卵巢癌SKOV3细胞中筛选获得的全克隆细胞的生物学特性及其形成血管生成拟态的能力

发布时间：2018-08-12 11:06

【摘要】：目的 :观察从卵巢癌SKOV3细胞中筛选获得的全克隆(holoclone)细胞的特性及其形成血管生成拟态(vasculogenic mimicry,VM)的能力。方法 :通过有限稀释法对SKOV3细胞进行单细胞培养,形成3种单细胞克隆[全克隆、部分克隆(meroclone)和亚克隆(paraclone)];通过裸鼠体内成瘤实验和软琼脂克隆形成实验比较3种单克隆的生物学特性,并通过三维细胞培养模型观察其形成VM的能力。蛋白质印迹法检测全克隆细胞中CD133、基质金属蛋白酶2(matrix metalloproteinases-2,MMP-2)及MMP-9的表达水平。结果 :SKOV3细胞单细胞培养10~14 d后,全克隆、部分克隆和亚克隆细胞所占的比例分别为(17.4±0.8)%、(30.1±0.4)%和(52.5±1.1)%。裸鼠体内成瘤率结果显示,全克隆细胞的成瘤率为80%,部分克隆和亚克隆细胞在裸鼠中均未有肿瘤形成。软琼脂克隆形成实验结果提示,全克隆细胞的克隆形成率为(16.7±3.8)%,显著高于部分克隆及亚克隆细胞的(6.5±1.5)%和(1.2±0.7)%(F=4.085,P0.05)。三维培养中,全克隆组可以形成VM,部分克隆和亚克隆细胞不能形成VM。全克隆细胞中CD133、MMP-2和MMP-9蛋白的表达水平均高于亲本SKOV3细胞(P值均0.05),并高于部分克隆和亚克隆细胞(P值均0.05)。结论 :SKOV3细胞中单细胞克隆在形态和生物学特性上存在异质性,其中全克隆细胞亚群具有肿瘤干细胞样细胞特性,具有形成VM的能力。全克隆是SKOV3细胞中具有较高复发能力的细胞亚群。
[Abstract]:Aim: to observe the characteristics of fully cloned (holoclone) cells isolated from ovarian cancer SKOV3 cells and their ability to form vasculogenic mimicry VM. Methods: SKOV3 cells were cultured by finite dilution method to form three kinds of single cell clones. Partial cloning of (meroclone) and subcloning of (paraclone)], the biological characteristics of three kinds of monoclonal clones were compared by tumorigenesis in nude mice and soft Agar clone formation, and the ability of VM formation was observed by three-dimensional cell culture model. The expression of CD133, matrix metalloproteinases-2 (MMP-2) and MMP-9 were detected by Western blot. Results after 10 ~ 14 days of single cell culture, the percentage of cloned, partially cloned and subcloned cells were (17.4 卤0.8)%, (30.1 卤0.4)% and (52.5 卤1.1)%, respectively. The results of tumorigenesis in nude mice showed that the tumorigenic rate of all cloned cells was 80%, and no tumor formation was found in some cloned and subcloned cells in nude mice. The results of soft Agar clone formation test showed that the clone formation rate of the whole cloned cells was (16.7 卤3.8), significantly higher than that of the partial and subcloned cells (6.5 卤1.5)% and (1.2 卤0.7)% (P 0.05). In three dimensional culture, the whole clone group could form VM, and some cloned and subcloned cells could not form VM. The expression levels of CD133mMMP-2 and MMP-9 protein in the whole cloned cells were higher than those in the parental SKOV3 cells (P < 0. 05), and were higher than those in some cloned and subcloned cells (P < 0. 05). Conclusion there is heterogeneity in morphology and biological characteristics of single cell clones in SKOV3 cells, in which the whole clone subsets have the characteristics of tumor stem cell like cells and have the ability to form VM. Full cloning is a subgroup of SKOV3 cells with high relapse ability.
【作者单位】：解放军白求恩国际和平医院妇产科;
【分类号】：R737.31

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