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研究生：	陳冠維 Chen, Kuan-Wei
論文名稱：	應用舒曼波於人類乳腺癌細胞抑制效果之研究 Study of Inhibition of Human Breast Cancer Cells Using Schumann Wave
指導教授：	張凌昇 Jang, Ling-Sheng
學位類別：	碩士 Master
系所名稱：	電機資訊學院 - 電機工程學系 Department of Electrical Engineering
論文出版年：	2018
畢業學年度：	106
語文別：	英文
論文頁數：	56
中文關鍵詞：	電磁場、參數調變、細胞存活率、乳腺癌細胞
外文關鍵詞：	EMF, Parameters modulation, Cell viability, MDA-MB-231
相關次數：	點閱：57 下載：0
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腫瘤是世界上致死率最高的疾病之一，而其最主要的治療方式包括手術、化療、放射線治療與標靶治療，然而僅有少數的病患能在接受上述治療後成功從癌症中痊癒，此外這些治療方式均具有如噁心、暈眩或免疫力下降等嚴重的副作用，使許多患者長期飽受其苦。
許多研究團隊在實驗中發現，暴露於特定參數的電磁場使得腫瘤細胞活性降低，甚至抑制腫瘤細胞的增生。現今相關理論大多建立在電磁場與細胞內鈣離子產生共振現象，進而導致細胞加速凋亡，惟究竟電磁場抑制癌細胞生長的關鍵參數為何，時至今日尚未有切確的解釋。基於以上理由，本研究嘗試利用地球最原始的頻率－7.83 Hz舒曼共振頻率結合調變不同的電磁場參數來探討其對人類乳腺癌細胞的影響性。
實驗結果顯示，MDA-MB-231乳癌細胞暴露在7.83 Hz且掃頻區間為0.3Hz的脈衝電磁場連續48小時，以及切換暴露6小時並持續兩天之後，有顯著的細胞活力抑制現象。此外，加大線圈磁場後，乳腺癌細胞活力更是隨著加大的磁場而有大幅度地減少，但對正常乳腺細胞卻沒有影響，故作者推論癌細胞對於電磁場的抑制反應是參數依賴性的、細胞特異性的、多維度參數共同影響的。

Cancer is one of the most common causes of death in this world. Existing treatments such as radiotherapy and chemotherapy are associated with many side effects and only a low percentage of patients can be fully recovered. Many research teams have found that exposure to a specific-modulated EMF decreases the cell viability and even inhibits the proliferation of tumor cells.
Current theories are based on the resonance between EMF and intracellular calcium ions, which accelerates cell apoptosis and inhibits the differentiation or proliferation of cells. While evidence about anti-cancer effects exists, the critical parameter causing this effect is still unknown. Base on above reasons, we attempt to use Schumann resonance frequency of 7.83 Hz, combining with other modulated parameters to expose breast cancer MDA-MB-231 cells.
Breast cancer cells were exposed to the EMF with a frequency of 7.83 Hz and a sweep interval of 0.3 Hz for different temporal parameters. Also, the current flowing through coil windings was modulated up to 60 mA to investigate the effectiveness of EMF therapy on cancer cells. The results show that there was a significant reduction of cell number. Then we inference that the inhibitory effect is dependent on vatious parameters, and is multi-dimensional and cell-specific.

Contents
中文摘要……………………………………………………….……………………I
Abstract………………………………………………………………………………II
Acknowledgement…………………………………………………………………III
Contents……………………………………………………………………………IV
List of Table………………………………………………………………………..VII
List of Figure………………………………………………………………………VIII
Chapter 1 Introduction………………………………………………….1
    1-1 Background and Motivation…….………………………………………...…1
1-1-1 Background……………………………………………………………...1
1-1-2 Motivation………………………………………………………………2
    1-2 Review of related works…………………………………………...…………3
    1-3 Dissertation organization………………………………………...…………6

Chapter 2 Physical mechanisms of EMF exposure on cells……………7
    2-1 Biological system …………………………………………………………..7
      2-1-1 Cell system…………………………………………………………….7
      2-1-2 Electrical equivalent circuit for cell…………………………………….8
    2-2 Electrophysiological Theory………………………………………………10
      2-2-1 Ionic current………………………………………………………….10
      2-2-2 Membrane potential depolarization…………………………………….12
      2-2-3 Voltage-gated calcium channel……………………………………….14
    2-3 Molecular biophysics theories……………………………………………..16
      2-3-1 Orientation of electic dipoles……………………………………..……16
      2-3-2 Orientation of magnetic dipoles………………………………………..17
      2-3-3 Energy-level transition and splitting…………………………………...19    
      2-4 Cyclotron resonance frequency…………………………………………..21

