硝酸鎵調節SK Hep-1細胞鈣磷脂膜聯蛋白A2分子的表現

Gallium Nitrate Regulates Annexin A2 Expression in SK Hep-1 Cells

蔡孟達(Meng-Ta Tsai)

指導教授 ： 林詠峯

臺北醫學大學/醫學科技學院/醫學檢驗暨生物技術學系所/碩士(2014年)

https://doi.org/10.6831/TMU.2014.00050

摘要

鎵是兩性金屬元素，具有自然界所有元素中沸點與熔點差異值最大的特殊物理性質，因此，使得它應用廣泛。根據研究文獻記載，在細胞生理學上，鎵類似鐵，經轉鐵蛋白運送與鐵競爭結合位置。在其他研究也指出，鎵可經某些機制去影響鈣離子在細胞內的活性。鎵是否還會透過其他機轉與細胞其他的蛋白作用，或影響細胞其他生理功能，目前的研究甚少。本實驗以具解毒功能的肝細胞株 (SK Hep-1)作為體外研究模型，以硝酸鎵 [Ga(NO3)3] 來做為取得鎵離子的來源；然後我們使用液相層析-串聯式質譜儀 (LC MS/MS) 作為蛋白篩選分析工具。實驗中篩選了該細胞株中近九百種蛋白，通過質譜儀將胜肽分析比對之後，再以西方墨點法 (Western blotting)與反轉錄-聚合酶連鎖反應 (RT-PCR)作驗證，來尋找出細胞緩解鎵造成生長壓力的機制。在此次研究發現，鎵減低了鈣磷脂膜聯蛋白A2 (annexin A2)的表達，這可能影響了細胞內鈣的調節功能。總而言之，在細胞中鎵可能透過鈣相關蛋白緩解了鎵離子對細胞生長的壓力，也調節了細胞內鈣離子的代謝。
關鍵字:鈣;鎵;鈣磷脂膜聯蛋白A2;高血鈣症;硝酸鎵

關鍵字

鈣 ； 鎵 ； 鈣磷脂膜聯蛋白A2 ； 高血鈣症 ； 硝酸鎵

並列摘要

Abstract
Gallium is an amphoteric metal. It has special physical properties of high boiling point and low melting point, making it widely used. Based on current research literature in cell physiology, gallium is similar to iron, and can be transported via transferrin, competing with the iron binding sites. Other reports indicate that gallium affects intracellular calcium activities through some mechanisms. It is not clear that whether gallium is also transferred through other mechanisms with other pro-teins, or it affects other physiological function of the cells. In this study, we used the detoxifying liver cell line SK Hep-1 as an in vitro model and Ga(NO3)3 as the gallium source. Ga(NO3)3 is a U.S. FDA-approved drug for the treatment of hypercalcemia of malignancy diseases. Liquid chromatography-tandem mass spectrometry (LC MS/MS) was used as a protein screening tool. In the experiment, we screened nearly nine hun-dred kinds of protein in the cells. After the spectrometric analysis of peptides, we used western blotting and reverse transcriptase-polymerase chain reaction (RT-PCR) to verify the data and find out the mechanisms of detoxifying gallium on growth pressure. We discovered that gallium reduced annexin A2 expression, which may affect calcium-associated proteins, and may regulate calcium metabolism in the cells.
Keywords:calcium;gallium;annexin A2;hypercalcemia;Ga(NO3)3

並列關鍵字

calcium ； gallium ； annexin A2 ； hypercalcemia ； Ga(NO3)3

參考文獻

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