(3.238.88.35) 您好!臺灣時間:2021/04/10 19:19
字體大小: 字級放大   字級縮小   預設字形  
回查詢結果

詳目顯示:::

我願授權國圖
: 
twitterline
研究生:劉昱含
研究生(外文):Liu, Yu-Han
論文名稱:利用磁振造影對比劑與螢光探針對肝癌原致癌因子ISX之分子影像研究
論文名稱(外文):Antibody conjugated MR-optical imaging agent (MnMEIO-(ISX)-CyTE777-mPEG nanoparticle) for in vivo and in vitro bio-imaging of oncoprotein intestine specific homeodomain on hepatocellular cancer
指導教授:王雲銘梁美智梁美智引用關係
指導教授(外文):Wang, Yun-MingLiang, Mei-Chih
口試委員:廖光文鄭添祿王雲銘梁美智
口試委員(外文):Liao, Kuang-WenCheng, Tian-LuWang, Yun-MingLiang, Mei-Chih
口試日期:2018-7-30
學位類別:碩士
校院名稱:國立交通大學
系所名稱:生物科技學系
學門:生命科學學門
學類:生物科技學類
論文種類:學術論文
論文出版年:2018
畢業學年度:106
語文別:英文
論文頁數:35
中文關鍵詞:肝細胞癌同源異型基因磁振造影超順磁氧化鐵奈米粒子
外文關鍵詞:Hepatocellular carcinomaIntestine specific homeoboxMagnetic resonance imagingSPIOIVIS
相關次數:
  • 被引用被引用:0
  • 點閱點閱:35
  • 評分評分:系統版面圖檔系統版面圖檔系統版面圖檔系統版面圖檔系統版面圖檔
  • 下載下載:0
  • 收藏至我的研究室書目清單書目收藏:0
肝細胞癌為人類常見的肝癌,同時是致死率前十大的癌症。目前對於肝癌的發生路徑研究還並未非常明確,但已知肝纖維化為肝細胞癌最嚴重的致癌因子,約80~90%肝細胞癌患者同時患有肝纖維化。近期研究發現了一種新型肝癌原致癌基因ISX,此種基因會被肝纖維化相關細胞因子IL-6激發,並對癌細胞生長,轉移與免疫抑制等功能有調控的效果。對ISX進一步的研究可能可以對肝癌的致癌機制獲得突破性的了解,或對早期肝癌診斷提供協助。本團隊藉由結合氧化鐵奈米粒子、螢光顯影劑CyTE777與對ISX專一性抗體,合成出具有標靶化顯影功能的多功能顯影奈米粒子。合成出來的粒子大小約為20奈米,其在細胞實驗測試中顯示出良好的生物相容性與物理穩定度,並在之後的細胞影像測試中展現出良好的標靶化螢光顯影與磁振造影解析度強化功能。在異體腫瘤移植的裸鼠上同樣表現出良好的活體核磁共振影像解析度增強的效果。
Hepatocellular carcinoma or HCC is the most common primary liver cancer. Although detailed oncogenesis is unknown, cirrhosis is considered to be the highest risk factor for the development of HCC. About 80 to 90% HCC cases was reported to occurred in the setting of cirrhosis. Recent research suggests that intestine specific homeobox (ISX) is a proto-oncogene for hepatocellular carcinoma which is triggered by cirrhosis related cytokines IL-6. Also, ISX is reported to have regulator effect on multiple cancer related cell functions including proliferation, transformation and immunosuppression. Further research on ISX could be important for advancing knowledge about HCC oncogenic pathway and possible early diagnostic of hepatocellular carcinoma. Nanoparticles (MnMEIO-(ISX)-CyTE777-mPEG) with fluorescence dye and iron oxide nanoparticals that specifically targeting ISX homeodomain were synthesized. Diameter of this particle is identified as approximately 20 nm which is suitable to enter the cell. And was studied for in vivo and in vitro imagine property. The result demonstrated that this nanoparticle is able to target and mark ISX homeodomain in vitro and in vivo, and have confirmed functional on both optical and MR imiging contrast enhancement.
VII

