口腔癌前病变细胞中尼古丁诱导α3nAChR、α7nAChR表达及与Prx1相互作用研究  

The interaction betweenα3nAChR andα7nAChR and Prx1 in nicotine-induced oral precancerous lesion cells

作　　者：齐墨词 沈亚俊 汤晓飞[1] 张敏[1] QI Mo-ci;SHEN Ya-jun;TANG Xiao-fei;ZHANG Min(Beijing Institute of Dental Research,Capital Medical University School of Stomatology,Beijing 100050,China)

机构地区：[1]首都医科大学口腔医学院口腔医学研究所,北京100050

出　　处：《北京口腔医学》2022年第4期229-233,共5页Beijing Journal of Stomatology

基　　金：国家自然科学基金(81470752,82071114);北京市教委科技计划项目(KM201910025009)。

摘　　要：目的研究口腔癌前病变细胞中尼古丁诱导α3n AChR和α7nAChR表达及与Prx1的相互作用,探讨尼古丁在口腔癌前病变中的作用机制。方法利用Discovery Studio 2016软件预测α3nAChR和α7n AChR与Prx1是否存在直接结合,在体外培养的口腔癌前病变DOK细胞中进行验证。细胞分为尼古丁组(1μmol/L尼古丁处理7 d)和对照组(无任何处理),利用Co-IP和Western Blot检测细胞中Prx1与α3nAChR或α7nAChR的结合。采用α-BTX阻断α7nAChR受体,Western Blot检测尼古丁诱导Prx1结合蛋白GTPBP4和DIRAS2的表达。结果尼古丁可诱导DOK细胞α3nAChR、α7nAChR、Prx1及其结合蛋白GTPBP4和DIRAS2表达增高。软件预测α3nAChR和α7nAChR均可能通过氢键和疏水键与Prx1直接结合,但是在DOK细胞中未检测到Prx1与α3nAChR或α7nAChR的结合。α-BTX特异性阻断α7n AChR对DOK细胞中GTPBP4和DIRAS2的蛋白水平无明显影响(P>0.05)。结论尼古丁可诱导口腔癌前病变细胞α3nAChR、α7n AChR、Prx1及其结合蛋白GTPBP4和DIRAS2表达增高;α7nAChR对Prx1的调控并非通过两者直接结合发挥作用,且亦非通过GTPBP4、DIRAS2介导完成。Objective To investigate the interaction between nicotinic acetylcholine receptors(nAChRs)α3nAChR andα7n ACh R and peroxiredoxin 1(Prx1)in nicotine-induced oral precancerous lesion cells.Methods The Discovery Studio 2016 was used to predict the direct interaction betweenα3nAChR andα7nAChR and Prx1.Human oral precancerous DOK cells were cultured and divided into nicotine group and control group.The nicotine group was treated with 1μmol/L nicotine for 7 days while the control group was untreated.Western Blot and Co-IP were performed to verify whether Prx1 interacted withα7nAChR andα3nAChR.In nicotine group,DOK cells were treated withα-BTX,a specific inhibitor ofα7nAChR.Prx1 binding proteins GTPBP4 and DIRAS2 expression were detected by Western Blot.Results Discovery Studio 2016predicted that there may be a direct binding ofα3nAChR andα7n AChR to Prx1,but no interaction was detected between Prx1 andα3nAChR andα7nAChR in DOK cells.By usingα-BTX to specifically blockα7nAChR,compared with nicotine group,the expression of GTPBP4 and DIRAS2 in nicotine+α-BTX group did not change significantly(P>0.05).Conclusions Nicotine can induce increased expression ofα3n ACh R,α7n ACh R,Prx1 and its binding proteins GTPBP4 and DIRAS2 in oral precancerous cells.The regulation ofα7n AChR on Prx1 is mediated by neither directly binding nor GTPBP4and DIRAS2 expression.

关 键 词：过氧化物还原酶1 乙酰胆碱受体 口腔癌前病变 尼古丁 

分 类 号：R739.8[医药卫生—肿瘤]