探討間質蛋白酶在攝護腺癌細胞中對細胞基質第九金屬蛋白酶之影響

Matriptase induced the expression and activation of matrix metalloproteinase 9 (MMP-9) in human prostate cancer PC-3 cells

王欣韻(Hsin-Yun Wang)

指導教授 ： 李明學

國立臺灣大學/醫學院/生物化學暨分子生物學研究所/碩士(2012年)

https://doi.org/10.6342/NTU.2012.01636

摘要

攝護腺癌在台灣有逐年提升的趨勢，高居男性癌症死亡率中的第七位。一般來說，癌細胞轉移是造成癌症病人死亡的主要原因。癌細胞的轉移過程中，常伴隨著蛋白酶活性的提升，進而促進細胞間質的降解，使得癌細胞得以轉移至其它組織生長。近年來的研究指出，屬於第二型嵌膜絲胺酸蛋白酶的間質蛋白酶，參與各種癌症的惡化過程。然而，間質蛋白酶如何促進攝護腺癌細胞侵襲及惡化的分子機制目前仍未清楚。為了釐清間質蛋白酶在攝護腺癌細胞惡化中的角色，我們建立了具高侵襲能力的攝護腺癌細胞模式，並發現間質蛋白酶及基質金屬第九蛋白酶的表現量會隨著細胞的侵襲能力提升而隨之增加。為了進一步探討間質蛋白酶是否對基質金屬第九蛋白酶的表現或活性有所調控，我們在 PC-3 細胞中，利用質體轉殖方式送入間質蛋白酶使其表現增加，我們研究觀察到基質金屬第九蛋白酶表現量及其活性，會隨著間質蛋白酶活性態提高而增加。且利用單點突變方式證明，間質蛋白酶的活性對於基質金屬第九蛋白酶的調控十分重要。此外，利用核酸干擾技術壓抑間質蛋白酶的表現後，也觀察到基質金屬第九蛋白酶的活性減少。整體來說，在癌症惡化過程中，間質蛋白酶的增加，可能進一步經由活化基質金屬第九蛋白酶，以促進癌細胞的轉移惡化。

關鍵字

攝護腺癌 ； 間質蛋白酶基質金屬第九蛋白酶癌症惡化 ； 細胞運動

並列摘要

The main cause of prostate cancer patient death is due to cancer metastasis. During metastasis, invading cells often up-regulate pericellular proteases to degrade extracellular matrix (ECM). Matriptase, a membrane-anchored serine protease, is thought to be involved in tumor progression. However, the detailed molecular mechanisms in which matriptase promotes prostate cancer progression and invasion are still unclear. To clarify the roles of matriptase in prostate cancer progression, we established a prostate cancer PC-3 cell progression model by serially isolating highly invasive cancer cells using matrigel-coated Boyden chambers. In this progression model, the expression and activated levels of matriptase and matrix metalloproteinase 9 (MMP-9) were increased following the cancer progression and correlated with cancer cell invasion. To further analyze whether matriptase can induce the expression or gelatinolytic activity of MMP-9, we performed overexpression experiments to increase the activated level of matripase in PC-3 cells and found that matriptase overexpression induced MMP-9 gelatinolytic activity and expression, while a catalytically inactive mutant of matriptase (S805A) had no significant effect on the biological events. Moreover, matriptase knockdown by siRNA to down-regulate MMP-9 activity. Taken together, the data indicated that these increase levels of activated matriptase following the progression of prostate cancer cells was involved in up-regulating MMP-9 and promoting cancer cell invasion.

並列關鍵字

prostate cancer ； matriptase ； matrix metalloproteinase 9 ； cancer progression ； cell motility

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