中文摘要：

目的 探讨反义微小RNA-21寡核苷酸（antisense oligonucleotide micro RNA-21,AS-miRNA-21）抑制Tb3.1人舌鳞状细胞癌增殖的效果和机制.方法 实验分3组,①空白对照组 ②无义寡核苷酸转染组 ③AS-miRNA-21转染组.寡核苷酸介导转染反义寡核苷酸敲低Tb3.1细胞miRNA-21表达.使用荧光实时定量聚合酶链反应（PCR）鉴定转染后Tb 3.1细胞miRNA-21表达水平 甲基噻唑基四唑（MTT）法检测转染后Tb 3.1细胞生存率 流式细胞术检测转染后Tb3.1早期凋亡 Matrigel基质生长实验检测转染后Tb 3.1细胞生长形成球形集落能力 Transwell体外迁移实验检测转染后Tb 3.1细胞迁移能力 蛋白质印迹法检测转染后Tb3.1细胞增殖核抗原（antigen KI-.67,Ki67）、B细胞淋巴瘤2（B cell lymphoma 2,Bcl-2）、人第10号染色体缺失的磷酸酶及张力蛋白同源的基因（phosphatase and tensin homolog,PTEN）、基质金属蛋白酶2、9（matrix metalloproteinase 2/9,MMP-2、MMP-9）和组织基质蛋白酶抑制因子蛋白1（tissue inhibitor of metalloproteinase 1,TIMP-1）蛋白表达.结果 荧光实时定量PCR显示转染后miRNA-21表达水平下调 转染第4天,AS-miRNA-21转染组肿瘤细胞生长速度[（53.43±11.83）%]低于其他两组[（91.32±8.02）%和100%]（F=27.02,P=0.00） 细胞凋亡率显著升高[（12.23±2.92）%,F=26.641,P=0.001] AS-miRNA-21转染组细胞生长不能形成球形克隆且通过Transwell小室聚碳酸酯膜的细胞数小于空白对照组（F=268.231,P=0.000） Ki67、Bcl-2、MMP-2和MMP-9蛋白表达下调,PTEN和TIMP-1蛋白表达上调.结论 敲低miRNA-21后Tb3.1人舌癌细胞增殖与侵袭能力被抑制,并为探索miRNA-21调控人舌癌发生机制提供实验依据.

英文摘要：

Objective To investigate the effect of micro RNA-21 （miRNA-21） knocking on the Tb3.1 human tongue squamous cell carcinoma growth. Methods Anti-sense miRNA-21 oligonucleotide was delivered with oligofectamine to suppress Tb 3. 1 tongue cancer cell growth in vitro. Real-time polymerase chain reaction （PCR） was conducted to detect the miRNA-21 expression after transfection. Methyl thiazolyl tetrazolium（MTT） assay was used to determine Tb 3. 1 cell survival rate. Apoptosis were examined by flowcytometry. Matrigel matrix and transwell assay were used to determine Tb 3.1 cell colony formation and migration ability. Antigen KI-67 （Ki67）, B cell lymphoma （Bcl-2）, phosphatase and tensin homolog （PTEN）, matrirx metalloproteinase 2（MMP-2, MMP-9） and tissue inhibitor of metalloproteinase 1 （TIMP-1） protein expression in Tb 3. 1 cell were measured by Western blotting. Results miRNA-21 expression was decreased in miRNA-21 antisense oligonucleotide （ASODN） group. The survival rate of Tb 3. 1 cells with AS-miRNA-21 transfection was significantly suppressed （F=27.02, P = 0.00） and early phase apoptosis（F =26. 641 ,P = 0. 001） induced in Tb 3.1 cell. Ki67, Bcl-2, MMP-2 and MMP-9 protein weredown regulated while PTEN and TIMP-1 protein expression was increased. Conclusions Blocking miRNA-21 expression in Tb3.1 cell could suppress cancer cell growth in vitro and miRNA-21 can serve as a novel target candidate for human tongue cancer gene therapy.