本文選題：IQGAP1 + 卵巢癌��； 參考：《浙江大學(xué)》2014年碩士論文

【摘要】：目的： 探索支架蛋白IQGAP1在卵巢癌腫瘤干細(xì)胞的表達(dá),并觀察IQGAP1對(duì)卵巢癌腫瘤干細(xì)胞分化過(guò)程中的侵襲能力的影響。 方法： 以漿液性卵巢癌3AO細(xì)胞株為親本細(xì)胞,無(wú)血清懸浮培養(yǎng)法獲得腫瘤干細(xì)胞。采用流式細(xì)胞術(shù)驗(yàn)證腫瘤干細(xì)胞的標(biāo)記分子CD24表達(dá),利用細(xì)胞免疫熒光實(shí)驗(yàn)觀察干性分子OCT4和SOX2的定位。利用含10%胎牛血清培養(yǎng)基誘導(dǎo)其分化,期間光鏡下觀察細(xì)胞形態(tài)變化。 qRT-PCR法和Western-blot法分別檢測(cè)IQGAP1mRNA和蛋白的表達(dá)水平變化。通過(guò)劃痕實(shí)驗(yàn)和Transwell小室觀察細(xì)胞遷移侵襲能力；采用wst-1法檢測(cè)細(xì)胞增殖能力。 結(jié)果： 1.利用3AO細(xì)胞株通過(guò)無(wú)血清培養(yǎng)法獲得表型CD24(-)表型的腫瘤干細(xì)胞。 2.相比3AO細(xì)胞株中,IQGAP1在腫瘤干細(xì)胞中呈低表達(dá)。 3.在3AO細(xì)胞株中,沉默IQGAP1后減弱了腫瘤遷移、侵襲能力,而不會(huì)改變細(xì)胞的增殖活性。 4.在卵巢腫瘤干細(xì)胞分化過(guò)程中,OCT4和SOX2表達(dá)下降,而IQGAP1表達(dá)增加；同時(shí)腫瘤細(xì)胞的侵襲能力增強(qiáng)。 5.在卵巢腫瘤干細(xì)胞分化過(guò)程中,IQGAP1沉默后細(xì)胞侵襲能力減弱。 結(jié)論： 1.無(wú)血清培養(yǎng)法是獲得腫瘤干細(xì)胞比較成熟簡(jiǎn)便的實(shí)驗(yàn)技術(shù)。 2.在腫瘤干細(xì)胞分化過(guò)程中其侵襲能力逐步增強(qiáng),且IQGAP1可能參與了該變化。
[Abstract]:Objective:
Objective to explore the expression of scaffold protein IQGAP1 in ovarian cancer stem cells and observe the effect of IQGAP1 on invasion of ovarian cancer stem cells.
Method:
A serous ovarian cancer 3AO cell line was used as a parent cell to obtain tumor stem cells without serum suspension culture. Flow cytometry was used to verify the expression of marker molecule CD24 in tumor stem cells. The localization of OCT4 and SOX2 of dry molecules was observed by cell immunofluorescence test. The differentiation was induced by 10% fetal bovine serum medium. The morphological changes of the cells were observed.
The changes of expression level of IQGAP1mRNA and protein were detected by qRT-PCR and Western-blot. The proliferation and invasion ability of cells were observed by scratch test and Transwell compartment, and the proliferation ability of cells was detected by WST-1.
Result錛，

本文編號(hào)：2055360