背景：口腔鱗狀細胞癌為我國非常盛行之癌症，並以男性居多。若能及早診斷，了解其病變過程及基因變化，對於控制此一疾病將極有助益。口腔鱗狀細胞癌是一種預後很差的癌症，在過去二十年口腔癌的平均五年存活率並沒有顯著的改善，在腫瘤形成中，血管新生是一個相當重要的機轉，其中會影響的因子，包含血小板衍生性生長因子(Platelet Derived Growth Factor(PDGF))。根據先前研究發現PDGF基因，在頭頸癌及其他癌症的癌細胞組織中，有過度表現及術後存活率較低，而PDGF在口腔鱗狀細胞癌之生成及進行角色未明。PDGF也和細胞週期調控及細胞凋零有關，在目前的研究中顯示，PDGF及其受體基因表現和許多疾病、癌症的進程有關。目的：本實驗為探討PDGF及其受體在口腔鱗狀細胞癌之表現，主要分析以血清含量中的PDGF、血小板增多症(Thrombocytosis)，及臨床參數之間的相關性，另外分析口腔鱗狀細胞癌組織及癌旁組織的表現量，是否有差異。最後再探討PDGF在，口腔鱗狀細胞癌細胞株中的功能，以做為診斷口腔癌的生物指標之可行性。研究方法：收集156位口腔鱗狀細胞癌病患，抽取手術前5毫升的血液樣本，利用酵素結合免疫吸附法(ELISA)分析PDGF-AA、BB血清濃度表現量，同時也分析血小板增多症與PDGF是否有關聯。再以統計方法分析，PDGF和臨床病理參數是否有相關性。另一方面利用免疫組織染色方法來探究，口腔鱗狀細胞癌組織之PDGF、PDGFR-α、β的表現量。並進一步利用定量反轉錄PCR(QRT-PCR)，分析112個口腔鱗狀細胞癌病人的，癌組織及癌旁組織的PDGF-AA、BB、PDGFR-α、β之基因表現量。以統計方法分析癌組織和癌旁組織，之基因表現是否有差異。最後在體外實驗，製造出過度表現PDGF之癌細胞株，探討PDGF過度表現，是否影響細胞功能。結果：我們的研究顯示術前PDGF-AA、BB與血小板有增多症有關聯(P<0.0001)，另外研究也顯示術前PDGF-BB與腫瘤大小(P=0.046)、頸部淋巴結轉移(P=0.016)、淋巴血管侵犯有相關性(P=0.005)。PDGF-BB (P=0.035)、血小板有增多症(P=0.018)與病患的存活率也是呈現正相關。另外研究也顯示PDGF-AA、BB及PDGFR-β之基因，在口腔鱗狀細胞癌組織，有顯著的過度表現(P<0.001)。在體外實驗我們發現PDGF可以促進 SAS與OECM-1的移行能力(Cell migration)、細胞群落生成(Colony formation) 以及細胞侵襲能力(Cell invasion )。結論：PDGF-AA、PDGF-BB及PDGFR-β基因在口腔鱗狀細胞癌組織，有顯著的過度表現，與臨床參數之間也有關聯性。在體外實驗也可以增加，癌細胞株生長、移行及侵襲能力。這樣的結果顯示出PDGF-AA、BB，在口腔鱗狀細胞癌，發展生成上應扮演重要的角色，另外我們發現過度表現的PDGF，使血中血小板數目增加。推測PDGF可能是藉由血小板的表現量增加，而快速新血管形成，進而增加細胞的移行能力，使癌細胞能夠轉移到身體其他部位，導致術後存活力降低。在口腔鱗狀細胞癌病患血清中，PDGF的表現量與一般健康人比較，PDGF的表現量較高，因此血液之PDGF具有作為口腔癌診斷指標之潛力，PDGFR-β在癌症組織的過量表現是較為新穎的發現，PDGFR-β所引發的相關機轉值得進一步探討。

Oral cancer is a kind of cancer with poor prognosis. In Taiwan, oral cancer is the 6th highest among all cancers in causes of death, and the mortality rate is the 4th highest in male. According to the statistics, the 5-year survival rate haven’t been improved significantly during past 20 years. Angiogenesis is one of the most important mechanism in tumorogenesis. In our studies, we conduct the study to clarify the relationship between the expression serum level of PDGF-AA and PDGF-BB with clinical and pathological parameters in OSCC patient. There were 156 OSCC patients were enrolled in this study. 5 ml blood was collected before operation for each patient. We estimated the pre-operative serum level of PDGF-AA and BB By using ELISA. In addition ,we compared PDGF-AA、BB、PDGFR-α、β mRNA expression between 112 pairs of oral squamous cell carcinoma (OSCC) tissue and non-cancerous matched tissue (NCMT). Finally, we discuss the function of PDGF in oral squamous cell carcinoma cell line to be used as a biomarker to diagnose oral cancer. The results revealed elevated PDGF-BB level has positive correlation with larger tumor size, neck lymph node metastasis, advanced TNM staging and positive perineural invasion. Thrombocytosis has positive correlation with larger tumor size, neck lymph node metastasis, advanced TNM staging, Furthermore, our study revealed elevated pre-operative serum PDGF-AA、BB、Thrombocytosis respectively has highly positive correlation with worse overall clinical outcome. PDGF-BB may has potential as useful tumor marker for predicting clinicopathological parameters and poor prognosis of OSCC, another result revealed significant over-expression of PDGF-AA 、 PDGF-BB and PDGFR-β mRNA in cancerous tissue, which implied that PDGF-AA and PDGF-BB might be the promising biomarkers for OSCC. In vitro study, we found that over-expressed PDGF can promoted cell proliferation, cell migration, cell invasion and colony formation for OECM-1 and SAS of cell line. In conclusion PDGF-AA, PDGF-BB and its receptor (PDGFR-β) have significant over-expression in oral squamous cell carcinoma, and were also related to clinical-pathologic parameters. In vitro study can also promoted increase the tumor cell growth ability of cancer cells line. This result shows that PDGF-AA, PDGF-BB, in the development of oral squamous cell carcinoma should play an important role. In addition, we found that over-expression of PDGF increased the number of platelets. Suggesting that PDGF may be due to the increase in the performance of platelets and rapid angiogenesis, and thus increase the ability of cell migration, so that distant metastasis of cancer cells to other parts of the organ, resulting in poor survival rate. The expression of PDGF in serum of patients with oral cancer is higher than that of healthindividuals.Therefore, the PDGF of blood has the potential as a diagnostic indicator of oral cancer, and the over-expression of PDGFR-β in cancer tissue is relatively novel Found. Further research would be necessary to clarify the role of PDGFR expression with tumor cell behavior in OSCC.

中文摘要…………………………………………………… ………… i
Abstract……………………………………………………………….iv
目錄…………………………………………………………………….vii
圖次目錄……………………………………………………………….viii
表次目錄………………………………………………………………..ix
緒論……………………………………………………………………..1
研究目標………………………………………………………………..8
研究材料與方法………………………………………………………..9
統計…………………………………………………………………….21
結果…………………………………………………………………….22
討論…………………………………………………………………….26
結論…………………………………………………………………….34
圖列…………………………………………………………………….35
表列…………………………………………………………………….50
參考文獻……………………………………………………………….59
附錄…………………………………………………………………….71

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