anti-Oxidative and Anti-Inflammatory Effects of Cheongajihwang-Tang Extract on RAW264.7 Cells (청아지황탕(靑蛾地黃湯)의 RAW264.7 Cell에서 항산화 및 항염증 효능 연구)

  • Yoon, Jeong-won;Kim, Soon-Joong;Park, Dong-Su
    • Journal of Korean Medicine Rehabilitation
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    • v.26 no.3
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    • pp.51-58
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    • 2016
  • Objectives This study was designed to investigate whether the Cheongajihwang-Tang (CT) has an inhibitory effect association with oxidation or inflammation in RAW264.7 cells. Methods Cytotoxic activity of CT extract on RAW264.7 cells was evaluated by using 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide (MTT) solution. Nitric oxide production was measured using Griess reagent system. The total phenolic contents and 1,1-Diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity was measured to evaluate the anti-oxidative effects of CT. Dichlorofluorescin diacetate (DCFH-DA) has been used as a substrate for measuring intracellular oxidant production. Results Cheongajihwang-Tang does not impair the cell viability in tested concentration. CT showed anti-oxidative effects in vitro by decreasing electron donating ability, and also showed anti-inflammatory effects suppressing NO and ROS expression in LPS induced RAW264.7 activation. CT inhibited the generation of intracellular ROS production as dose dependant manner. Conclusions CT has anti-oxidative effects and anti-inflammatory activities. These results indicate that CT extract has an anti-inflammatory activities via anti-oxidative effects.

Computer Tomography and Magnetic Resonance Image Manifestations of Primary Hepatic Neuroendocrine Cell Carcinomas

  • Huang, Juan;Yu, Jian-Qun;Sun, Jia-Yu
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.6
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    • pp.2759-2764
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    • 2014
  • Aim: This study aims to investigate the manifestation of CT, MRI and dynamic enhanced scans for primary hepatic neuroendocrine cell carcinoma. Methods: CT or MRI arterial and venous phase scan images of 19 cases of pathologically confirmed PHNEC were retrospectively analyzed. Results: 14 cases (73.68%) with single lesion, 5 cases (26.3%) with multiple lesions, with an average diameter of 13.2 cm. Some 12 cases (63.16%) showed inhomogeneous enhancement, seven cases (36.8%) showed homogeneous enhancement, 13 cases (68.4%) demonstrated significant enhancement in the arterial phase, 13 cases (68.4%) had significantly enhanced portal venous phase including 7 cases (36.8 %) with portal venous phase density or signal above the arterial phase and 5 cases (26.3%) with the portal vein density or signal below the arterial phase. Seven cases (36.8%) had continued strengthened separate shadows in the center of the lesion. Thrombosis were not seen in portal veins. Conclusion: CT and MRI images of liver cell neuroendocrine carcinoma have certain characteristics that can provide valuable information for diagnosis and differential diagnosis.

Comparison Analysis of Immune Cells between CT26 Tumor Bearing Mice and Normal Mice

  • Lee, Na Kyung;Kim, Hong Sung
    • Biomedical Science Letters
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    • v.20 no.3
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    • pp.147-155
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    • 2014
  • It has well studied that immune cells are strongly related to tumor progression and tumor suppression. To identify the difference of immune cell between tumor bearing mice and normal mice, we examined systemically the immune cell of CT26 tumor bearing mice on 21 days after tumor cell administration. As previously reported, CD4+ and CD8+ T cells population of tumor bearing mice significantly decreased 38% and 30% on day 21 compared to that of normal mice, respectively. All subpopulation of CD4 and CD8+ T cell significantly decreased, except CD49b+ T cell subpopulation. But, myeloid cell population ($CD11b^{high}$ and all Gr-1+ subpopulation) of tumor bearing mice significantly increased on day 21. Especially, all subpopulation of CD11b+Gr-1+ cell of tumor bearing mice significantly increased on day 21. Also, Foxp3+$CD25^{high}$ CD4 T cell (regulatory T cells) population significantly increased on day 21. These results suggest that tumor can induce the decline of T lymphocyte and the expansion of myeloid cells and regulatory T cells, and provide the basic information for the study of tumor immunology.

Usefulness of F-18 FDG PET/CT in Staging of Peripheral T Cell Lymphoma (말초성 T 세포 림프종의 병기 설정시 F-18 FDG PET/CT의 유용성)

  • Kang, Yun-Hee;Lim, Seok-Tae;Kim, Dong-Wook;Jeong, Hwan-Jeong;Sohn, Myung-Hee;Yim, Chang-Yeol
    • Nuclear Medicine and Molecular Imaging
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    • v.42 no.5
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    • pp.369-374
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    • 2008
  • Purpose: F-18 FDG PET/CT has excellent sensitivity and specificity for staging non-Hodgkin lymphomas, but to the author's knowledge few studies to date have evaluated FDG PET/CT in peripheral T cell lymphoma. We evaluated the usefulness of F-18 FDG PET/CT in staging of patients with peripheral T cell lymphoma, especially indolent cutaneous T cell lymphomas. Materials and Methods: Twenty five patients (M:F=17:8, age $53.7{\pm}14.8$ yrs) with biopsy-proven indolent cutaneous T cell (CL) or noncutaneous T cell lymphomas (NCL) underwent PET/CT scans for staging at baseline. Peak standardized uptake values (p-SUV) of all abnormal foci were measured and compared between cutaneous and noncutaneous lesions. F-18 FDG PET/CT was performed on 6 patients with indolent CL and on 19 patients with NCL. Results: All 6 patients with indolent CL had no significant FDG avidity in the skin despite histologically positive cutaneous lesions. However, FDG avidity appeared in extracutaneous lesions (lymph nodes) in two patients with CL where CT imaging suggested lymphoma involvement (mean p-SUV $4.26{\pm}0.37$ in noncutaneous lesions in CL). In NCL, FDG avidity was demonstrated in all lesions where CT imaging suggested lymphoma involvement (mean p-SUV, $8.52{\pm}5.00$ in noncutaneous lesions in NCL). Conclusion: F-18 FDG PET/CT has the limitation of usefulness for the evaluation of the skin in indolent CL. In contrast, F-18 FDG PET/CT is sensitive in staging evaluation of extracutaneous lesions regardless of CL or NCL.

