第十號程序性細胞死亡分子對於腦癌轉移之抑制現象與機制研究

The Inhibition and Mechanism of Programmed Cell Death 10 (PDCD10) on Brain Cancer Cell Metastasis

林子為(Tz-Wei Lin)

指導教授 ： 黎耀基 ； 張文祥

國立清華大學/生命科學院/生物科技研究所/碩士(2007年)

https://doi.org/10.6843/NTHU.2007.00542

摘要

第十號程序性細胞死亡分子 (programmed cell death 10, PDCD10, TFAR15) 屬於細胞凋亡相關基因家族成員之一。PDCD10首次被報導是當培養人類骨髓癌細胞株TF-1時，在缺乏粒細胞巨噬細胞集落刺激因子(granulocyte macrophage colony- stimulating factor, GM-CSF) 的狀態下，細胞會走向凋亡而造成PDCD10的基因表現量被提高。根據病理統計報導，PDCD10基因突變與顱內海綿狀血管瘤的發生率 (cerebral cavernous malformation, CCM) 極具正相關性，故此基因又被稱為CCM3 (cerebral cavernous malformation 3) 基因。根據研究報導指出，在培養人類急性骨髓白血病細胞株HL-60時，若缺乏胎牛血清的狀態下，PDCD10的基因表現量亦會提高，並可透過細胞外信號調節激酶 (extracellular signal-regulated kinase, ERK) 來促進細胞增生以及細胞轉化的現象。意外的是，根據本論文的研究，在缺乏胎牛血清的細胞培養狀態下，HL-60細胞的PDCD10基因表現量反倒呈現著下調的情形。此外，根據純化之第十號程序性細胞死亡分子重組蛋白，我們成功製備了PDCD10的多株抗體。利用此抗體，我們進一步証明了HL-60細胞在缺乏胎牛血清的培養狀態下，PDCD10的蛋白質表現量ㄧ樣是呈現著下調的情形。而另一方面當大量表現PDCD10蛋白質於神經膠質瘤細胞株U-87 MG時，可以發現到癌細胞的轉移能力受到明顯抑制，並且基質金屬蛋白酶9 (matrix metalloproteinase 9, MMP9) 的表現量與ERK的活性被上調增加。利用絲裂原活化蛋白激酶的激酶 (mitogen-activated protein kinase kinase, MAPKK) 抑制劑PD98059來抑制細胞內的ERK被活化時，可以觀察到MMP9的表現量減少。因此證明了PDCD10極有可能具有抑制細胞凋亡的功能，並藉由ERK來調控MMP9的表現量進而影響著細胞的侵襲轉移或凋亡。

關鍵字

第十號程序性細胞死亡分子 ； 腦癌轉移 ； 細胞凋亡 ； 細胞外信號調節激酶 ； 基質金屬蛋白酶9

並列摘要

Programmed cell death 10 (PDCD10, TFAR15) is a member of the PDCD family associated with cell apoptosis. It was originally identified in the human premyeloid TF-1 cells for being up-regulated, in which apoptosis was induced by withdrawal of granulocyte macrophage colony-stimulating factor (GM-CSF). Clinical statistics have shown that mutations of PDCD10 are correlated with cerebral cavernous malformation (CCM) and thus this gene is also named as CCM3 (cerebral cavernous malformation 3) accordingly. The expression of PDCD10 was reported to be up-regulated in human promyeloblast HL-60 cells after serum starvation. The elevated PDCD10 then modulates the ERK (extracellular signal-regulated kinase) signaling pathway to promote cell growth and cellular transformation. Surprisingly, in this thesis it is found that the PDCD10 expression is in fact down-regulated in serum-free culture condition. By using the purified PDCD10 recombinant protein as an antigen, I successfully generated mouse anti-human PDCD10 polyclonal antibody. With this new antibody, I was able to further clarify that the PDCD10 protein expression is down-regulated in HL-60 cells after serum starvation. Meanwhile, overexpression of PDCD10 not only significantly suppresses the invasive ability of human glioblastoma U-87 MG cells, but also increases the expression and secretion of MMP9 as well as the ERK activity. Treatment of mitogen-activated protein kinase kinase (MAPKK) inhibitor PD98059 resulted in a suppression of the ERK activity and subsequent decrease of the MMP9 expression. Together these results suggest that PDCD10 may be an anti-apoptotic gene which affects cell apoptotic process as well as tumor cell invasion and metastasis by regulating the expression of MMP9 via the ERK pathway.

並列關鍵字

Programmed Cell Death 10 ； Brain Cancer Cell Metastasis ； Apoptosis ； extracellular signal-regulated kinase ； matrix metalloproteinase 9

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