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-中国药房

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
阅读数：	239 次
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