miR-335及ROCK1在骨肉瘤中表达及相互关系研究

摘要

Objective To investigate the expression and correlation of microRNA‐335(miR‐335) and ROCK1 in osteosarco‐ma tissue and osteosarcoma cell lines MG‐63 and U2‐OS .Methods Eighteen paired tissue specimens of osteosarcoma and matched normal tissue were chosen clinically .Two osteosarcoma cell lines MG‐63 and U2‐OS were chosen as experimental group while the human osteoblast cell line hFOB 1 .19 as control group ,Quantitative real‐time PCR were used to evaluate the different expression of miR‐335 and ROCK1 mRNA in osteosarcoma tissue and cell lines;miR‐335 mimic was transfected into MG‐63 and U2‐OS ,while Quantitative real‐time PCR and Western blot were used to evaluate the effect of over‐expressed miR‐335 on ROCK1 mRNA and protein level .Results The result of real‐time PCR indicated a special lower expression of miR‐335 but higher ROCK1 ,and the cor‐relation between miR‐335 and ROCK1 was negative .Western blot showed that miR‐335 could decrease ROCK1 protein expression . miR‐335 mRNA showed a hyper‐expression while lower ROCK1 mRNA and protein after transfection of miR‐335 mimic .Conclusion The expression of miR‐335 and ROCK1 shows a negative correlation in both tissue and cell level;over‐expression of miR‐335 can decrease ROCK1 expression .%目的：探讨miR‐335及rho相关卷曲螺旋形成蛋白激酶1（ROCK1）在骨肉瘤组织及细胞系中表达的情况及两者的相关性。方法临床选取18例骨肉瘤组织标本及配对正常骨骼肌组织标本；设定人正常成骨细胞为对照组，骨肉瘤细胞系M G‐63和U2‐OS为实验组，RT‐PCR检测骨肉瘤组织与各组细胞中 miR‐335与 ROCK1 mRNA 的表达差异；miR‐335 mimic转染MG‐63和U2‐OS两细胞系，检测miR‐335过表达对 ROCK1 mRNA和蛋白表达的影响。结果 miR‐335在骨肉瘤组织及细胞系中特异性低表达，而ROCK1则呈高表达，两者呈负相关；转染miR‐335 mimic后，miR‐335 mRNA过表达，ROCK1 mRNA和蛋白表达在MG‐63和U2‐OS两细胞系中受到显著抑制。结论 miR‐335与ROCK1表达在骨肉瘤组织学水平和细胞学水平均成负相关；过表达miR‐335可降低ROCK1的表达。