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
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篇名: 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
TITLE: Study on the Effects of Oleanolic Acid on the Proliferation ,Invasion and Metastasis of Human Ovarian Cancer SKOV3 Cells and Its Mechanism
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、48h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24h对SKOV3细胞迁移及侵袭能力的影响;采用WesternBlotting法检测低、高剂量齐墩果酸对SKOV3细胞中核因子κBp65(NF-κBp65)、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-κBp65质粒转染SKOV3细胞,采用Westernblotting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-κBp65、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-κBp65、PRL-3、TNF-α、IL-6蛋白及其mRNA的相对表达量较对照组显著升高,E-cadherin蛋白及其mRNA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-κBp65、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mRNA的相对表达量均显著升高(P<0.05或P<0.01)。在NF-κBp65过表达SKOV3细胞中,低、高剂量齐墩果酸同样能够显著下调NF-κBp65、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。
ABSTRACT: OBJECTIVE:To investigate the effects and m echanism of oleanolic acid on inhibiting the proliferation ,invasion and metastasis of human ovarian cancer SKOV 3 cells. METHODS :CCK-8 assay was used to detect the effects of different concentrations of oleanolic acid (10,20,40,60,80,100 μmol/L)on the proliferation of ovarian cancer SKOV 3 cells at 12,24, 36 and 48 h. The effects of low-dose and high-dose of oleanolic acid (20,40 μmol/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 SKOV 3 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 SKOV 3 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-α and IL-6 in SKOV 3 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 mRNA relative expression of NF-κB p65,PRL-3,TNF-α and 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-κ B p65,PRL-3,TNF-α and 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 c ould significantly down-regulat the proteinexpression of NF-κ B p65,PRL-3,TNF-α and IL-6,while upregult the protein relative expression of E-cadherin (P<0.05 E-mail:122821905@qq.com or P<0.01). CONCLUSIONS : Oleanolic acid can inhibit SKOV3 cells proliferation,invasion and metastasis by regulating NF-κB/PRL-3 signaling pathway.
期刊: 2020年第31卷第10期
作者: 杜贵强,张永莉,胡孝辉
AUTHORS: DU Guiqiang ,ZHANG Yongli ,HU Xiaohui
关键字: 齐墩果酸;卵巢癌;SKOV3细胞;增殖;侵袭;转移;核因子κB;肝再生磷酸酶3
KEYWORDS: Oleanolic acid ;Ovarian cancer ;SKOV3 cell;Proliferation;Invasion;Metastasis;NF-κB;PRL-3
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