卵巢癌肿瘤干细胞分化过程中支架蛋白IQGAP1变化的体外研究

发布时间：2018-06-23 02:34

  本文选题：IQGAP1 + 卵巢癌　； 参考：《浙江大学》2014年硕士论文

【摘要】：目的： 探索支架蛋白IQGAP1在卵巢癌肿瘤干细胞的表达,并观察IQGAP1对卵巢癌肿瘤干细胞分化过程中的侵袭能力的影响。 方法： 以浆液性卵巢癌3AO细胞株为亲本细胞,无血清悬浮培养法获得肿瘤干细胞。采用流式细胞术验证肿瘤干细胞的标记分子CD24表达,利用细胞免疫荧光实验观察干性分子OCT4和SOX2的定位。利用含10%胎牛血清培养基诱导其分化,期间光镜下观察细胞形态变化。 qRT-PCR法和Western-blot法分别检测IQGAP1mRNA和蛋白的表达水平变化。通过划痕实验和Transwell小室观察细胞迁移侵袭能力；采用wst-1法检测细胞增殖能力。 结果： 1.利用3AO细胞株通过无血清培养法获得表型CD24(-)表型的肿瘤干细胞。 2.相比3AO细胞株中,IQGAP1在肿瘤干细胞中呈低表达。 3.在3AO细胞株中,沉默IQGAP1后减弱了肿瘤迁移、侵袭能力,而不会改变细胞的增殖活性。 4.在卵巢肿瘤干细胞分化过程中,OCT4和SOX2表达下降,而IQGAP1表达增加；同时肿瘤细胞的侵袭能力增强。 5.在卵巢肿瘤干细胞分化过程中,IQGAP1沉默后细胞侵袭能力减弱。 结论： 1.无血清培养法是获得肿瘤干细胞比较成熟简便的实验技术。 2.在肿瘤干细胞分化过程中其侵袭能力逐步增强,且IQGAP1可能参与了该变化。
[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锛，

本文编号：2055360