尿路上皮癌胚抗原1对口腔癌细胞侵袭迁移及增殖影响的实验研究  

The influence of urothelial carcinoembryonic antigen 1 on invasion and migration of oral carcinoma cell lines

作　　者：杨勇涛[1] 杨宏宇[1] 王宇帆[1] 沈时岳[1] 李明华[2] 秦嘉若[1] 

机构地区：[1]北京大学深圳医院口腔颌面外科,广东518036 [2]北京大学深圳医院中心实验室,广东518036

出　　处：《中华口腔医学杂志》2016年第1期36-41,共6页Chinese Journal of Stomatology

基　　金：国家自然科学基金（81572654）;广东省自然科学基金（S2012010010382）

摘　　要：目的 探讨长链非编码RNA尿路上皮癌胚抗原1（urothelial carcinoembryonic antigen 1,UCA1）对口腔癌细胞系SCC15和CAL27侵袭、迁移及增殖能力的影响.方法 瞬时转染UCA1-siRNA沉默SCC15和CAL27细胞中长链非编码RNA UCA1的表达,以阴性干扰RNA为对照组,在实验的5、24、48、72、96 h为观测点.利用实时荧光定量核酸扩增反应（quantitative real time,qRT）-PCR方法检测UCA1干扰效果;通过蛋白质印迹法检测SCC15和CAL27中UCA1实验组及对照组基质金属蛋白酶（matrix metalloproteinase 9,MMP-9）蛋白表达水平;通过划痕实验及Transwell实验检测其侵袭及迁移能力;通过CCK-8实验检测细胞增殖能力.结果 SCC15和CAL27细胞的UCA1有效沉默,SCC15和CAL27两株细胞系UCA1干扰效率分别为86.45％（P＜0.001）和78.24％ （P＜0.001）;沉默后细胞的迁移侵袭及增殖能力均减弱,CAL27对照组与实验组的迁移、侵袭穿膜细胞分别为（719.20±92.36）和（208.00±25.58）个（P=0.000 7）;（363.40±45.96）和（164.80±24.68）个（P=0.005 2）;SCC15对照组与实验组的迁移、侵袭穿膜细胞分别为（437.20±54.75）和（145.80±23.31）个（P=0.001 1）、（249.80±38.41）和（63.80±11.11）个（P=0.001 6）;通过检测并计算出实验组相对于对照组各个时间点的相对增殖率,即实验组中24、48、72和96 h与对照组中同一时间点比较,SCC 15：R24 h=0.870、R48 h=0.863、R72h=0.643、R96h=0.732;CAL27：R24h=0.913、R48 h=0.829、R72 h=0.756、R96 h=0.705） （P＜0.05）;MMP-9蛋白表达量下降.结论 UCA1能通过调节MMP-9蛋白的表达增强其侵袭迁移能力,并能促进其增殖能力,从而在口腔癌侵袭进展中发挥作用.Objective To investigate the effects of long chain non-coding RNA urothelial carcinoembryonic antigen 1（UCA1） on invasion, migration and proliferation abilities in oral squamous cell carcinoma cell lines SCC15 and CAL27.Methods Small interfering RNA of UCA1（UCA1-siRNA） was transfected into SCC15 and CAL27 cell lines by LipofectamineTM 3000 to silence UCA1, and transfected negtive control si-RNA served as a control.The effect of UCA 1-siRNA was detected by quantitative real time-polymerase chain reaction（qRT-PCR） to confirm the successful inhibition of UCA1 by siRNA.The matrix metalloproteinase 9（MMP-9） protein level was detected by Western blotting analysis.The effect of siRNA on cell proliferation and invasion was assessed by transwell migration assay and wound healing assay.Cell counting kit-8（CCK-8） assay was carried out to estimate the proliferation of two cell lines with different expression levels of UCA1.Results Expressions of UCA1 of SCC15 and CAL27 were successfully suppressed after transfected with siRNA which verified by qRT-PCR, and the efficiency of downregulation of SCC15 and CAL27 was 86.45%（P〈0.001）and 78.24%（P〈0.001）, respectively.The migration, invasion and proliferation of SCC15 and CAL27 cell lines after transfected with siRNA were obviously restrained.The number of migration and invasion of CAL27 cells were 719.20±92.36 versus 208.00±25.58 （P=0.000 7） and 363.40±45.96 versus 164.80±24.68（P=0.005 2）, respectively, the number of migration and invasion of SCC15 cells were 437.20±54.75 vs 145.80±23.31（P=0.001 1） and 249.80±38.41 vs 63.80± 11.11 （P=0.001 6）, respectively （UCA1-si compare to negtive control）, the relative proliferation rates of SCC 15 and CAL27 were SCC15：R24 h=0.870, R48 h=0.863, R72 h=0.64, R96 h=0.732;CAL27： R24 h=0.913, R48 h=0.829, R72 h=0.756, R96 h=0.705（P〈0.05）, and MMP-9 expression level was decreased by UCA1-siRNA compared with negative control.Conclusions UCA1 could enhance the ability of invasion and

关 键 词：口腔鳞状细胞癌 口腔肿瘤 尿路上皮癌胚抗原1 UROTHELIAL carcinoembryonic ANTIGEN 1 

分 类 号：R739.85[医药卫生—肿瘤]