胃癌是全世界常見死亡癌症，而亞洲是胃癌高盛行的地區。根據衛生署統計資料顯示，2008年胃癌在台灣十大癌症中排名第五位。目前手術切除腫瘤是胃癌主要治療的方法。早期胃癌五年存活率可達九成，但晚期胃癌五年存活率則低於30%，而胃癌細胞轉移是造成癌症病人死亡的主要原因。因此腫瘤細胞侵入的機轉在控制胃癌的轉移扮演關鍵性的角色。
Osteopontin (OPN)是一個分泌性磷酸化的醣蛋白，許多文獻皆指出，OPN在不同生理與病理機制上具有重要作用。研究顯示OPN可與細胞表面分子integrin或CD44結合而具有促進腫瘤細胞存活、侵入與轉移的能力的多重功能。OPN在各種腫瘤組織如胃癌、乳癌與其他癌症皆呈現高度表達現象，而許多研究也指出高濃度的OPN胃癌的發展進程與轉移密切相關。
人類RUNX3轉錄因子已被證明是一個腫瘤抑制蛋白，在胃上皮細胞正常生長與分化扮演相當重要角色。然而在胃癌病患中常發現RUNX3的啟動子過度甲基化，使RUNX3表現下降。因此，RUNX3功能不活化可能促進胃癌的發生與發展進程。
在本研究中，我們分析OPN啟動子4個RUNX3可能的結合位。結果顯示OPN表達的調控機制中，RUNX3功能為轉錄抑制因子，抑制OPN的轉錄。因此，RUNX3的表現低降，可能導致OPN過度表達，促發腫瘤形成或胃癌細胞的轉移。進一步，我們利用ChIP assay 與 reporter assay，確認在OPN promoter上具有一個RUNX3有效的結合位。將該結合位進行突變後則回復OPN啟動子被RUNX3抑制的轉錄活性。綜合上述結果，我們證明RUNX3可直接結合到OPN的啟動子，作為transrepressor，抑制OPN轉錄的活性。

Gastric cancer is one of the most common causes of cancer-related death throughout the world and has a high prevalence in Asia. In Taiwan, gastric cancer ranked fifth in mortality in 2008. Surgery is the only potentially curative treatment with gastric cancer. The 5-year survival rate for patients diagnosed in the early stages is 90%, while for those in the advanced stages it is only 20-30%. Tumor cell metastasis is the leading cause for gastric cancer mortality. Therefore, the mechanisms of cell motility and invasion are considered to be involving in the tumor progression and metastasis of gastric cancer. The OPN protein is a secreted phosphoglycoprotein involved in a number of cell physiological processes. Studies have suggested that OPN has multifunctional properties in promoting cell survival, cell invasion, and cell migration through interaction with the integrins and CD44 receptors. Elevated expression of OPN has been found in a number of tumors including gastric cancer breast, and others. Furthermore, various outstanding have suggested that high concentration of OPN is highly correlated with gastric cancer progression and metastasis. The human runt-related transcription factor 3 (RUNX3), a tumor suppressor has been suggested to play a vital role in both cell proliferation and differentiation in gastric epithelia. Moreover, the promoter of RUNX3 is hypermethylated has been found in gastric cancer. Therefore, it is reasonable that inactivation of the RUNX3 could facilitate development or progression of gastric cancer. In this present study, we analyze four putative RUNX3 binding sites within the promoter of OPN. Our data suggested that Runx3 is a transcription repressor to suppress OPN expression. Loss of Runx3 leads to overexpression of OPN and promotes tumor progression and metastasis. In addition, a novel potentially RUNX3 binding sites was indentified within OPN promoter by ChIP assay and reporter assay. Mutation of this site can be reversed the transcriptional repressive effect of RUNX3. Altogether, our data demonstrated that RUNX3 acts as a transrepressor directly binding to the promoter of OPN and further suppresses the transcription activity of OPN.

http://www.doh.gov.tw (行政院衛生署)

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