齐墩果酸对人卵巢癌SKOV3细胞增殖、侵袭、转移的影响及其作用机制研究

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<0.0...

Study on the Effects of Oleanolic Acid on the Proliferation,Invasion and Metastasis of Human Ovarian Cancer SKOV3 Cells and Its Mechanism

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 or P<0.01).In SKOV3 cells with over-expressed NF-κB p65,low-dose and high-dose of oleanolic acid could significantly down-regulat the protein expression of NF-κB p65,PRL-3,TNF-αand IL-6,while upregult the protein relative expression of E-cadherin(P<0.05 or P<0.01).CONCLUSIONS:Oleanolic acid can inhibit SKOV3 cells proliferation,invasion and metastasis by regulating NF-κB/PRL-3 signaling pathway.