探討人類MOB2蛋白參與人類纖維肉瘤細胞爬行的機制

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Please use this identifier to cite or link to this item: http://140.128.103.80:8080/handle/310901/25144

Title:	探討人類MOB2蛋白參與人類纖維肉瘤細胞爬行的機制
Other Titles:	Human MOB2 Participates in Migration of Fibrosarcoma Cells
Authors:	劉怡青 Yi Ching Liu
Contributors:	范聖興 Seng Sheen Fan 生命科學系
Keywords:	細胞爬行 cell migration;mob2
Date:	2014
Issue Date:	2015-03-06T08:30:10Z (UTC)
Abstract:	Mps One Binder (MOB)是一個很重要的蛋白來控制細胞內的一些機制，例如細胞增生、有絲分裂、細胞爬行、細胞的型態和死亡。MOB 蛋白家族在酵母菌、果蠅和哺乳動物都可以被找到。先前研究發現MOB蛋白對酵母菌的初芽和分裂是必需的蛋白。在果蠅中，MOB蛋白扮演著腫瘤抑制者來控制Hippo途徑和調控果蠅眼睛的感光接受器的排列。在哺乳類動物中，人類MOB蛋白家族有六種，其中hMOB2被知道是會與NDR1/2磷酸?結核病參與調控細胞凋亡和中心體複製。在類神經細胞中，大量表達MOB2蛋白會增加神經生。除此之外，在星形膠質細胞中，MOB2基因會藉由cAMP-dependent路徑來調控細胞的爬行。細胞爬行在發育、傷口癒合、癌症轉移是一個重要的過程，可以控制細胞在前端的突出以及細胞往前爬時細胞尾端的收縮。在過去的研究中，MOB2對於細胞爬行的機制並不是很清楚，為了研究hMOB2是否參與調控細胞的爬行，我們利用人類纖維腫瘤細胞(HT1080)來做實驗的模型來分析hMOB2是否參與調控細胞爬行。在我的實驗結果可以發現，hMOB2蛋白會分布在爬行細胞的細胞邊緣以及細胞質中。在傷口癒合的實驗和細胞爬行分析中，我們發現減少hMOB2的表達導致細胞的爬行速度變慢。我們也發現再度將hMOB2表達進MOB2 shRNA的細胞中，細胞的爬行功能會恢復。利用A107G, Y110A點突變的hMOB2來測試hMOB2對於細胞的爬行是否是重要的，發現其細胞爬行速率與hMOB2 knockdown細胞相比，並無明顯差異，但速率還是略高於hMOB2 knockdown細胞。我的實驗結果證明hMOB2蛋白在人類纖維肉瘤細胞爬行和入侵中扮演著很重要的功能。 Mps One Binder proteins (MOB) are important components to control cellular processes, such as cell proliferation, cell migration, morphogenesis, and apoptosis. MOB family proteins can be found from yeast, Drosophila and mammals. In Drosophila, Mob1 protein acted as tumor suppressor to control the Hippo pathway and photoreceptor morphogenesis. In mammals, the human MOB protein family has six distinct members. Human (hMOB2) is known to bind with NDR1/2 and participates in regulating cell apoptosis and centrosome duplication. In mouse neuron cell, over-expression of MOB2 increases neurite formation. Cell migration is an important cellular process that occurs during embryo development, wound healing, inflammation and cancer metastasis. Cell migration involves cell protrusion at the leading edge and cell retraction at the rear end of cells. Although the mechanisms of cell migration have been widely studied, the regulation of cell migration remained unclear. To study whether hMOB2 participates in cell migration, we used human fibrosarcoma cell (HT1080) as a model system to study how hMOB2 participates in cell migration. Immunocytochemistry revealed that hMOB2 protein was mainly localized at the cytoplasm and at the leading edge of HT1080 cells. The localization of hMOB2 detected at the leading edge was colocalized with actin cytoskeleton at the lamellipodia. In wound healing and cell invasion assay, we found that the rate of cell invasion was reduced to 50% after knockdown of hMOB2. The invasion rate was restored as hMOB2 proetin was re-expressed in the hMOB2 knockdown cells. We also found that the rate of cell migration was slower in the hMOB2-knockdown cells. In addition, the expression of A107G, Y110A point mutation of hMOB2 in the hMOB2 knockdown cells showed no significant difference between hMOB2 knockdown cells in the rate of cell invasion and cell migration. These results suggest that hMOB2 plays an important function in cell invasion and cell migration.
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