果糖二磷酸醛縮酶C於口腔鱗狀細胞癌進程之效應

The Effect of Fructose-bisphosphate Aldolase C in Oral Squamous Cell Carcinoma Progression

黃紫虹(Tse-Hung Huang)

指導教授 ： 張正琪

國立臺灣大學/牙醫專業學院/口腔生物科學研究所/碩士(2014年)

https://doi.org/10.6342/NTU.2014.10623

摘要

目的:
糖解作用中的醣解酶表現異常會調控腫瘤細胞的行為。然而，醣解酶調控口腔鱗狀細胞癌進程之角色仍尚未釐清。
材料方法:
利用即時聚合酶鏈鎖反應檢測果醣二磷酸醛縮酶C之mRNA表現量與口腔癌病人檢體之關係。進一步利用抑制或過度表現果醣二磷酸醛縮酶C基因在口腔鱗狀細胞癌細胞株的方法證實細胞爬行或轉移的能力，並建立原位注射口腔癌的動物模型來研究果醣二磷酸醛縮酶C在動物模式內淋巴結轉移與存活率的結果。
結果:
我們首先確定了在惡性程度較高的細胞株，SAS及Ca9-22細胞中果醣二磷酸醛縮酶C的表現量顯著地較低。臨床資料顯示，在68個口腔癌病人中，果糖二磷酸醛縮酶C的表現量與淋巴結轉移和早期的病人檢體中呈負相關性。過度表現果醣二磷酸醛縮酶C顯著地抑制ATP和乳酸的生成，導致細胞爬行能力降低；抑制果醣二磷酸醛縮酶C可以回復細胞侵襲及爬行的能力。此外，催化糖解作用之位點Arg42和K146進行單點突變及雙突變，並將突變的果醣二磷酸醛縮酶C轉染至SAS內，可以回復其抑制細胞侵襲及爬行之能力。在動物模型中證實果醣二磷酸醛縮酶C過度表現顯著地降低淋巴結的轉移並延長動物的存活率。
結論:
果醣二磷酸醛縮酶C為一個口腔鱗狀細胞癌臨床診斷及預後標記的重要角色，其功能為抑制口腔鱗狀細胞癌侵襲及爬行之能力，而Arg42及Lys146為重要調控的位點。本論文顯示，果醣二磷酸醛縮酶C可能是潛在治療標靶以阻止口腔鱗狀細胞癌的進程。

關鍵字

口腔癌 ； 轉移 ； 果糖二磷酸醛縮酶C ； 糖解酶 ； 代謝

並列摘要

Objectives:
Glycolysis machinery regulates cancer cell behavior. However, the role of these glycolysis enzyme in oral squamous cell carcinoma (OSCC) progression is still largely unknown.
Materials and Methods:
Fructose-bisphosphate aldolase C (ALDOC) expression in OSCC patients and cell lines was detected by real-time quantitative RT-PCR. The functions of ALDOC in migration and invasion ability of OSCC cells were determined by gain and loss function approaches. The orthotopic OSCC animal model was performed to investigate the effects of ALDOC on OSCC cell lymph node metastasis and tumorigensis in vivo.
Results:
ALDOC mRNA and protein expression was significantly decreased in advanced migratory cells including SAS and Ca9-22 cells. Clinical data revealed that ALDOC mRNA expression negatively correlated with lymph node metastasis and advanced tumor TNM stage in 68 OSCC patients. Overexpressed ALDOC block ATP generation and lactate production resulted in cell motility retardation. Transfected silenced ALDOC expression plasmid could restore OSCC cell migration and invasion.
Furthermore, Transfection of ALDOC point mutation constructs of catalytic domain, Arg42 and K146 in SAS cells could functionally restore the ALDOC-inhibited cell migration and invasion. In vivo, ALDOC overexpressed significantly decreased lymph node metastasis and prolong mice survival.
Conclusion:
ALDOC plays as an OSCC prognosis marker clinically, and functions to suppressor OSCC cell migration and invasion, and the catalytic domain of Arg42 and K146 is crucial in this regulations machinery. ALDOC could be a potential therapeutic target to block OSCC progression.

並列關鍵字

oral cancer ； migration ； ALDOC ； glycolysis enzyme ； metabolism

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