摘要
背景:在肿瘤细胞中,β-catenin降解障碍导致胞浆内游离的β-catenin聚集并与TCF/LEF结合进入细胞核内,启动下游基因转录,诱发Wnt/β-catenin信号通路异常改变。因此,研究β-catenin与卵巢癌干细胞的关系,可为特异性抑制卵巢癌干细胞提供新的思路。目的:观察β-catenin基因沉默后卵巢癌干细胞增殖、迁移与侵袭能力的变化。方法:采用免疫磁珠法从人卵巢癌细胞A2780中分离CD133阳性卵巢癌干细胞。选取70%-80%融合的CD133阳性细胞,分别转染慢病毒p LKO.1-sh RNA-β-catenin(实验组)与p LKO.1-sh RNA-ctrl(对照组)质粒,转染后72 h,采用Western blot检测两组细胞β-catenin、上皮表型分子E-cadherin和间质表型分子Vimentin的表达,MTT法检测两组细胞增殖活性,Transwell小室实验检测两组细胞迁移与侵袭能力。结果与结论:(1)转染后72 h,实验组细胞β-catenin蛋白明显低于对照组(P<0.05);(2)实验组培养2,3 d的细胞增殖能力低于对照组(P<0.05);(3)转染后72 h,实验组细胞迁移能力、侵袭能力明显低于对照组(P<0.05);(4)转染后72 h,实验组上皮表型分子E-cadherin蛋白表达高于对照组(P<0.05),间质表型分子Vimentin蛋白表达低于对照组(P<0.05);(5)结果表明,Wnt/β-catenin信号通路参与了卵巢癌干细胞的迁移与侵袭,沉默β-catenin基因表达可抑制卵巢癌干细胞的增殖、迁移及侵袭能力。
BACKGROUND:Inability of degradation makes free cytosolicβ-catenin in tumor cells aggregate and bind to TCF/LEF into the nuclei,which initiates transcription of downstream genes,and eventually induces abnormal changes in Wnt/β-catenin signaling pathways.Therefore,investigations on the relationship betweenβ-catenin and ovarian cancer stem cells can provide a new insight into the specific inhibition of ovarian cancer stem cells.OBJECTIVE:To observe the proliferation,migration and invasion abilities of ovarian cancer stem cells afterβ-catenin gene silencing.METHODS:CD133 positive tumor stem cells were isolated from human ovarian cancer cell line A2780 by immunomagnetic beads.CD133 positive cells with 70%-80%confluence were selected and transfected with pLKO.1-shRNA-β-catenin(experimental group)and pLKO.1-shRNA-ctrln(control group),respectively.The expression ofβ-catenin,E-cadherin and Vimentin were detected by western blot at 72 hours after transfection.Cell proliferation was detected by MTT,and cell migration and invasion ability was detected by Transwell chamber assay.RESULTS AND CONCLUSION:(1)The expression ofβ-catenin protein in the experimental group was significantly lower than that in the control group at 72 hours after transfection(P<0.05).(2)With the extension of culture time,the proliferation ability of the two groups increased gradually,but the cell proliferation ability of the experimental group was lower than that of the control group at 2 and 3 days after transfection(P<0.05).(3)After 72 hours of transfection,the cell migration and invasion abilities of the experimental group were significantly lower than those of the control group(P<0.05).(4)At 72 hours after transfection,the expression of E-cadherin protein in the experimental group was significantly higher than that of the control group(P<0.05),and the expression of Vimentin protein was lower than that of the control group(P<0.05).To conclude,the Wnt/β-catenin signaling pathway is involved in the migration and invasion of ovarian cancer stem
作者
姚彬
张庆华
Yao Bin;Zhang Qing-hua(The Central Hospital of Wuhan,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430000,Hubei Province,China)
出处
《中国组织工程研究》
CAS
北大核心
2018年第25期4001-4006,共6页
Chinese Journal of Tissue Engineering Research
