胡桃醌对人卵巢癌SKOV3和IOSE80细胞增殖和侵袭的抑制作用及其机制研究
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摘要
目的:研究胡桃醌对人卵巢癌SKOV3和IOSE80细胞增殖和侵袭的抑制作用,并从信号通路深入探讨其作用机制。方法:体外培养人卵巢癌SKOV3细胞和IOSE80细胞,给予不同浓度的胡桃醌溶液干预,MTT比色法检测细胞增殖,划痕实验检测细胞迁移,Transwell检测细胞转移,流式细胞术检测细胞凋亡,Western Blot检测AKT、ERK、NF-κB信号通路蛋白表达,RT-PCR检测波形蛋白(vimentin)、基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9) mRNA表达。结果:不同浓度胡桃醌可抑制SKOV3细胞和IOSE80细胞增殖活力,降低细胞侵袭能力,加速细胞凋亡,且随浓度的增加对细胞增殖、侵袭和凋亡的作用越明显,差异均有统计学意义(P <0. 05)。不同浓度胡桃醌均可抑制SKOV3细胞和IOSE80细胞AKT、ERK、NF-κB信号通路蛋白的表达,且随浓度的增加信号通路蛋白表达减少,差异均有统计学意义(P <0. 05)。不同浓度胡桃醌均可减少SKOV3细胞和IOSE80细胞Vimentin、MMP-2、MMP-9 mRNA的表达,且随浓度的增加mRNA表达减少,差异均有统计学意义(P <0. 05)。结论:胡桃醌可抑制SKOV3和IOSE80细胞体外增殖、转移,促进凋亡,其机制与抑制AKT、ERK、NF-κB信号通路表达和下调Vimentin、MMP-2、MMP-9mRNA表达有关。
AIM:To study the effect of juglone on proliferation and invasion of human ovarian cancer SKOV3 and IOSE80 cells,and to explore its mechanism through signal pathway.METHODS:Cells were cultured in vitro and treated with different concentrations of juglone.The cell proliferation was detected by MTT colorimetry,the cell migration was detected by scratch test,the cell metastasis was detected by Transwell,and the apoptosis was detected by flow cytometry.The protein expression of AKT,ERK,NF-κB signaling pathway were detected by Western blot,and the expressions of Vimentin,MMP-2 and MMP-9 mRNA were detected by RTPCR.RESULTS:Different concentrations of juglone could inhibit the proliferation of SKOV3 and IOSE80 cells,reduce the invasive ability of cells,and accelerate the apoptosis,with increase of concentration the effect on cell proliferation,invasion and apoptosis were more obvious,the difference was statistically significant(P < 0.05).The expressions of AKT,ERK and NF-κB signaling pathway proteins in SKOV3 and IOSE80 cells were decreased by different concentrations of juglone,and the expression decreased with the increase of concentration,the difference was statistically significant(P < 0.05).The expressions of Vimentin,MMP-2 and MMP-9mRNA in SKOV3 and IOSE80 cells were decreased with the increase of concentration of juglone,and the difference was statistically significant(P <0.05).CONCLUSION:Juglone can inhibit the proliferation and metastasis of SKOV3 and IOSE80 cells in vitro and promote apoptosis.The mechanism maybe related to inhibit the expression of AKT,ERK,NF-κB signaling pathway,downregulate of the expression of Vimentin,MMP-2 and MMP-9mRNA.
