姜玲, 曹晓正, 李程. The effects of casticin on phosphorylation of FoxO3a in stem-like cells derived from ovarian cancer SKOV3 cell line[J]. China Oncology, 2015, 25(5): 360-364.
姜玲, 曹晓正, 李程. The effects of casticin on phosphorylation of FoxO3a in stem-like cells derived from ovarian cancer SKOV3 cell line[J]. China Oncology, 2015, 25(5): 360-364. DOI: 10.3969/j.issn.1007-3969.2015.05.007.
背景与目的:肿瘤干细胞的存在是卵巢癌高复发率和高死亡率的原因。本文研究紫花牡荆素(casticin,CAS)对SKOV3细胞系卵巢癌干细胞样细胞(ovarian cancer stem cell like cells,OCSLCs)自我更新能力的影响及其机制。方法:体外培养SKOV3细胞系细胞,干细胞培养基悬浮培养SKOV3得到和扩增卵巢癌球形成细胞,即OCSLCs。蛋白[质]印迹法(Western blot)分析FoxO3a磷酸化水平。FoxO3a特异性siRNA转染抑制FoxO3a蛋白表达,并测定肿瘤球形成率。结果:与亲本细胞相比,OCSLCs高表达磷酸化FoxO3a蛋白,具有更高的肿瘤球形成率[(3.1±0.3)% vs (34.8±6.8)%,P<0.05]。CAS显著抑制OCSLCs肿瘤球形成能力,并降低FoxO3a磷酸化水平。FoxO3a特异性siRNA转染抑制FoxO3a蛋白表达,拮抗CAS抑制OCSLCs肿瘤球形成作用。结论:降低FoxO3a磷酸化水平参与CAS抑制SKOV3细胞系OCSLCs肿瘤球形成作用。
Abstract
Background and purpose: Ovarian cancer is associated with a high recurrence and mortality due to the existence of cancer stem cells. The purpose of this study was to investigate the effect of casticin (CAS) on the capability of self-renewal in ovarian cancer stem cell like cells (OCSLCs) derived from SKOV3 cell line. Methods: SKOV3 cell line cells were cultured in vitro
and OCSLCs were obtained and amplified through suspended culture with conditioned medium of the stem cells. The phosphorylation level of FoxO3a was analyzed using Western blot. The protein expression of FoxO3a was inhibited by FoxO3a specific siRNA transfection
and then the ratio of sphereformation was detected. Results: Compared with parental cells
OCSLCs over-expressed phosphorylated FoxO3a (pFoxO3a) and had elevated ratio of sphere-formation [(3.1±0.3)% vs (34.8±6.8)%
P0.05]. CAS significantly inhibited the capability of sphere-formation in OCSLCs and down-regulated the expression level of pFoxO3a. And the transfection of FoxO3a specific siRNA suppressed the protein expression of FoxO3a and attenuated the inhibitory effect of CAS on the sphere-formation of OCSLCs. Conclusion: Reduced expression level of pFoxO3a is involved in the effect that CAS inhibits sphere-formation of OCSLCs derived from SKOV3 cell line.