白杨素对SKOV3细胞系干样细胞球形成和CK2α仪表达的影响

Effects of chrysin on sphere formation and CK2α expression of ovarian cancer stem-like cells derived from SKOV3 cell line

目的 探讨白杨素抑制卵巢癌干样细胞（人卵巢癌SKOV3细胞系球细胞）球形成作用是否涉及下调酪蛋白激酶催化亚单位-2α（CK2α）蛋白表达.方法 不同浓度白杨素（5.0、10.0和20.0 μmol/L）处理干细胞培养基超低黏附板悬浮培养得到的卵巢癌干样细胞.球形成法测定细胞球形成率.免疫印迹分析细胞CK2α和干细胞标志物CD133、CD44蛋白表达水平.CK2α siRNA与pcDNA3.1-CK2α质粒转染探讨白杨素的作用机制.结果 白杨素（5.0、10.0和20.0 μmol/L）降低卵巢癌干样细胞球形成率（22.3％±2.5％比14.7％ ±2.1％、8.6％±1.7％比3.8％±1.1％;P＜0.05）,并下调CK2α、CD133和CD44蛋白表达.CK2α siRNA转染增强白杨素作用.pcDNA3.1-CK2α质粒转染减弱白杨素作用.结论 下调CK2α蛋白表达参与白杨素抑制卵巢癌干样细胞球形成作用.

Objective To investigate whether inhibitory effect of chrysin on sphere formation of ovarian cancer stem-like cells（spheroids derived from human ovarian cancer SKOV3 cell line） is involved in the down-regulating of the protein expression of casein kinase CK2α.Methods SKOV3-derived ovarian cancer stem-like cells obtained by suspension culture in stem cell-condition medium using ultra-low adhesion plate were treated with various concentrations （5.0,10.0 and 20.0 μmol/L） of chrysin.Sphere formation assay was used to determine the sphere forming rate of SKOV3-derived ovarian cancer stem-like cells.Western blot was used to analyze the protein expressions of CK2α and cancer stem cell markers CD133 and CD44.Silence of CK2α by siRNA and ectopic expression of CK2α by transfection with pcDNA3.1-CK2α plasmid were used to explore the mechanism underlying the effect of chrysin on sphere formation of SKOV3-derived ovarian cancer stem-like cells.Results Chrysin （5.0,10.0 and 20.0 μmol/L） significantly reduced the sphere forming rate of SKOV3-derived ovarian cancer stem-like cells,in a concentrationdependent manner （22.3% ±2.5% vs 14.7% ±2.1%,8.6% ± 1.7% and 3.8% ± 1.1% respectively;P 〈0.05）.In addition,chrysin （5.0,10.0 and 20.0 μmol/L） obviously down-regulated the protein expressions of CK2α,CD133 and CD44 in SKOV3-derived ovarian cancer stem-like cells.In combination with CK2α siRNA transfection and chrysin synergistically decreased sphere formation （P 〈 0.05） and the protein expressions of CK2α,CD133 and CD44 in SKOV3-derived ovarian cancer stem-like cells.However,transfection with pcDNA3.1-CK2α plasmid attenuated inhibitory effects of chrysin on sphere formation capability and the expressions of CK2α,CD133 and CD44 of SKOV3-derived ovarian cancer stem-like cells.Conclusion Down-regulation of CK2α protein expression is involved in the inhibition effect of chrysin on the sphere formation capability of SKOV3-derived ovarian cancer stem-like cells.