AIM:To study the effect of juglone on proliferation and invasion of human ovarian cancer SKOV3 and IOSE80 cells,and to explore its mechanism through signal pathway.METHODS:Cells were cultured in vitro and treated with different concentrations of juglone.The cell proliferation was detected by MTT colorimetry,the cell migration was detected by scratch test,the cell metastasis was detected by Transwell,and the apoptosis was detected by flow cytometry.The protein expression of AKT,ERK,NF-κB signaling pathway were detected by Western blot,and the expressions of Vimentin,MMP-2 and MMP-9 mRNA were detected by RTPCR.RESULTS:Different concentrations of juglone could inhibit the proliferation of SKOV3 and IOSE80 cells,reduce the invasive ability of cells,and accelerate the apoptosis,with increase of concentration the effect on cell proliferation,invasion and apoptosis were more obvious,the difference was statistically significant(P < 0.05).The expressions of AKT,ERK and NF-κB signaling pathway proteins in SKOV3 and IOSE80 cells were decreased by different concentrations of juglone,and the expression decreased with the increase of concentration,the difference was statistically significant(P < 0.05).The expressions of Vimentin,MMP-2 and MMP-9mRNA in SKOV3 and IOSE80 cells were decreased with the increase of concentration of juglone,and the difference was statistically significant(P <0.05).CONCLUSION:Juglone can inhibit the proliferation and metastasis of SKOV3 and IOSE80 cells in vitro and promote apoptosis.The mechanism maybe related to inhibit the expression of AKT,ERK,NF-κB signaling pathway,downregulate of the expression of Vimentin,MMP-2 and MMP-9mRNA.
引文
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[17]赵行宇,杨森,周丽霞,等.胡桃醌对肝癌Hep G2细胞的增殖抑制及诱导凋亡作用[J].吉林大学学报(医学版),2013,39(2):255-258.
[18]Peng H,Wang Q,Qi X,et al.Orlistat induces apoptosis and protective autophagy in ovarian cancer cells:involvement of Akt-mTOR-mediated signaling pathway[J].Arch Gynecol Obstet,2018,298(3):597-605.
[19]王博,邵美丽,刘星,等.miR-148 a下调Livin促进卵巢癌OVCAR-3细胞的凋亡[J].疑难病杂志,2018,17(9):927-931.
[20]Hua F,Li CH,Chen XG.Daidzein exerts anticancer activity towards SKOV3 human ovarian cancer cells by inducing apoptosis and cell cycle arrest,and inhibiting the Raf/MEK/ERK cascade[J].Int J Mol Med,2018,41(6):3485-3492.
[21]Yang J,Li G.Pro-survival effects by NF-κB,Akt and ERK(1/2)and anti-apoptosis actions by Six1 disrupt apoptotic functions of TRAIL-Dr4/5 pathway in ovarian cancer[J].Biomed Pharmacother,2016,84(12):1078-1087.
[22]Han X,Zhou Y,You Y,et al.TET1 promotes cisplatin-resistance via demethylating the vimentin promoter in ovarian cancer[J].Cell Biol Int,2017,41(4):405-414.
[23]Sun X,Lin L,Chen Y,et al.Nitidine chloride inhibits ovarian cancer cell migration and invasion by suppressing MMP-2/9 production via the ERK signaling pathway[J].Mol Med Rep,2016,13(4):3161-3168.
[24]赵行宇,侯建成,金连海,等.胡桃醌对人宫颈癌He La细胞侵袭与迁移的影响[J].解剖学报,2017,48(2):160-164.
[2]Chen J,Jia Y,Jia ZH,et al.Silencing the expression of MTDH increases the radiation sensitivity of SKOV3ovarian cancer cells and reduces their proliferation and metastasis[J].Int J Oncol,2018,53(5):2180-2190.
[3]Ma Y,Xu Y.SPARCL1 suppresses the proliferation and migration of human ovarian cancer cells via the MEK/ERK signaling[J].Exp Ther Med,2018,16(4):3195-3201.
[4]Li X,Bao C,Ma Z,et al.Perfluorooctanoic acid stimulates ovarian cancer cell migration,invasion via ERK/NF-κB/MMP-2/-9 pathway[J].Toxicol Lett,2018,294(9):44-50.
[5]Mortensen JH,Godskesen LE,Jensen MD,et al.Fragments of citrullinated and MMP-degraded vimentin and MMP-degraded type III collagen are novel serological biomarkers to differentiate crohn's disease from ulcerative colitis[J].J Crohns Colitis,2015,9(10):863-872.
[6]王添敏,俞文婕,邸学,等.加入胡桃醌与敲除胡桃醌的胡桃楸水提物的抗肿瘤活性[J].西北药学杂志,2017,32(4):480-485.
