| Abstract: | 由行政院衛生福利部最新資料顯示，口腔癌是臺灣地區發生率和死亡率增加速度最快之癌症。因此尋找可快速精準診斷口腔癌的生物標記 (biomarker) 及了解口腔癌形成之分子機轉，以期能早期診斷口腔癌並給予適當的治療，減少口腔癌之發生和增加口腔癌之存活率，是當前重要的研究課題。本分項計畫之子計畫一發現部分微量菌種在不同癌症相關因子 (菸、酒、檳榔等) 與檢體分組分析得到顯著差異，這些微量菌可以單一的對不同分組的檢體做出區隔，但與其他菌種及其他癌症相關因子間的關係並不明確。因此我們引入新的演算法來分析，希望可以找到相互加強分辨率的數種因子，以作為判定癌症病變的可能組合，並且達到 90% 以上的預測準確率，並以其中的菌株組合作為判定口腔癌的生物標誌。本子計畫二已經建立一個包含9個基質金屬蛋白酶(Matrix metalloproteinases; MMPs) (MMP-1、MMP-2、MMP-3、MMP-7、MMP-8、MMP-9、MMP-10、MMP-12、MMP-13)及4個TIMPs抑制劑(TIMP-1、TIMP-2、TIMP-3及TIMP-4)的蛋白質晶片。並利用59個正常人的唾液檢體及50個口腔癌的唾液檢體，再搭配其性別、年齡、抽菸、喝酒及嚼食檳榔的習慣。利用生物統計分析發現，當同時具有7個變數時，可以達到最好的正確率。我們也發現血漿中高表現的MMP-11濃度與口腔癌患者的期數、腫瘤大小及近端淋巴轉移有明顯的相關性，其結果也發表PLos ONE 期刊。我們也進一步分析TIMP-3 rs9862此位置的基因多型性與血漿中TIMP-3濃度的相關性，結果發現，高表現的TIMP-3與口腔癌患者的腫瘤大小有明顯的相關性，並與其基因型的分布有關。本子計畫三發現相較於正常口腔上皮組織，微核糖核酸miR-204於口腔癌腫瘤組織及其淋巴轉移組織低度表現; 且第三、四期口腔癌癌化組織之miR-204的表現量低於第一、二期口腔癌癌化組織。miR-204於口癌癌幹細胞(幹細胞球體、子族群細胞、及ALDH1正表現細胞)低度表現;因此，我們發現過度表現miR-204可降低癌幹細胞球體自我更新能力、細胞轉移/侵襲力、細胞形成群落之能力。利用裸鼠異種移植模式發現過度表現miR-204可抑制口腔癌幹細胞活體腫瘤生長的體積。利用螢光酵素分析及西方墨點法驗證Slug及Sox4為miR-204之直接標靶基因。miR-204表現抑制亦可增加口腔癌細胞形成幹細胞球體能力及細胞轉侵襲力。利用裸鼠原位異種移植模型發現miR-204表現抑制可促使口腔癌細胞淋巴結轉移。結合細胞、動物模型、及臨床組織推測腫瘤抑制型微核糖核酸miR-204可調控癌幹細胞特性而影響口腔癌致病機轉，期待應用於口腔癌淋巴轉移的診治及監測。本子計畫四發現在接受過放射性治療的2 例口腔癌病人檢體中，佔據最多表現量的特定少數 TRBV 與 TRBJ 其數量雖有變化，但組合並沒有改變。故而推論因口腔癌組織存在免疫系統的相關反應可能還未及恢復或消除。
Oral cancer (OC) has been the 4th leading cause of cancer death among males in Taiwan. To date, more than 60% of OC patients are diagnosed as locally advanced cancer stage, which account for poor prognosis. There is an urgent need to develop an effective screening protocol and treatment policy to lower the incidence of OC and to promote these patients’ survival. In the Theme 1: We found some minor bacteria showing significantly different distribution between each group (Normal, Precancer, and Cancer) or some factors (smoking, alcohol, and betel). These strands can distinguish samples from different groups. However, the relationships between these bacteria and factors are still unknown. We are introducing new algorithm to analyze groups of factors and trying to find the best groups that can have highest specificity and sensitivity. Those bacteria included in the group could be the novel biomarker of oral cancer. In the Theme 2: We developed a protein chip following the biomarkers validated (including MMP-1, MMP-2, MMP-3, MMP-7, MMP-8, MMP-9, MMP-10, MMP-12, MMP-13) for multi-analyte approach. We found the AUC of seven features (MMP-3、MMP-10、MMP-13、TIMP-2, age, gender and betel quid chewing) was 0.916. We also showed that a substantial increase in the plasma level of MMP-11 is useful for assessment of the disease progression, especially lymph node metastasis, in patients with oral cancer. These results have been published in PLoS ONE. Moreover, TIMP3 plasma levels significantly increased in oral cancer patients who have large tumor or carry T allele rs9862 polymorphism. In the Theme 3: Compared with non-tumor samples from the same patient, the expression of miR-204 was decreased in all of the tumor samples. A similar down-regulation of miR-204 was also observed in metastatic lymph nodes when compared with local tumors. Lower miR-204 expression was detected in oral cancer stem cells (CSCs). Overexpression of miR-204 dramatically alleviated CSCs properties and tumor progression in CSCs-transplanted mice. Reporter assays further revealed that miR-204 directly targets the 3' UTR regions of Slug and Sox4. MiR-204 may be a useful diagnostic oral cancer tumor marker and novel miRNA-based approach for oral cancer treatment. In the Theme 4: We found the major TRBV and TRBJ domains in the two samples from radiotherapy treated patients are still the same as from before treatment. Also the D50 value did not show the trend to recover. Therefore, we thought we may need to increase the number of check points after treatment to see the recovery of immune system. |
