白杨素调控miR-320和Twist1表达抑制卵巢癌干细胞样细胞体外致瘤功能

Chrysin inhibits in vitro tumorigenicity of ovarian cancer stem-like cells through modulating expressions of miR-320 and Twist1

目的 :研究白杨素能否调节miR-320和Twist1表达抑制人卵巢癌SKOV3源性干细胞样细胞体外致瘤能力。方法 :不同浓度白杨素(5.0、10.0和20.0μM)处理肿瘤球培养得到的第2代球细胞即卵巢癌干细胞样细胞。实时定量PCR检测miR-320表达水平。球形成率测定和琼脂集落形成试验评价体外致瘤能力。Western blot分析Twist1蛋白表达。分别用miR-320模拟物或Twist1基因转导探讨白杨素的作用机制。结果 :与亲本细胞相比,SKOV3源性卵巢癌干细胞样细胞miR-320低表达,而Twist1蛋白高表达。白杨素(5、10和20μM)以浓度依赖方式降低SKOV3源性卵巢癌干细胞样细胞球形成率和集落形成率;同时,上调miR-320表达和下调Twist1蛋白表达。miR-320模拟物转染增强白杨素(10μM)的效应。Twist1基因转导能有效逆转白杨素(10μM)的作用。结论 :白杨素通过调控miR-320/Twist1信号轴抑制SKOV3源性卵巢癌干细胞样细胞体外致瘤性。

Objective To investigate whether chrysin can inhibit in vitro tumorigenicity of ovarian cancer stem-like cells(OCSLCs) derived from human ovarian cancer SKOV3 cell line by modulating the expression of Mi R-320 and Twist1. MethodsSKOV3-derived OCSLCs were obtained from the second generation of cancer cells using tumor spheroid formation culture, which were treated with various concentrations(5.0, 10.0 and 20.0μM) of chrysin. Real-time quantitative PCR was used to detect the expression of Mi R-320. Ttumorigenicity in vitro were evaluated by Spheroid and colony formation assay. Western blot was used to examine Twist1 protein expression. Mi R-320 mimics transfection or Twist1 gene transduction were used to elucidate the mechanism underlying the action of chrysin in OCSLCs. Results The miR-320 of SKOV3-derived OCSLCs was low-expressed and Twist1 protein was highly expressed, compared to the parental cells. Chrysin(5, 10 and 20μM) decreased spheroid and colony forming rates of SKOV3-derived OCSLCs in a concentration-dependent manner, while up-regulated the expression of miR-320 and down-regulated Twist1. miR-320 mimics transfection enhances the effect of chrysin(10μM). Twist1 gene transduction can effectively reverse the effect of chrysin(10μM). Conclusion Chrysin inhibits in vitro tumorigenicity of SKOV3-derived OCSLCs by regulating the miR-320/Twist1 axis.