由三種不同危險因子衍生之口腔癌細胞株定量磷酸化蛋白體差異之探討

Quantitative phosphoproteome of oral cancer cell lines associated with three different risk factors

張華元(Hua-Yuan Chang)

指導教授 ： 陳逸夫

高雄醫學大學/生命科學院/生物科技學系碩士班/碩士(2012年)

摘要

蛋白質酪胺酸磷酸化調節許多細胞功能，也已經被證實是參與癌變重要的一門關鍵。因此細胞內蛋白質的酪胺酸磷酸化（pTyr）的鑑定是一個非常重要的任務。由於酪氨酸磷酸化的表現佔總體磷酸化只約1％，酪胺酸磷酸化蛋白質的研究是相當具有挑戰性。口腔鱗狀細胞癌（OSCC），是最普遍的口腔惡性腫瘤，已成為台灣男性常見的前五大癌症之一。常見的口腔癌致癌風險因子為(A)飲酒、(B)嚼食檳榔和(C)吸煙，這些都是在台灣口腔癌發病的主要危險因素。然而，這三種風險因子誘發口腔癌之機轉尚未闡明。由於致癌信號途徑中蛋白磷酸化扮演重要的關鍵，在這項研究中，我們試圖找出不同風險因子衍生之口腔癌相關的磷酸化蛋白; ABC-free，B-only和ABC。目前我們已經成功的建立了該三株不同風險因子所誘發的口腔癌細胞株。此外，建立單獨吸煙病史(C-only)的口腔鱗狀癌細胞株亦正在進行中。我們利用iTRAQ標記磷酸化蛋白質並以質譜做為基礎的標記定量蛋白質體學實驗平台，未來我們將釐清不同風險因子所衍生之口腔癌間磷酸化蛋白差異與功能性研究。

關鍵字

磷酸化 ； 口腔鱗狀細胞癌 ； iTRAQ

並列摘要

Phosphorylation of protein tyrosine residues regulates many cell functions and has also been proved to be involved in oncogenesis. Thus, identification of the phosphotyrosine (pTyr) proteome of cells is a very important task. Since tyrosine phosphorylation represents only around 1% of the total human phosphoproteome, the study of pTyr proteins is rather challenging. Oral squamous cell carcinoma (OSCC), the most widespread malignant neoplasm of the oral cavity, has become the fifth common cancer in Taiwanese male population. It is generally believed that oral habits, including alcohol drinking (A), betel quid chewing (B) and cigarette smoking (C), are major risk factors of oral cancer incidence. However, how these three risk factors induce oral cancer has yet to be elucidated. Since protein phosphorylation plays a critical role in oncogenic signaling pathways, in this study, we sought to identify the phosphoproteome of three oral cancer cell lines associated with the risk factors; ABC-free, B only and ABC. Currently we have established three oral cancer cell lines associated with three different risk factors. Besides, establishment of a smoking-only (C only) OSCC cell line is ongoing. We have also built up a phosphotyrosine peptides enrichment workflow coupled with mass spectrometry-based iTRAQ or label-free quantitative proteomics. In the future, we will finish quantitative phosphotyrosine proteome of oral cancer cell lines with such three combinations of risk factors and functional studies of the candidate phosphotyrosine proteins.

並列關鍵字

Phosphorylation ； OSCC ； iTRA

參考文獻

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