微小RNA Let-7b靶向CD147对大鼠垂体腺瘤GH3细胞侵袭性的影响  

Influence of microRNA Let-7b targeting CD147 on invasion of rat pituitary adenomas GH3 cells

作　　者：谢仕刚 邱鹏[1] 杨伟[1] 张玉宝[1] 徐同江[1] 徐广明[1] 

机构地区：[1]山东大学附属省立医院神经外科,济南250014

出　　处：《中华实验外科杂志》2015年第9期2096-2098,共3页Chinese Journal of Experimental Surgery

基　　金：山东省科技发展计划资助项目（2010GSF10225）;济南市科技发展计划资助项目（201303038）

摘　　要：目的 探寻微小RNA Let-7b（Let-7b）在大鼠垂体腺瘤GH3细胞中的靶基因,并观察其对GH3细胞侵袭性生长的影响.方法 利用TargetScan、PicTar、MiRanda 3种不同的生物信息学方法对靶基因进行筛选;设计Let-7b模拟物（Let-7b mimic）及反义寡核苷酸链序列（AMO）,采用慢病毒转染,分别外源性导入Let-7b mimic以增加Let-7b的表达量、AMO互补结合以阻碍Let-7b功能实现,转染不含有大鼠同源序列的空白病毒为对照组.实时荧光定量核酸扩增检测系统（FQ-PCR）检测各转染组Let-7b表达量的改变;Westen blot检测转染后各组GH3细胞中细胞外基质金属蛋白酶诱导剂（CD147）、基质金属蛋白酶-9（MMP-9）蛋白表达量的改变;Transwell细胞小室侵袭实验研究各组细胞侵袭能力的改变.结果 大鼠GH3细胞中,Let-7b与CD147存在直接的靶向关系,并对CD147、MMP-9蛋白表达及细胞的侵袭能力有明确的抑制作用.转染mimic组的GH3细胞Let-7b的表达量是空白对照组的（9.56±0.51）倍,CD147、MMP-9蛋白表达降低,细胞的侵袭性生长受到抑制,差异有统计学意义（P＜0.05）;转染AMO组的GH3细胞Let-7b的表达为对照组的（0.31±0.01）倍,CD147、MMP-9蛋白表达量增高,细胞的侵袭能力增强,差异有统计学意义（P＜0.05）;对照组与自然生长组Let-7b的表达量、CD147、MMP-9蛋白表达量及细胞侵袭能力差异均无统计学意义（P＞0.05）.结论 Let-7b靶向CD147对GH3细胞的侵袭性有抑制作用.Objective To investigate target genes of microRNA Let-7b in GH3 cells of rat,and study the influence of Let-7b on invasive growth of GH3 cells.Methods Three different bioinformatic ways including TargetScan,PicTar and MiRanda were used to identify the target genes.The Let-7 b mimic and anti-miRNA oligonuclotide （AMO） were designed and transfected into GH3 cells via lentivirus,and the GH3 cells which transfected with vehicle were used as the control group.Real-time quantitative PCR detecting system （FQ-PCR） was used to detect the expression of Let-7b after different transfection.Westen blotting was used to detect the expression of extracellular metalloproteinase inducer （CD147） and matrix metalloprotein-9 （MMP-9）.Transwell experiments were used to detect the invasive ability.Results Let-7b directly targeting CD147 as an inhibitor,suppressed the protein expression of CD147 and MMP-9,and inhibited invasive ability of GH3 cells.After transfection with Let-7b mimic,the mRNA level of Let-7b was 9.56 ± 0.51 times higher than that in the control group （P ＜ 0.05）,the protein expression of CD147 and MMP-9 were reduced,the invasive growth of GH3 cells were inhibited.Inhibiting the Let-7b expression by transfection with AMO,the mRNA level of Let-7b was 0.31 ±0.01 times lower than that in the control group （P ＜ 0.05）,the protein expression of CD147 and MMP-9 were increased,the ability of invasive growth was enhanced.There were no significant differences in the expression of Let-7b,the protein expression of CD147 and MMP-9,and invasive ability of GH3 cells between the control group and the natural growth group.Conclusion Let-7b targeting CD147 inhibits the invasive growth of GH3 cells,it may be used as drug target for gene treatment and clinical prognostic indices.

关 键 词：垂体腺瘤 微小RNA Let-7b 细胞外基质金属蛋白酶诱导剂 基质金属蛋白酶-9 

分 类 号：R735.7[医药卫生—肿瘤]