髓系鋅指基因與神經膠母細胞瘤轉移的機制探討

人類惡性腦瘤細胞具有快速增生以及強烈轉移能力的特性，而惡性腦瘤病人的術後存活率都很低且復發率極高，因此尋找控制惡性腦瘤的標靶因子是現今很重要的課題。而骨髓鋅拇指蛋白-1 (Myeloid zinc finger 1；MZF1) 屬於鋅拇指蛋白中Kruppel家族中的成員之一，其會參與腫瘤細胞的生長、分化與凋亡。近期有文獻指出MZF1會誘導一些腫瘤細胞移動與侵襲能力增加，但是MZF1在腦瘤細胞的生長與轉移能力中所扮演的角色，至今尚未釐清，因此本研究將探討MZF1在臨床檢體上的表現，並進一步利用細胞實驗研究MZF1在腦瘤細胞的分子機轉中所扮演的角色。首先利用免疫組織化學染色法 (Immunohistochemistry；IHC) 偵測腦瘤病人組織臨床切片上MZF1的表現，結果顯示MZF1在膠質母細胞瘤上大量表現；另外統計各分期的腦瘤病人其組織中MZF1的表現，結果發現在低分化性以及第三、第四期癌症病人中，MZF1高表現的數量占較高比例。而因GBM8401人類腦瘤細胞的MZF1表現量相對較高，我們進一步篩選了MZF1基因沉默化的GBM8401人類腦瘤細胞株進行研究，由Boyden chamber assay的結果發現MZF1沉默化會抑制GBM8401細胞的移動和侵襲能力。接著利用反轉錄聚合酶連鎖效應 (reverse transcription-polymerase chain reaction；RT-PCR) 和西方墨點法 (western blot) 的實驗發現MZF1沉默化也會抑制腫瘤轉移相關蛋白-2 (Metastasis tumor-associated protein-2；MTA2) 其蛋白以及mRNA的表現，另外，由於U87-MG人類腦瘤細胞的MZF1表現量相對較低，因此我們也篩選了MZF1大量表現的U87-MG人類腦瘤細胞株，也發現MZF1大量表現會增加MTA2的蛋白及mRNA的表現，並抑制細胞的移動和侵襲能力。由IHC結果推論MZF1的表現可作為預測腦瘤的指標，綜合以上結果，MZF1在未來也許可以成為腦瘤患者治療的目標因子。

Human malignant glioma cells are characterized by invaded-growth of adjacent tissues. The literature indicates that patient’s postoperative survival rate is very low and the recurrence rate is high, thus, searching the target factor of brain cancer is a critical research objective. MZF1 (Myeloid zinc finger 1) is one of the kruppel type zinc finger family, that associated with tumor growth, proliferation and apoptosis. The literature has reported that MZF1 can induce tumor cell movement and invasion. However, the mechanism of MZF1 in brain cancer remains unclear. The purpose of the experiment in the current study was to investigate the correlation between MZF1 and brain cancer. The results indicated that the MZF1 expression in glioblastoma was substantially higher than that of normal tissues via the Immunohistochemistry assay. We also found that the high MZF1 expression number in poorly-differentiated cancer and grade III+IV was bigger than that in well-differentiated cancer and grade I+II. Subsequently, we conducted a Boyden chamber assay to examine GBM8401 human glioma cell line, which consistently silences MZF1. The results indicated that the silence MZF1 decreased the movement and invasion of GBM8401 cells. In addition, we used western blot and reverse transcription-polymerase chain reaction (RT-PCR) and found that MZF1 silence decreased the protein and mRNA expression of MTA2 (Metastasis tumor-associated protein-2). We also used RT-PCR, western blot and Boyden chamber assay to examine U87-MG human glioma cell line, which consistently overexpression MZF1. The results indicated that the overexpression of MZF1 increased the movement and invasion of U87-MG cell, and the mRNA and protein expression of MTA2. Based on the aforementioned results, MZF1 may be the target factor to therapy the brain cancer patients in the future.