Li H Z, Chen Y H, Fang Y L, Zhong L Y, Yuan Q Q, Xu X Y, Cao J G
Department of Obstetrics and Gynecology, the Fourth Affiliated Hospital of Guangzhou Medical University, Guangzhou 511447, China.
Zhonghua Yi Xue Za Zhi. 2016 Jul 5;96(25):2013-6. doi: 10.3760/cma.j.issn.0376-2491.2016.25.012.
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.
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.
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 concentration-dependent 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.
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.
研究白杨素对卵巢癌干细胞样细胞（源自人卵巢癌SKOV3细胞系的球体）成球的抑制作用是否与酪蛋白激酶CK2α蛋白表达下调有关。
使用超低吸附板在干细胞条件培养基中通过悬浮培养获得源自SKOV3的卵巢癌干细胞样细胞，用不同浓度（5.0、10.0和20.0 μmol/L）的白杨素处理。采用成球实验测定源自SKOV3的卵巢癌干细胞样细胞的成球率。用蛋白质印迹法分析CK2α、癌症干细胞标志物CD133和CD44的蛋白表达。用小干扰RNA（siRNA）沉默CK2α以及用pcDNA3.1-CK2α质粒转染异位表达CK2α，以探讨白杨素对源自SKOV3的卵巢癌干细胞样细胞成球作用的机制。
白杨素（5.0、10.0和20.0 μmol/L）以浓度依赖性方式显著降低源自SKOV3的卵巢癌干细胞样细胞的成球率（分别为22.3%±2.5% 对14.7%±2.1%、8.6%±1.7%和3.8%±1.1%；P<0.05）。此外，白杨素（5.0、10.0和20.0 μmol/L）明显下调源自SKOV3的卵巢癌干细胞样细胞中CK2α、CD133和CD44的蛋白表达。联合CK2α siRNA转染，白杨素协同降低成球率（P<0.05）以及源自SKOV3的卵巢癌干细胞样细胞中CK2α、CD133和CD44的蛋白表达。然而，用pcDNA3.1-CK2α质粒转染减弱了白杨素对源自SKOV3的卵巢癌干细胞样细胞成球能力以及CK2α、CD133和CD44表达的抑制作用。
CK2α蛋白表达下调参与了白杨素对源自SKOV3的卵巢癌干细胞样细胞成球能力的抑制作用。
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