Chapter 3 Materials and Methods…………………………………..23
    3-1 Cell cultures…………………………………………………………………23
      3-1-1 Cell line………………………………………………………………23
3-1-2 Cell culture…………………………………………………………...23
    3-2 Device of EMF exposure system……………………………………………24
      3-2-1 Square wave generator……………………………………………….24
      3-2-2 Coil windings………………………………………………………….24
      3-2-3 Noise shielding case………………………………………………….26
    3-3 Experimental setups and Parameters modulation…………………………..28
      3-3-1 Experimental setups……………………………………………..……28
      3-3-2 Signal type……………………………………………………………..30
      3-3-3 Applied frequency……………………………………………………...30
      3-3-4 Magnitude of magnetic field…………………………………………...31
      3-3-5 Exposure time………………………………………………………...32
    3-4 Measurement and Analysis Methods……………………………………..33
      3-4-1 MTT assay………………………………………………………..……33
      3-4-2 Optical density value…………………………………………………..33
      3-4-3 Statistical analysis……………………………………………………...34

Chapter 4 Results and Discussion……………………………………35
    4-1 Cell viability……………………………………………………………….35
    4-2 Results…………………………………………………………………….35
      4-2-1 Results of continuous exposure on MDA-MB-231……………………35
      4-2-2 Results of switching exposure on MDA-MB-231……………………39
      4-2-3 Results of EMF-magnitude modulation on breast cells………………..41
    4-3 Discussions…………………………………………………………………43

Chapter 5 Conclusions and Future Works…………………….…...49

References………………………………………………………….…...50


List of Table
Table 1-2-1. In vitro studies of EMF exposure in oncology…………………………5
Table 1-2-2. In vitro studies of EMF exposure on human breast cancer cells………5
Table 4-3-2. Expression of VGCCs in various human breast cell lines.…………...47


List of Figure
Fig. 2-1-1. Basic structure of a single cell……………………………………………7
Fig. 2-1-2. (a) Basic structure of cell plasma membrane (b) Electrical equivalent circuit 
         for cell membrane…………………………………………………………9
Fig. 2-1-3. Electrical equivalent circuit with built-in potential for cell membrane……10
Fig. 2-2-1. The formation of ion current under an EMF………………………………11
Fig. 2-2-2. Dynamic equilibrium of charged particles across the cell membrane……13
Fig. 2-2-3. Protein structure of Voltage gated ion channels…………………………14
Fig. 2-2-4. The analogy between the VICC and a FET………………………………15
Fig. 2-2-5. Difference in activated voltage between L-type and T-type VGCCs……16
Fig. 2-3-1. Atomic energy level distribution…………………………………………20
Fig. 3-2-1. Block diagram of in vitro exposure system……………………..……...24
Fig. 3-2-2. The geometry of multi-turns coil winding………………………………25
Fig. 3-2-3. (a) Top view of magnetic field distribution within the coil (b) Side view of  
          magnetic field distribution………………………………………………25
Fig. 3-2-4. The relationship between reflection loss, absorption loss, and multiple  
          reflection loss……………………………………………………………26
Fig. 3-2-5. The shielding device using in this study………………………………… 27
Fig. 3-3-1. Roadmap of experimental grouping method……………………………28
Fig. 3-3-2. Exposure experiment process……………………………………………29
Fig. 3-3-3. FFT analysis of sweep frequency 7.83±0.3 Hz for 0.1 Hz beating steps…30
Fig. 3-3-4. Based on the field magnitude and field uniformity, 96-well culture plate are 
         divided into 5 different position groups…………………………………32
Fig. 3-4-1. Liquid in quartz tubing……………………………………………………34
Fig. 4-2-1. Cell viability of invasive human breast cancer cell line MDA-MB-231 under the continuous exposure to EMF with a frequency of 7.83±0.3 Hz and coil current of 32 mA for 12 hours…………………………………………35
Fig. 4-2-2. Cell viability of human breast cancer cell line MDA-MB-231 under the 
          continuous exposure of EMF for 24 hours……………………………37
Fig. 4-2-3. Cell viability of human breast cancer cell line MDA-MB-231 under the  
          continuous exposure of EMF for 48 hours……………………………38
Fig. 4-2-4. Cell viability of human breast cancer cell line MDA-MB-231 under the 
          switching exposure of EMF for 6 hours………………………………39
Fig. 4-2-5. Cell viability of human breast cancer cell line MDA-MB-231 under the continuous exposure of EMF for 12 hours……………………………40
Fig. 4-2-6. Cell viability of human breast cancer cell line MDA-MB-231 under the 
          switching exposure of the EMF with 60mA coil current for 6 hours…41
Fig. 4-2-7. Cell viability of human breast cancer cell line M10 under the switching exposure of the EMF with 60mA coil current for 6 hours……………42
Fig. 4-3-1. Cell viability of MDA-MB-231 cells under various parameters modulation 
          at position 2…………………………………………………………43
Fig. 4-3-2. Cell viability of MDA-MB-231 cells under various parameters modulation 
          at position 5…………………………………………………………44

                                    

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