miR-34a通过下调SATB2抑制口腔鳞癌增殖、迁移和侵袭的初步研究  

MiR-34a down-regulates SATB2 expression to inhibit the proliferation,migration and invasion of oral squamous cell carcinoma

作　　者：葛昕[1,2] 詹甜甜 高洁[1,2] 张勃昕[1,2] 王晨星 李怀奇[1,2] 叶金海[1,2] GE Xin;ZHAN Tiantian;GAO Jie;ZHANG Boxin;WANG Chenxing;LI Huaiqi;YE Jinhai(Jiangsu Key Laboratory of Oral Diseases,Nanjing Medical University,Nanjing 210029,China;Department of Oral and Maxillofacial Surgery,Affiliated Hospital of Stomatology,Nanjing Medical University,Nanjing 210029,China)

机构地区：[1]南京医科大学口腔疾病研究江苏省重点实验室,江苏南京210029 [2]南京医科大学附属口腔医院口腔颌面外科,江苏南京210029

出　　处：《口腔生物医学》2018年第1期1-6,16,共7页Oral Biomedicine

基　　金：国家自然科学基金资助项目(30801301;81371123);江苏省"青蓝工程"项目;"科教强卫工程"医学重点人才项目;江苏高校优势学科建设工程资助项目(2014-37)

摘　　要：目的:检测mi R-34a在口腔鳞癌组织中的表达,探讨其是否可以通过调节SATB2的表达影响口腔鳞癌的增殖、迁移和侵袭。方法:采用实时定量RT-PCR的方法检验口腔鳞癌和正常组织、细胞中mi R-34a基因的表达水平;在过表达SATB2的HN6细胞中转染mi R-34a mimic质粒后,采用CCK-8实验检测细胞增殖能力的变化,流式细胞术检验细胞周期,细胞划痕实验和Transwell实验检测细胞迁移能力的变化,细胞侵袭实验检测mi R-34a对细胞侵袭能力的影响;同时提取细胞内总蛋白,采用Western blot的方法检测与侵袭相关的基质金属蛋白酶(MMP)-2、MMP-9以及SATB2的表达;通过裸鼠荷瘤模型检验mi R-34a对SATB2表达及移植瘤生长的影响。结果:mi R-34a在口腔鳞癌组织和口腔鳞癌细胞系HN6中呈低表达,而在癌旁组织和人口腔角质形成细胞中呈高表达,差异具有统计学意义(P<0.05);在过表达SATB2的HN6细胞中转染mi R-34a mimic质粒后,细胞的增殖,迁移和侵袭能力明显减弱;在裸鼠皮下荷瘤实验中,转染了mi R-34a mimic质粒的过表达SATB2的HN6细胞所成的瘤体较对照组明显减小(P<0.05)。结论:mi R-34a可以通过抑制SATB2的表达来抑制口腔鳞癌的增殖、迁移和侵袭。Objective:To investigate expression of miR-34a in oral squamous cell carcinoma(OSCC)tissue samples,and then to study whether it can inhibit the proliferation,migration and invasion of oral squamous cell carcinoma by down-regulating the expression of SATB2.Methods:The expression of miR-34a in OSCC,HN6 cell line was tested by qRT-PCR.We used lentivirus to overexpress SATB2 and constructed miR-34a mimic plasmid,in vitro.Cell proliferation was assessed by CCK-8 experiments and the cell cycles were measured by flow cytometry.Cell migration ability was measured by cell wound healing test and transwell test.Cell invasion ability was measured by trans invasion test and the protein change of cell invasion-related protein MMP-2 and MMP-9 were tested by Western blot.In vivo,the growth of tumor in nude mice was further observed to examine whether miR-34a can affect the growth of transplanted tumors in vivo by modulating the expression of SATB2.Results:The expression of miR-34a in OSCC tissues and cell line are lower than para-carcinoma tissues and oral keratin forms cells(P<0.05).After transfecting miR-34a mimic plasmid,the ability of proliferation,metastasis and invasion of Lv-SATB2-HN6 was significantly reduced.Co-transfection of the miR-34a mimics specifically decreased the expression of the SATB2.Conclusions:MiR-34a can down-regulate SATB2 to inhibit the proliferation,migration and invasion of OSCC.

关 键 词：MIR-34A SATB2 口腔鳞癌 HN6细胞 

分 类 号：R739.8[医药卫生—肿瘤]