摘要
目的:探讨β-catenin蛋白对卵巢癌干细胞特性及细胞增殖的影响。方法:采用β-catenin siRNA技术沉默SKOV3中β-catenin基因,形成SKOV3-RNAi细胞。转染空载体siRNA的SKOV3-NC细胞作为对照。流式细胞技术测定CD44、CD117表达;通过卡铂作用后细胞的平板克隆计数及MTT法检测沉默β-catenin基因后细胞增殖能力,通过裸鼠背部皮下移植瘤实验检测细胞的体内成瘤能力。结果:与SKOV3-NC细胞相比,SKOV3-RNAi细胞在无血清培养条件下细胞球形成明显减少(P<0.05),且CD44、CD117荧光强度均明显降低(P<0.05)。卡铂作用于细胞后,SKOV3-RNAi细胞克隆数的减少量比SKOV3-NC细胞更为显著,同时细胞生长速度明显减慢;裸鼠体内,SKOV3-RNAi的成瘤能力较SKOV3-NC细胞明显降低。结论:沉默β-catenin基因后,卵巢癌细胞系SKOV3的干细胞特性明显减弱,细胞增殖也受到抑制。
Objective： To explore the effect of the expression of β-catenin to ovarian cancer stem cell properties and cell proliferation. Methods： By β-catenin siRNA technology theβ-catenin gene of ovarian cancer cell line SKOV3 was knocked down,which named SKOV3-RNAi cells. The control group SKOV3 cells were transfected with control empty vector siRNA,which named SKOV3-NC cells. Both of these two groups of cells were cultured in serum-free suspension culture. The expressions of CD44 and CD117 were measured by flow cytometry,which was used to analyze the effect of silencing β-catenin gene of SKOV3 to the cancer stem cell properties of ovarian cancer cells. Plate cloning experiments,was used to count the number of cell formation after disposed by carboplatin. Detecting the cell growth by MTT method can reflect the effect of silence β-catenin gene to cell proliferation. In vivo,the proliferation of the cells was detected by subcutaneous transplantation tumor in nude mice. Results： Compared with the control group SKOV3-NC cells,the cell spheres of SKOV3-RNAi cells in serum-free culture conditions were significantly reduced（ P 〈0. 05）,and the flow cytometry found the fluorescence intensity of CD44,CD117 of SKOV3-RNAi cell were lower than SKOV3-NC cells（ P〈 0. 05）. The plates clone assay showed that,after disposed to carboplatin,the proliferation of SKOV3-RNAi cells decreased more than SKOV3-NC cells as the number of cell colony formation was significant reduced. MTT assay found that,compared with the SKOV3-NC cells,the growth rate of SKOV3-RNAi cells slowed down. In vivo,SKOV3-RNAi tumorigenic ability much more decreased than SKOV3-NC cells significantly. Conclusion： Silence the β-catenin gene of ovarian cancer cell line SKOV3 decreased the characteristics of stem cells significantly,and the cell proliferation is also inhibited.
出处
《现代妇产科进展》
CSCD
北大核心
2016年第8期571-574,577,共5页
Progress in Obstetrics and Gynecology
基金
河南省医学科技攻关计划(No:201301007)
