β-谷甾醇诱导口腔鳞状细胞癌SCC9细胞凋亡及机制研究
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摘要
目的:研究_β-谷甾醇对口腔鳞状细胞癌(oral squamous cell carcinoma,OSCC)细胞SCC9的作用及分子机制,以期为_β-谷甾醇应用于临床治疗OSCC等肿瘤提供理论支持。方法:采用不同浓度的_β-谷甾醇体外作用于培养中的SCC9细胞,观察SCC9的凋亡发生情况,同时检测SCC9胞浆内活性氧(reactive oxygen species,ROS)水平改变及烟酰胺腺嘌呤二核苷酸磷酸(nicotinamide adenine dinucleotide phosphate,NADPH)氧化酶gp91~(phox)基因及蛋白表达水平;通过小干扰RNA(small interference RNA,siRNA)技术抑制SCC9细胞中gp91~(phox)的表达对其功能进行确认。结果:结果显示,相较于对照组细胞,β-谷甾醇显著诱导其凋亡,且呈浓度依赖性(P<0.01);β-谷甾醇显著提高SCC9胞浆内ROS水平,促进gp91~(phox)的表达(P<0.01);抑制gp91~(phox)表达可阻断_β-谷甾醇对SCC9的促凋亡作用(P<0.01)。结论:β-谷甾醇可有效诱导SCC9细胞凋亡,且NADPH氧化酶gp91~(phox)在其中起关键调控作用,可作为临床治疗的潜在靶点之一。
Objective:To investigate the effects and related mechanism ofβ-sitosterol on oral squamous cell carcinoma cells SSC9.Methods:SCC9cells were treated by various dose ofβ-sitosterol,and the cell apoptosis was monitored.The intracellular ROS were analyzed and the mRNA and protein level of NADPH oxidase gp91~(phox )were detected.Knock down of gp91~(phox )in SCC9was performed to confirm the effects ofβ-sitosterol on SCC9cells.Results:Compared with the control group,β-sitosterol induced cell apoptosis in a dose dependent manner(P<0.01).β-sitosterol elevated the intracellular ROS level and mRNA and protein expression of gp91~(phox )in SCC9cells.Inhibition of gp91~(phox )blocked the effect ofβ-sitosterol on SCC9cells.Conclusion:NADPH oxidase gp91~(phox )had an important role in mediating theβ-sitosterol-induced apoptotic process in SCC9cells,which might be a target of OSCC treatment.
Objective:To investigate the effects and related mechanism ofβ-sitosterol on oral squamous cell carcinoma cells SSC9.Methods:SCC9cells were treated by various dose ofβ-sitosterol,and the cell apoptosis was monitored.The intracellular ROS were analyzed and the mRNA and protein level of NADPH oxidase gp91~(phox )were detected.Knock down of gp91~(phox )in SCC9was performed to confirm the effects ofβ-sitosterol on SCC9cells.Results:Compared with the control group,β-sitosterol induced cell apoptosis in a dose dependent manner(P<0.01).β-sitosterol elevated the intracellular ROS level and mRNA and protein expression of gp91~(phox )in SCC9cells.Inhibition of gp91~(phox )blocked the effect ofβ-sitosterol on SCC9cells.Conclusion:NADPH oxidase gp91~(phox )had an important role in mediating theβ-sitosterol-induced apoptotic process in SCC9cells,which might be a target of OSCC treatment.
引文
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[10]夏冬景,黄莹莹,李海朋,等.磷酸二酯酶4D在口腔鳞状细胞癌发生发展过程中的作用研究[J].口腔医学研究,2017,33(6):663-667.
[2]Bin Sayeed MS,Ameen SS.Beta-Sitosterol:apromising but orphan nutraceutical to fight against cancer[J].Nutr Cancer,2015,67(8):1214-1220.
[3]Mahaddalkar T,Suri C,Naik PK,et al.Biochemical characterization and molecular dynamic simulation ofβ-sitosterol as a tubulin-binding anticancer agent[J].Eur J Pharmacol,2015,760(2):154-162.
[4]Wu AH,Ruan W,Todd J,et al.Biological variation ofβ-sitosterol,campesterol,and lathosterol as cholesterol absorption and synthesis biomarkers[J].Clin Chim Acta,2014,430(9):43-47.
[5]Bodner L,Manor E,Friger MD,et al.Oral squamous cell carcinoma in patients twenty years of age or younger--review and analysis of 186reported cases[J].Oral Oncol,2014,50(2):84-89.
[6]Malik UU,Zarina S,Pennington SR.Oral squamous cell carcinoma:Key clinical questions,biomarker discovery,and the role of proteomics[J].Arch Oral Biol,2016,63:53-65.
[7]Roy K,Wu Y,Meitzler JL,et al.NADPH oxidases and cancer[J].Clin Sci,2015,128(5):863-875.
[8]Lee Y,Kwak HB,Hord J,et al.Exercise training attenuates age-dependent elevation of angiotensinⅡtype 1receptor and Nox2signaling in the rat heart[J].Exp Gerontol,2015,70:163-173.
[9]Ishaq M,Evans MD,Ostrikov KK.Atmospheric pressure gas plasma-induced colorectal cancer cell death is mediated by Nox2-ASK1apoptosis pathways and oxidative stress is mitigated by Srx-Nrf2anti-oxidant system[J].Biochim Biophys Acta,2014,1843(12):2827-2837.
[10]夏冬景,黄莹莹,李海朋,等.磷酸二酯酶4D在口腔鳞状细胞癌发生发展过程中的作用研究[J].口腔医学研究,2017,33(6):663-667.