本文選題：Rap2 + 細(xì)胞遷移�。� 參考：《哈爾濱工業(yè)大學(xué)》2014年碩士論文

【摘要】：Rap2屬于Ras家族Rap亞族，具有Rap2A、Rap2B和Rap2C三種異構(gòu)體，調(diào)節(jié)多種細(xì)胞生命活動。由于Rap2與Ras家族其他成員具有相似的結(jié)構(gòu)，因此Rap2能夠結(jié)合多種Ras家族共有的相互作用蛋白，另外Rap2還可能通過與特異性相互作用蛋白的結(jié)合的方式完成信號轉(zhuǎn)導(dǎo)。目前已知的人源Rap2相互作用蛋白有PARG、TNIK、MINK和MAP4K4四種，，但是對其在腫瘤發(fā)生及發(fā)展中的機(jī)制尚無闡述。 本課題組前期研究發(fā)現(xiàn)活性化Rap2在腎癌組織中的水平遠(yuǎn)高于癌旁正常組織，且Rap2的高表達(dá)能夠促進(jìn)細(xì)胞的運(yùn)動和遷移，然而其分子機(jī)制都尚待闡明。本文首先通過GST-Pull down檢測了腎癌細(xì)胞系中Rap2的活性化水平，并通過半定量PCR檢測了對Rap2活性具有調(diào)控作用的Rap1GAP1和RasGEF1B在不同腎癌細(xì)胞系的表達(dá)水平，結(jié)果發(fā)現(xiàn)腎癌細(xì)胞系中Rap2的活性化水平與上述調(diào)控因子的差異表達(dá)相關(guān)。為了研究Rap2調(diào)控細(xì)胞侵襲和遷移的作用通路，通過劃痕實(shí)驗(yàn)和Transwell實(shí)驗(yàn)對具有高活性化水平Rap2的腎癌細(xì)胞系進(jìn)行了針對TNIK和MAP4K4的RNA干擾和遷移及侵襲分析，結(jié)果顯示：Rap2通過TNIK和MAP4K4促進(jìn)腎癌細(xì)胞的遷移及侵襲能力。已有研究報道FAK正調(diào)控細(xì)胞的鋪展和運(yùn)動，本文通過免疫熒光和western blot的實(shí)驗(yàn)證明Rap2及其激活突變體的過表達(dá)可上調(diào)FAK的表達(dá)，而DN突變體Rap2-17N則對FAK表達(dá)無顯著調(diào)控作用。本研究增進(jìn)了對于Rap2在腎癌細(xì)胞系中高活性化的機(jī)制，以及Rap2調(diào)控細(xì)胞運(yùn)動和侵襲的分子機(jī)制的了解。
[Abstract]:Rap2 belongs to the Rap subfamily of Ras family. It has three isomers Rap2An Rap2B and Rap2C and regulates many cell life activities. Because Rap2 has similar structure with other members of Ras family, Rap2 can bind to many kinds of interaction proteins shared by Ras family, and Rap2 may also complete signal transduction by binding with specific interacting proteins. At present, there are four known human Rap2 interaction proteins: Rap2 TNIKMIK and MAP4K4, but the mechanism of them in tumorigenesis and development has not been elucidated. Our previous study found that the level of activated Rap2 in renal cell carcinoma tissues is much higher than that in adjacent normal tissues, and the high expression of Rap2 can promote cell movement and migration, but its molecular mechanism remains to be clarified. In this study, the activation level of Rap2 was detected by GST-Pull down, and the expression of Rap1GAP1 and RasGEF1B, which regulate the activity of Rap2, was detected by semi-quantitative PCR in different RCC cell lines. The results showed that the activation level of Rap2 was related to the differential expression of these regulatory factors in renal cell carcinoma cell line. In order to study the pathway of Rap2 regulating cell invasion and migration, the RNA interference, migration and invasion of TNIK and MAP4K4 in renal cell lines with high Rap2 activity were analyzed by scratch test and Transwell assay. The results showed that: Rap2 promoted the migration and invasion of renal cancer cells through TNIK and MAP4K4. It has been reported that FAK is regulating the proliferation and movement of cells. The results of immunofluorescence and western blot have shown that the overexpression of Rap2 and its activated mutants can up-regulate the expression of FAK, while the Rap2-17N of DN has no significant effect on FAK expression. This study enhanced the understanding of the mechanism of high activation of Rap2 in renal cell line and the molecular mechanism of Rap2 regulating cell movement and invasion.
【學(xué)位授予單位】：哈爾濱工業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R737.11

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 蔣新農(nóng),周柔麗;腫瘤細(xì)胞粘附、遷移與轉(zhuǎn)移的相關(guān)性[J];生物化學(xué)與生物物理進(jìn)展;1998年05期

本文編號：1800014