重庆医学
重慶醫學
중경의학
CHONGQING MEDICAL JOURNAL
2015年
16期
2170-2173
,共4页
微RNA-335%rho相关激酶类1%骨肉瘤%基因表达
微RNA-335%rho相關激酶類1%骨肉瘤%基因錶達
미RNA-335%rho상관격매류1%골육류%기인표체
microRNA-335%rho-associated kinases 1%osteosarcoma%gene 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。
目的：探討miR‐335及rho相關捲麯螺鏇形成蛋白激酶1（ROCK1）在骨肉瘤組織及細胞繫中錶達的情況及兩者的相關性。方法臨床選取18例骨肉瘤組織標本及配對正常骨骼肌組織標本；設定人正常成骨細胞為對照組，骨肉瘤細胞繫M G‐63和U2‐OS為實驗組，RT‐PCR檢測骨肉瘤組織與各組細胞中 miR‐335與 ROCK1 mRNA 的錶達差異；miR‐335 mimic轉染MG‐63和U2‐OS兩細胞繫，檢測miR‐335過錶達對 ROCK1 mRNA和蛋白錶達的影響。結果 miR‐335在骨肉瘤組織及細胞繫中特異性低錶達，而ROCK1則呈高錶達，兩者呈負相關；轉染miR‐335 mimic後，miR‐335 mRNA過錶達，ROCK1 mRNA和蛋白錶達在MG‐63和U2‐OS兩細胞繫中受到顯著抑製。結論 miR‐335與ROCK1錶達在骨肉瘤組織學水平和細胞學水平均成負相關；過錶達miR‐335可降低ROCK1的錶達。
목적：탐토miR‐335급rho상관권곡라선형성단백격매1（ROCK1）재골육류조직급세포계중표체적정황급량자적상관성。방법림상선취18례골육류조직표본급배대정상골격기조직표본；설정인정상성골세포위대조조，골육류세포계M G‐63화U2‐OS위실험조，RT‐PCR검측골육류조직여각조세포중 miR‐335여 ROCK1 mRNA 적표체차이；miR‐335 mimic전염MG‐63화U2‐OS량세포계，검측miR‐335과표체대 ROCK1 mRNA화단백표체적영향。결과 miR‐335재골육류조직급세포계중특이성저표체，이ROCK1칙정고표체，량자정부상관；전염miR‐335 mimic후，miR‐335 mRNA과표체，ROCK1 mRNA화단백표체재MG‐63화U2‐OS량세포계중수도현저억제。결론 miR‐335여ROCK1표체재골육류조직학수평화세포학수평균성부상관；과표체miR‐335가강저ROCK1적표체。
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 .