[7]Lu Z,Chen H,Zheng XM.Experimental study on the apoptosis of cervical cancer Hela cells induced by juglone through c-Jun N-terminal kinase/c-Jun pathway[J].Asian Pac J Trop Med,2017,10(6):572-575.
[8]方芳,赵良中,李强,等.胡桃醌对卵巢癌SKOV3细胞氧化应激和凋亡的影响[J].中国病理生理杂志,2016,32(1):156-159.
[9]Moore KN.Administration of the Tablet Formulation of Olaparib in Patients with Ovarian Cancer:Practical Guidance and Expectations[J].Oncologist,2018,23(6):697-703.
[10]Di Lorenzo G,Ricci G,Severini GM,et al.Imaging and therapy of ovarian cancer:clinical application of nanoparticles and future perspectives[J].Theranostics,2018,8(16):4279-4294.
[11]李冀,张睿苏,高彦宇,等.半枝莲含药血清对人卵巢癌SKOV3细胞增殖的影响[J].中医药信息,2016,33(3):1-4.
[12]Jin X,Zhang Y,Zhang Z,et al.Juglone loaded poloxamer 188/phospholipid mixed micelles evaluated in vitro and in vivo in breast cancer[J].Int J Pharm,2016,515(1/2):359-366.
[13]Peng X,Nie Y,Wu J,et al.Juglone prevents metabolic endotoxemia-induced hepatitis and neuroinflammation via suppressing TLR4/NF-κB signaling pathway in high-fat diet rats[J].Biochem Biophys Res Commun,2015,462(3):245-250.
[14]李岩,赵剑璞,王燕妮,等.胡桃醌含药血清体外抗肿瘤实验研究[J].北京理工大学学报,2013,33(5):545-550.
[15]董文婷,霍金海,张海燕,等.干、鲜北青龙衣中胡桃醌的含量及抗肿瘤活性比较[J].吉林中医药,2016,36(4):401-404.
[16]方芳,郝峰,李强,等.胡桃醌抑制卵巢癌SKOV3细胞黏附和侵袭作用[J].中国公共卫生,2015,31(9):1159-1161.
[17]赵行宇,杨森,周丽霞,等.胡桃醌对肝癌Hep G2细胞的增殖抑制及诱导凋亡作用[J].吉林大学学报(医学版),2013,39(2):255-258.
[18]Peng H,Wang Q,Qi X,et al.Orlistat induces apoptosis and protective autophagy in ovarian cancer cells:involvement of Akt-mTOR-mediated signaling pathway[J].Arch Gynecol Obstet,2018,298(3):597-605.
[19]王博,邵美丽,刘星,等.miR-148 a下调Livin促进卵巢癌OVCAR-3细胞的凋亡[J].疑难病杂志,2018,17(9):927-931.
[20]Hua F,Li CH,Chen XG.Daidzein exerts anticancer activity towards SKOV3 human ovarian cancer cells by inducing apoptosis and cell cycle arrest,and inhibiting the Raf/MEK/ERK cascade[J].Int J Mol Med,2018,41(6):3485-3492.
[21]Yang J,Li G.Pro-survival effects by NF-κB,Akt and ERK(1/2)and anti-apoptosis actions by Six1 disrupt apoptotic functions of TRAIL-Dr4/5 pathway in ovarian cancer[J].Biomed Pharmacother,2016,84(12):1078-1087.
[22]Han X,Zhou Y,You Y,et al.TET1 promotes cisplatin-resistance via demethylating the vimentin promoter in ovarian cancer[J].Cell Biol Int,2017,41(4):405-414.
[23]Sun X,Lin L,Chen Y,et al.Nitidine chloride inhibits ovarian cancer cell migration and invasion by suppressing MMP-2/9 production via the ERK signaling pathway[J].Mol Med Rep,2016,13(4):3161-3168.
[24]赵行宇,侯建成,金连海,等.胡桃醌对人宫颈癌He La细胞侵袭与迁移的影响[J].解剖学报,2017,48(2):160-164.