Effects of Ampelopsis japonica Extracts on Tumor Immunity (백렴 추출물(抽出物)이 종양면역(腫瘍免疫)에 미치는 영향)

  • Park, Seung-Man;Ahn, Sang-Woo;Cho, Jong-Kwan
    • Korean Journal of Oriental Medicine
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    • v.11 no.2
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    • pp.113-140
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    • 2005
  • 1. 백렴 추출물은 부분적으로 종양세포인 HT1080, Hep G2, MCF-7 cell에 대해 세포독성을 보였으나, 섬유세포인 7250 cells과 종양세포인 CT-26 cells에서는 세포독성은 나타나지 않았다. 2. 백렴 추출물은 macrophage에서 생산되는 NO production을 부분적으로 증가시켰다. 3. 백렴 추출물은 macrophage 활성화와 관련 있는 IL-1과 iNOS의 유전자 발현을 증가시켰다. 4. 백렴 추출물은 NK cell의 cytotoxicity를 활성화시켰다. 5. 백렴 추출물은 NK cell 활성화와 관련있는 IL-1, IL-4, IL-10, IL-12, iNOS, IFN-${\gamma}$, TNF-${\alpha}$ 의 유전자 발현을 증가시켰다. 6. 백렴 추출물은 마우스의 비장세포에서 IL-10, IFN-${\gamma}$, TNF-${\alpha}$ 의 단백질 발현을 증가시켰다. 7. 백렴 추출물은 CT-26 cell에 의한 pulmonary colony를 대조군에 비해 유의성 있게 억제시켰다. 이상의 결과로 백렴 추출물은 macrophage와 NK cell의 활성화를 통해 면역 조절과 항암제로서의 유용성이 기대된다.

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Cytotoxic Activities of Green and Brown Seaweeds Collected from Jeju Island against Four Tumor Cell Lines

  • Kim, Kil-Nam;Lee, Ki-Wan;Song, Choon-Bok;Jeon, You-Jin
    • Preventive Nutrition and Food Science
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    • v.11 no.1
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    • pp.17-24
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    • 2006
  • Methanolic and aqueous extracts from 37 seaweed species (10 green and 27 brown seaweeds) collected from Jeju Island coast were prepared at high ($70^{\circ}C$) and room ($20^{\circ}C$) temperatures and examined for cytotoxic activity against 4 tumor cell lines: U937 (human monoblastoid leukemia cell line), HL60 (human promyelocytic leukemia cell line), HeLa (woman cervical carcinoma cell line) and CT26 (mouse colon carcinoma line). Both MeOH extracts of Desmarestia tabacoides and Dictyota dichotoma possessed strong cytotoxic activities against all the tumor cell lines tested, but the aqueous extract exhibited no activity. On the other hand Ecklonia cava showed strong cytotoxic activities for the $20^{\circ}C$ aqueous extract against the three tumor cells except HeLa cell. Sagassum coreanum and Sagassum siliquastrum $20^{\circ}C$ aqueous extracts also exhibited strong cytotoxic activities against U937, HL60, HeLa cells. Even though green seaweeds showed less activity than brown seaweeds, $20^{\circ}C$ aqueous extracts of Codium contractum and Codium fragile exhibited strong cytotoxic activities against HL60 or CT26 cells, respectively.

$^{99m}Tc$-HDP Bone Scintigraphy Finding of Metastatic Renal Cell Carcinoma Bone Lesion Changed from Hot to Cold Lesion: Comparing with $^{18}F$-FDG PET/CT ($^{99m}Tc$-HDP 뼈스캔의 열소에서 냉소로 변한 신세포암 뼈전이 소견: $^{18}F$-FDG PET/CT와의 비교)

  • Seo, Young-Duk;Kim, Seong-Min;Kim, Kun-Ho
    • Nuclear Medicine and Molecular Imaging
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    • v.43 no.6
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    • pp.588-591
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    • 2009
  • A 26-year-old man with renal cell carcinoma underwent $^{99m}Tc$-HDP bone scintigraphy for detecting bony metastasis after left total nephrectomy for renal cell carcinoma. $^{99m}Tc$-HDP bone scintigraphy showed small hot lesion in the first lumbar spine. About 12 months later, he underwent spinal MRI for lower back pain. A large mass was seen around spinous process of the first lumbar spine (L1) on spinal MRI and confirmed as metastatic renal cell carcinoma by bone biopsy. $^{99m}Tc$-HDP bone scintigraphy and $^{18}F$-FDG PET/CT were underwent for further evaluation. $^{99m}Tc$-HDP bone scintigraphy showed cold lesion in the first lumbar spine which was initially hot and newly developed hot lesion in the twelfth thoracic spine, and which were shown as hypermetabolic lesions in $^{18}F$-FDG PET/CT. We report a case of bony metastasis from renal cell carcinoma which is changed from hot lesion to cold lesion in $^{99m}Tc$-HDP bone scintigraphy and compare with $^{18}F$-FDG PET/CT.