Contents
摘 要 ..................................................................................................... I
Abstract ................................................................................................... III
致 謝 .................................................................................................... V
Contents ................................................................................................. VII
Table of Figures ...................................................................................... IX
List of abbreviations .............................................................................. XI
1. Introduction ........................................................................................ 1
1.1. Hepatocellular carcinoma ....................................................... 1
1.2. Diagnosis method of hepatocellular carcinoma .................... 1
1.3. Intestine specific homeobox (ISX) .......................................... 2
1.4. MR imaging .............................................................................. 3
1.5. MnMEIO nanoparticle............................................................ 4
2. Project Aim ......................................................................................... 6
3. Material and Method ......................................................................... 7
3.1. Instrumentation ....................................................................... 7
3.2. Reagents .................................................................................... 7
3.3. Antibodies ................................................................................. 8
3.4. Animals ..................................................................................... 8
3.5. Methods .................................................................................... 8
3.5.1. Cell culture ............................................................................ 8
3.5.2. Whole cell lysates .................................................................. 9
3.5.3. Protein assay ......................................................................... 9
3.5.4. SDS-polyacrylamide gel electrophoresis ............................ 9
3.5.5. Western blot analysis ......................................................... 10
3.5.6. Confocal Microscopy.......................................................... 11
3.5.7. Confocal microscopy using living cell .............................. 11
3.5.8. Synthesis of MnMEIO-(ISX)-CyTE777-mPEG
nanoparticle ...................................................................................... 11
3.5.9. Dynamic light scattering analysis ..................................... 12
3.5.10. UV absorbance analysis ..................................................... 12
3.5.11. Cell viability ........................................................................ 12
3.5.12. Prussian blue stain ............................................................. 13
3.5.13. Cell preparation for in vitro optical and MR imaging ... 13
3.5.14. In vivo MR imaging study ................................................. 14

4. Results and Discussion ..................................................................... 14
4.1. The property of tetracycline inducible ISX system of
Sk-Hep1 cell ...................................................................................... 15
4.2. Characterization of MnMEIO-(ISX)-CyTE777-mPEG
nanoparticles..................................................................................... 18
4.3. MnMEIO-(ISX)-CyTE777-mPEG nanoparticle has no cell
toxicity ............................................................................................... 21
4.4. MnMEIO-(ISX)-CyTE777-mPEG nanoparticle is able to
specifically target ISX ...................................................................... 22
4.5. MnMEIO-(ISX)-CyTE777-mPEG nanoparticle can target
and mark ISX in cytoplasm of living cells ..................................... 24
4.6. In vivo MRI imaging analysis ............................................... 30
5. Conclusion......................................................................................... 32
6. References ......................................................................................... 33
33

6. References
[1] J. Balogh, D. Victor, E.H. Asham, S.G. Burroughs, M. Boktour, A. Saharia,
X. Li, R.M. Ghobrial, H.P. Monsour, Hepatocellular carcinoma: a review,
Journal of Hepatocellular Carcinoma 3 (2016) 41-53.
[2] M. Kudo, Early Hepatocellular Carcinoma: Definition and Diagnosis, Liver
Cancer 2(2) (2013) 69-72.
[3] D. Janevska, V. Chaloska-Ivanova, V. Janevski, Hepatocellular Carcinoma:
Risk Factors, Diagnosis and Treatment, Open Access Macedonian Journal of
Medical Sciences 3(4) (2015) 732-736.
[4] I. Choi Byung, The current status of imaging diagnosis of hepatocellular
carcinoma, Liver Transplantation 10(S2) (2004) S20-S25.
[5] J.Y. Choi, S.W. Jung, H.Y. Kim, M. Kim, Y. Kim, D.G. Kim, E.-J. Oh,
Diagnostic value of AFP-L3 and PIVKA-II in hepatocellular carcinoma
according to total-AFP, World Journal of Gastroenterology : WJG 19(3) (2013)
339-346.
[6] Y.X.J. Wáng, J.-M. Idée, A comprehensive literatures update of clinical
researches of superparamagnetic resonance iron oxide nanoparticles for
magnetic resonance imaging, Quantitative Imaging in Medicine and Surgery 7(1)
(2017) 88-122.
[7] E. Boncinelli, Homeobox genes and disease, Current Opinion in Genetics &
Development 7(3) (1997) 331-337.
[8] M. Levine, T. Hoey, Homeobox proteins as sequence-specific transcription
factors, Cell 55(4) (1988) 537-40.
[9] Y. Seino, T. Miki, H. Kiyonari, T. Abe, W. Fujimoto, K. Kimura, A. Takeuchi,
Y. Takahashi, Y. Oiso, T. Iwanaga, S. Seino, Isx participates in the maintenance
34

