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投稿时间：2020-02-26 网络发布日期：2020-10-20
投稿时间：2020-02-26 网络发布日期：2020-10-20
中文摘要: 目的 探讨口腔鳞癌细胞性别决定区域Y盒1(SOX1)、配对盒家族基因1(PAX1)表达及其对细胞增殖、侵袭的影响。
方法 体外培养人正常口腔组织细胞系、人口腔鳞癌细胞系HN4,采用RT-PCR及Western blot法检测细胞SOX1、PAX1 mRNA及蛋白表达水平;采用pCMV6-SOX 1、小干扰RNA(siRNA)进行细胞转染(不同转染获得空白1组、SOX1转染对照组、SOX1转染组和空白2组、PAX1转染对照组、PAX1转染组共6组细胞)分别上调SOX1的表达及下调PAX1的表达,采用噻唑蓝比色分析(MTT)法、划痕实验、Transwell法检测各组细胞增殖、迁移、侵袭行为,采用RT-PCR及Western blot检测Wnt通路信号分子-β-catenin及其下游效应分子(E-cadherin、MMP-2)等的表达。
结果 口腔鳞癌细胞SOX1、PAX1 mRNA和蛋白表达均显著低于正常口腔细胞（P均<0.05）。转染后，与空白1组及SOX1转染对照组比较，SOX1转染组细胞SOX1水平上调（P<0.05），细胞OD值和细胞侵袭数目降低（P均<0.05），细胞迁移减弱，在mRNA及蛋白水平上β-catenin、MMP-2表达降低（P均<0.05），E-cadherin表达升高（P均<0.05）；与空白2组及PAX1转染对照组比较，PAX1转染组口腔鳞癌细胞PAX1水平降低（P均<0.05），细胞OD值升高（P<0.05），细胞侵袭数目升高（P<0.05），迁移作用增强，在mRNA及蛋白水平上β-cateinin、MMP-2表达升高（P均<0.05），E-cadherin表达降低（P均<0.05）。
结论 SOX1、PAX1表达对口腔鳞癌细胞增殖、侵袭有一定影响,SOX1过表达可抑制细胞增殖、侵袭,而PAX1低表达可促进这一过程,两者对口腔鳞癌细胞生物学行为的影响可能是通过对Wnt信号通路及MMP-2的调控作用实现。
Abstract:Objective To explore the expressions of sex-determining region Y box 1 (SOX1) and paired boxed gene 1 (PAX1) in oral squamous cell carcinomas (OSCC) and their influences on cell proliferation and invasion.
Methods Human normal oral tissue cell line and human OSCC line HN4 were cultured in vitro.The mRNA and protein expression levels of SOX1 and PAX1 were detected by RT-PCR and Western blot.After transfecting cell by pCMV6-SOX1 and small interfering RNA (siRNA) (six groups of cells were obtained:blank 1 group,SOX1 transfection control group,SOX1 transfection group and blank 2 group,PAX1 transfection control group and PAX1 transfection group),SOX1 expression was up-regulated,and PAX1 expression was down-regulated respectively.Cell proliferation,migration and invasion were respectively detected by Methl thiazolyl tetrazolium (MTT),scratch test and Transwell method;RT-PCR and Western blot were used to detect the expressions of Wnt/β-catenin signaling pathway molecule and its downstream effector molecules (E-cadherin,MMP-2).
Results The mRNA and protein expressions of SOX1 and PAX1 in OSCC cells were significantly lower than those in normal oral cells (all P<0.05).After transfection,compared with the blank 1 group and SOX1 transfection control group,in SOX1 transfection group,the SOX1 level increased (P<0.05),the cell OD value and the number of cell invasion decreased (all P<0.05),cell migration effect was weakened,at mRNA and protein levels,the expression of β-catenin and MMP-2 decreased (all P<0.05),the expression of E-cadherin increased (all P<0.05);compared with the blank 2 group and the PAX1 transfection control group,in PAX1 transfection group,the level of PAX1 decreased (P<0.05),the cell OD value and the number of cell invasion increased (all P<0.05),cell migration effect was enhanced,at mRNA and protein levels,the expression of β-catenin and MMP-2 increased (all P<0.05),the expression of E-cadherin decreased (P<0.05).
Conclusions Over-expression of SOX1 can inhibit the proliferation and invasion of OSCC cells,while down-expression of PAX1 can promote this process.The effects of both SOX1 and PAX1 on biological behavior of OSCC cells may be achieved by regulating Wnt signaling pathway and MMP-2.
keywords: Sex-determining region Y box 1 Paired boxed gene 1 Oral squamous cell carcinoma Proliferation Invasion Wnt pathway
文章编号： 中图分类号： 文献标志码：A
基金项目:
| Author Name | Affiliation |
| LIU Chang,FU Yi-fu,LI Bing,WU Yi | Department of Clinical Laboratory,Nanjing Stomatological Hospital,Medical School of Nanjing University,Nanjing,Jiangsu 210008,China |
引用文本：
刘畅,符义富,李兵,吴仪.口腔鳞癌细胞SOX1、PAX1表达及其对细胞增殖、侵袭的影响[J].中国临床研究,2020,33(10):1308-1313.
刘畅,符义富,李兵,吴仪.口腔鳞癌细胞SOX1、PAX1表达及其对细胞增殖、侵袭的影响[J].中国临床研究,2020,33(10):1308-1313.