探討Haspin在人類大腸癌腫瘤形成之角色

Abstract

大腸癌是台灣及世界成為發生率及致死率最高的癌症之一。Haspin是一種Histone 3的磷酸化激酶，具有調控細胞有絲分裂進行的作用。然而，Haspin在大腸癌中的功能及是否能成為標靶治療仍不清楚。在本研究中，我們發現在人類大腸癌細胞株HCT116, RKO, HT-29中會有高表現Haspin蛋白，正常大腸細胞FHC表現量較低。下游的蛋白Survivin與H3T3則會在人類大腸癌細胞中大量表現，正常大腸細胞中沒有。過量表現Haspin蛋白後會增加下游H3T3及Survivin的蛋白表現。沈默Haspin後，會減少下游H3T3及Survivin的蛋白表現。我們蒐集26個來自患者的正常大腸組織及腫瘤組織，利用免疫組織化學染色後，發現Haspin在大部分腫瘤組織中有較高的表現。此外，H3T3與Survivin也會在腫瘤組織中大量表現。然而，利用3D立體培養大腸癌病人的腫瘤組織細胞，並利用裸鼠動物模式異體移病人腫瘤模式，研究腫瘤的形成作用。發現在患者的腫瘤組織中會有Haspin的表現。在本論文中，我們證實Haspin在大腸癌細胞及病人的腫瘤組織中有較高的表現，在大腸癌中Haspin會活化H3T3和Survivin並增加細胞的存活幫助腫瘤形成。我們推測Haspin是一個具有潛力的癌症標靶，可作為未來大腸癌治療的新策略。

Colorectal cancer (CRC) has become one of the highest rates of incidence and mortality in Taiwan and the world. Haspin is a Histone 3 phosphorylated kinase that promotes mitotic progression. However, the role of Haspin in CRC and whether the function can apply to be a treatment target is still unclear. In this study, we found Haspin proteins that were expressed in various human CRC cell lines, including HCT116, RKO and HT-29. FHC is a normal colon cell line that relatively expressed low level of Haspin proteins. The downstream protein levels of Survivin and H3T3 were also highly expressed in various CRC cells but not in the normal CRC cells. Overexpression of Haspin proteins increased the protein levels of H3T3 and Survivin. Blockage of Haspin expression reduced the downstream protein levels of H3T3 and Survivin. We collected 26 cases of colorectal normal tissues and tumors of CRC patients. Most cases of CRC tumors were highly expressed Haspin protein levels by immunohistochemistry staining. Besides, the H3T3 and Survivin proteins were also highly expressed in CRC tumors. Moreover, we could use the 3D-tissue culture of CRC tumor growth and the xenografted CRC tumors in nude mice models to observe the tumorigenesis of CRC in patients. The location and expression of Haspin proteins can be detected in the patient CRC tissues. In this thesis, we demonstrate Haspin that is highly expressed in the CRC cells and tumor tissues of clinical patients. The function of Haspin activates the expression of H3T3 and Survivin in increasing the cell survival and tumorigenesis of CRC. Our finding suggest that Haspin is potential cancer target as a novel strategy for CRC therapy.