摘要
目的探讨山姜素对人卵巢癌OVCAR-8细胞的增殖、迁移的抑制作用及其可能机制。方法以DMSO为溶剂对照,使用不同浓度山姜素(25、50、100、200和400μmol/L)处理OVCAR-8不同时间(24、48和72h)后,采用CCK-8试剂盒检测细胞增殖活力;采用3D肿瘤微球形成实验检测50μmol/L山姜素处理7d后肿瘤微球形成能力;使用划痕实验检测100μmol/L山姜素处理24 h后对迁移能力;采用Western blot法检测不同浓度山姜素(50、100和200μmol/L,以DMSO为对照)作用48 h后OVCAR-8细胞STAT3信号通路及凋亡相关蛋白表达变化。结果 25~400μmol/L山姜素对OVCAR-8细胞增殖均有抑制作用,且呈剂量-时间依赖效应关系(均P<0.05);50μmol/L山姜素处理组肿瘤微球直径较DMSO组明显减小(P<0.05),而100μmol/L山姜素组划痕宽度明显宽于DMSO对照组(P<0.05);此外,50、100和200μmol/L山姜素处理组Bax、cleaved-caspase-3和-9的表达明显增加,Bcl-2、p-STAT3、c-myc、survivin蛋白表达明显减少,均呈一定的剂量依赖效应关系(均P<0.05)。而STAT3蛋白水平无明显变化(P>0.05)。结论山姜素对卵巢癌OVCAR-8细胞增殖和迁移具有抑制作用,且可能与STAT3信号通路抑制以及线粒体凋亡通路激活相关。
Objective To explore the effect of alpinetin on human ovarian cancer cell line OVCAR-8 and its mecha nism. Methods OVCAR-8 cells were treated with different concentrations of alpinetin(25, 50, 100, 200 and 400μmol/L,DMSO as control) for different hours(24, 48 and 72h), then cell viabilities were detected by CCK-8 assays. 3D spheroid assay was used to measure the spheroid formation capacity of OVCAR-8 cell after treated with 50 μmol/L alpinetin(DMSO as control) for 7 days. Besides, wound healing assay was used to measure the migration capacity of OVCAR-8 cell after treated with 100 μmol/L alpinetin(DMSO as control) for 24h. In addition, Western blot was used to detect the protein levels of STAT3 signaling related and apoptosis related proteins in OVCAR-8 cells after alpinetin treatments(50, 100 and 200μmol/L, DMSO as control) for 48h. Results 25-400μmol/L alpinetin had remarkable inhibiting effects on OVCAR-8 cells and in a dose-and time-dependent manner(all P〈0.05). After 7 days′ treatment, the diameter of spheroid in 50 μmol/L alpinetin group was significantly shorter than DMSO control group(P〈0.05). Besides, the width of wound in 100 μmol/L alpinetin group was significantly longer than DMSO control group after treatment for 24h(P〈0.05). In addition, the protein levels of Bcl-2, p-STAT3, c-myc and survivin were decreased in 50,100 and 200 μmol/L group of OVCAR-8 cells than DMSO control group after 48 exposure. On the contrary, the protein level of Bax, cleaved-caspase-3 and-9 was increased(all P〈0.05), and there were no significantly changes in STAT3 levels(P〈0.05). Conclusion Alpinetin can inhibit the proliferation and migration of OVCAR-8 cells,its mechanism may be related to the inhibition of STAT3 signaling pathway and the activation of mitochondrial apoptosis signaling.
作者
曾静
王璟
殷金凤
宋琦
ZENG Jing ,WANG Jing, YIN Jinfeng ,SONG Qi(Department of Gynecology and Obstetrics, Chengdu Second People's Hospital, Sichuan Province,Chengdu 610017, Chin)
出处
《中国医药导报》
CAS
2018年第10期13-17,共5页
China Medical Herald
关键词
山姜素
卵巢癌
增殖
迁移
微球
凋亡
Alpinetin
Ovarian cancer
Proliferation
Migration
Spheroid
Apoptosis