现代肿瘤医学
現代腫瘤醫學
현대종류의학
JOURNAL OF MODERN ONCOLOGY
2015年
1期
15-19
,共5页
MicroRNA-100%mTOR%上皮间质转化
MicroRNA-100%mTOR%上皮間質轉化
MicroRNA-100%mTOR%상피간질전화
MicroRNA-100%mTOR%EMT
目的:探讨microRNA-100调节mTOR表达对肝癌细胞侵袭转移及上皮间质转化的影响并研究其可能的分子机制。方法:通过脂质体介导将microRNA-100模拟物及阴性对照转染HepG2细胞,将细胞分为转染试剂对照组( mock)、阴性对照组( control)和microRNA-100 mimic( miRNA-100 mimic)组。采用real-time PCR检测转染后细胞中miRNA-100的表达水平,Western blot检测mTOR的表达变化,细胞划痕实验检测细胞的迁移能力,Tranwell检测细胞的侵袭能力,Western blot检测细胞中MMP-2、MMP-9、N-cadherin、Vimentin、α-SMA和E-cadherin的表达。结果:过表达miRNA-100通过抑制mTOR的表达抑制肝癌细胞的迁移及侵袭能力,并通过下调肝癌细胞中MMP-2、MMP-9、N-cadherin、Vimentin和α-SMA的表达,上调E-cadherin的表达,阻抑上皮细胞-间质转化( epithelial-mesenchymal transition,EMT)过程的发生。结论:MicroRNA-100可能通过降低mTOR的表达抑制肝癌细胞的转移侵袭能力,并抑制上皮间质转化过程的发生。
目的:探討microRNA-100調節mTOR錶達對肝癌細胞侵襲轉移及上皮間質轉化的影響併研究其可能的分子機製。方法:通過脂質體介導將microRNA-100模擬物及陰性對照轉染HepG2細胞,將細胞分為轉染試劑對照組( mock)、陰性對照組( control)和microRNA-100 mimic( miRNA-100 mimic)組。採用real-time PCR檢測轉染後細胞中miRNA-100的錶達水平,Western blot檢測mTOR的錶達變化,細胞劃痕實驗檢測細胞的遷移能力,Tranwell檢測細胞的侵襲能力,Western blot檢測細胞中MMP-2、MMP-9、N-cadherin、Vimentin、α-SMA和E-cadherin的錶達。結果:過錶達miRNA-100通過抑製mTOR的錶達抑製肝癌細胞的遷移及侵襲能力,併通過下調肝癌細胞中MMP-2、MMP-9、N-cadherin、Vimentin和α-SMA的錶達,上調E-cadherin的錶達,阻抑上皮細胞-間質轉化( epithelial-mesenchymal transition,EMT)過程的髮生。結論:MicroRNA-100可能通過降低mTOR的錶達抑製肝癌細胞的轉移侵襲能力,併抑製上皮間質轉化過程的髮生。
목적:탐토microRNA-100조절mTOR표체대간암세포침습전이급상피간질전화적영향병연구기가능적분자궤제。방법:통과지질체개도장microRNA-100모의물급음성대조전염HepG2세포,장세포분위전염시제대조조( mock)、음성대조조( control)화microRNA-100 mimic( miRNA-100 mimic)조。채용real-time PCR검측전염후세포중miRNA-100적표체수평,Western blot검측mTOR적표체변화,세포화흔실험검측세포적천이능력,Tranwell검측세포적침습능력,Western blot검측세포중MMP-2、MMP-9、N-cadherin、Vimentin、α-SMA화E-cadherin적표체。결과:과표체miRNA-100통과억제mTOR적표체억제간암세포적천이급침습능력,병통과하조간암세포중MMP-2、MMP-9、N-cadherin、Vimentin화α-SMA적표체,상조E-cadherin적표체,조억상피세포-간질전화( epithelial-mesenchymal transition,EMT)과정적발생。결론:MicroRNA-100가능통과강저mTOR적표체억제간암세포적전이침습능력,병억제상피간질전화과정적발생。
Objective:To investigate the effect of microRNA-100 on the migration,invasion and epithelial-mes-enchymal transition( EMT)process of human hepatocarcinoma HepG2 cells and the possible mechanism. Methods:Synthetic miRNA-100 mimic and its negative control were transfected into HepG2 cells by liposome method. We di-vided cells into three groups:mock group,control group and miRNA-100 mimic group. After transfection,the ex-pression of mTOR at mRNA level was detected by quantitative real-time PCR. The migration of HepG2 cells was de-tected by wound scratch assay,and the invasion ability changes were analyzed in transwell. Expressions of MMP-2, MMP-9,N-cadherin,Vimentin,α-SMA and E-cadherin were analyzed by Western blot. Results:MiRNA-100 can inhibit the migration and invasion ability via down -regulate the mTOR expression. The Western blot results showed that miRNA-100 could down-regulate the expression of MMP-2,MMP-9,N-cadherin,Vimentin and α- SMA and up - regulated the expression of E - cadherin,and inhibit the progress of EMT. Conclusion:MiRNA -100 can suppress the migration and invasion process and EMT process,it is may be achieved by down - regulating mTOR gene expression.