摘要
目的:探讨齐墩果酸抑制人卵巢癌SKOV3细胞增殖、侵袭、转移的作用及其机制。方法:采用CCK-8法检测不同浓度齐墩果酸(10、20、40、60、80、100μmol/L)作用12、24、36、48 h对卵巢癌SKOV3细胞增殖能力的影响;采用Transwell实验观察低、高剂量齐墩果酸(20、40μmol/L)作用24 h对SKOV3细胞迁移及侵袭能力的影响;采用Western Blotting法检测低、高剂量齐墩果酸对SKOV3细胞中核因子κB p65(NF-κB p65)、肝再生磷酸酶3(PRL-3)、肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)、上皮钙黏着蛋白E(E-cadherin)等蛋白表达的影响;运用脂多糖(LPS)诱导和NF-κB p65质粒转染SKOV3细胞,采用Western blotting法和实时荧光定量PCR考察低、高剂量齐墩果酸对NF-κB/PRL-3通路相关蛋白及其mRNA表达的影响。结果:随着齐墩果酸给药浓度的增加和作用时间的延长,SKOV3细胞的增殖能力有随之降低的趋势,各剂量组的细胞存活率均显著低于对照组(P<0.05或P<0.01)。低、高剂量的齐墩果酸组的迁移和侵袭细胞数均显著减少(P<0.05或P<0.01),且其NF-κB p65、PRL-3、TNF-α、IL-6蛋白的相对表达量均显著降低,E-cadherin蛋白的相对表达量均显著升高(P<0.05或P<0.01)。经LPS刺激后,LPS模型组细胞中NF-κB p65、PRL-3、TNF-α、IL-6蛋白及其m RNA的相对表达量较对照组显著升高,E-cadherin蛋白及其m RNA的相对表达量均显著降低(P<0.05或P<0.01);低、高剂量齐墩果酸组细胞中NF-κB p65、PRL-3、TNF-α、IL-6蛋白及其mRNA的相对表达量均显著降低,E-cadherin蛋白及其m RNA的相对表达量均显著升高(P<0.05或P<0.01)。在NF-κB p65过表达SKOV3细胞中,低、高剂量齐墩果酸同样能够显著下调NF-κB p65、PRL-3、TNF-α、IL-6蛋白的表达,显著上调E-cadherin蛋白的表达(P<0.05或P<0.01)。结论:齐墩果酸能够通过调控NF-κB/PRL-3信号通路来抑制人卵巢癌细胞SKOV3的增殖、侵袭、转移。
OBJECTIVE:To investigate the effects and mechanism of oleanolic acid on inhibiting the proliferation,invasion and metastasis of human ovarian cancer SKOV3 cells.METHODS:CCK-8 assay was used to detect the effects of different concentrations of oleanolic acid(10,20,40,60,80,100μmol/L)on the proliferation of ovarian cancer SKOV3 cells at 12,24,36 and 48 h.The effects of low-dose and high-dose of oleanolic acid(20,40μmol/L)on the metastasis and invasion ability of SKOV3 cells for 24 h were observed in Transwell assay.Western blotting assay was used to detect the effects of low-dose and high-dose of oleanolic acid on the protein expression of NF-κB p65,PRL-3,TNF-α,IL-6 and E-cadherin in SKOV3 cells.Through LPS induction and NF-κB p65 plasmid transfection,Western blotting and RT-qPCR assay were used to investigate the effects of low-dose and high-dose oleanolic acid on the expression of NF-κB/PRL-3 pathway related proteins and their mRNA.RESULTS:With the increase of the concentration and action time of oleanolic acid,the proliferation capacity of ovarian cancer SKOV3 cells was decreased,the surval rates of administration groups were significantly lower than that of the control group(P<0.05 or P<0.01).Low-dose and high-dose of oleanolic acid could significantly reduce the number of migrating and invading cells(P<0.05 or P<0.01).The protein relative expression of NF-κB p65,PRL-3,TNF-αand IL-6 in SKOV3 cells were significantly decreased,while the protein relative expression of E-cadherin was significantly increased(P<0.05 or P<0.01).After LPS induction,protein and m RNA relative expression of NF-κB p65,PRL-3,TNF-αand IL-6 were increased significantly in LPS model group,while protein and mRNA relative expression of E-cadherin were significantly decreased(P<0.05 or P<0.01).The protein and mRNA relative expression of NF-κB p65,PRL-3,TNF-αand IL-6 were significantly decreased,and protein and mRNA relative expression of E-cadherin were significantly increased in low-dose and high-dose of oleanolic acid group(P<0.05
作者
杜贵强
张永莉
胡孝辉
DU Guiqiang;ZHANG Yongli;HU Xiaohui(Dept.of Gynecology,the First Affiliated Maternal and Infant Hospital of Tongji University,Shanghai 200126,China)
出处
《中国药房》
CAS
北大核心
2020年第10期1190-1197,共8页
China Pharmacy
基金
国家自然科学基金资助项目(No.81871126)。