of vitamin A metabolism by regulation of beta-carotene 15,15'-monooxygenase
(Bcmo1) expression, J Biol Chem 283(8) (2008) 4905-11.
[10] L.T. Wang, S.S. Chiou, C.Y. Chai, E. Hsi, K.K. Yokoyama, S.N. Wang, S.K.
Huang, S.H. Hsu, Intestine-Specific Homeobox Gene ISX Integrates IL6
Signaling, Tryptophan Catabolism, and Immune Suppression, Cancer Res 77(15)
(2017) 4065-4077.
[11] S.N. Wang, L.T. Wang, D.P. Sun, C.Y. Chai, E. Hsi, H.T. Kuo, K.K.
Yokoyama, S.H. Hsu, Intestine-specific homeobox (ISX) upregulates E2F1
expression and related oncogenic activities in HCC, Oncotarget 7(24) (2016)
36924-36939.
[12] S. Sue, W. Shibata, E. Kameta, T. Sato, Y. Ishii, H. Kaneko, H. Miwa, T.
Sasaki, T. Tamura, M. Kondo, S. Maeda, Intestine-specific homeobox (ISX)
induces intestinal metaplasia and cell proliferation to contribute to gastric
carcinogenesis, J Gastroenterol 51(10) (2016) 949-60.
[13] A. Berger, Magnetic resonance imaging, BMJ : British Medical Journal
324(7328) (2002) 35-35.
[14] S. Tocchio, B. Kline-Fath, E. Kanal, V.J. Schmithorst, A. Panigrahy, MRI
Evaluation and Safety in the Developing Brain, Seminars in perinatology 39(2)
(2015) 73-104.
[15] M.J. Knight, S. Dillon, L. Jarutyte, R.A. Kauppinen, Magnetic Resonance
Relaxation Anisotropy: Physical Principles and Uses in Microstructure Imaging,
Biophysical Journal 112(7) (2017) 1517-1528.
[16] L. Valkovič, M. Chmelík, M. Krššák, In-vivo(31)P-MRS of skeletal muscle
and liver: A way for non-invasive assessment of their metabolism, Analytical
Biochemistry 529 (2017) 193-215.
[17] D.J. Pennell, A.J. Baksi, S.K. Prasad, R.H. Mohiaddin, F. Alpendurada, S.V.
35

Babu-Narayan, J.E. Schneider, D.N. Firmin, Review of Journal of
Cardiovascular Magnetic Resonance 2015, Journal of Cardiovascular Magnetic
Resonance 18 (2016) 86.
[18] X.-M. Zhu, Y.-X.J. Wang, K.C.-F. Leung, S.-F. Lee, F. Zhao, D.-W. Wang,
J.M.Y. Lai, C. Wan, C.H.K. Cheng, A.T. Ahuja, Enhanced cellular uptake of
aminosilane-coated superparamagnetic iron oxide nanoparticles in mammalian
cell lines, International Journal of Nanomedicine 7 (2012) 953-964.
[19] P.-C. Liang, Y.-C. Chen, C.-F. Chiang, L.-R. Mo, S.-Y. Wei, W.-Y. Hsieh,
W.-L. Lin, Doxorubicin-modified magnetic nanoparticles as a drug delivery
system for magnetic resonance imaging-monitoring magnet-enhancing tumor
chemotherapy, International Journal of Nanomedicine 11 (2016) 2021-2037.
[20] W. Zhang, L. Liu, H. Chen, K. Hu, I. Delahunty, S. Gao, J. Xie, Surface
impact on nanoparticle-based magnetic resonance imaging contrast agents,
Theranostics 8(9) (2018) 2521-2548.
[21] C. Gao, Z.-J. Deng, D. Peng, Y.-S. Jin, Y. Ma, Y.-Y. Li, Y.-K. Zhu, J.-Z. Xi,
J. Tian, Z.-F. Dai, C.-H. Li, X.-L. Liang, Near-infrared dye-loaded magnetic
nanoparticles as photoacoustic contrast agent for enhanced tumor imaging,
Cancer Biology & Medicine 13(3) (2016) 349-359.
[22] B. Heim, F. Krismer, R. De Marzi, K. Seppi, Magnetic resonance imaging
for the diagnosis of Parkinson’s disease, Journal of Neural Transmission 124(8)
(2017) 915-964.
電子全文 電子全文(網際網路公開日期:20211002)
連結至畢業學校之論文網頁點我開啟連結
註: 此連結為研究生畢業學校所提供,不一定有電子全文可供下載,若連結有誤,請點選上方之〝勘誤回報〞功能,我們會盡快修正,謝謝!
QRCODE
 
 
 
 
 
                                                                                                                                                                                                                                                                                                                                                                                                               
第一頁 上一頁 下一頁 最後一頁 top
系統版面圖檔 系統版面圖檔