CAJ | 학술논문

目的探讨核糖核酸(RNA)在基因水平上干涉血小板源性生长因子(PDGF)的转录与表达,从而抑制视网膜色素上皮细胞(RPE)增生的机制,以期对外伤性PVR的防治提供帮助.方法针对PDGF基因的小干扰核糖核酸(siRNA)序列设计引物,体外转录、转染细胞,寻找具有高效干涉作用的siRNA序列,利用构建具有体内转录siRNA功能的慢病毒载体转染培养的RPE细胞,采用生长曲线及噻唑蓝比色法(MTT)检测PDGF siRNA-慢病毒抑制RPE细胞增生的作用,并用流式细胞仪检测细胞周期的变化.结果获得具有高效干涉PDGF表达的siRNA序列,构建出转录该siR-NA序列的慢病毒载体,且转染RPE细胞后可抑制RPE细胞增生.结论利用RNA的干涉作用,在基因水平上抑制PDGF的转录与表达,从而抑制RPE细胞的增生.
목적 탐토핵당핵산(RNA)재기인수평상간섭혈소판원성생장인자(PDGF)적전록여표체,종이억제시망막색소상피세포(RPE)증생적궤제,이기대외상성PVR적방치제공방조.방법 침대PDGF기인적소간우핵당핵산(siRNA)서렬설계인물,체외전록、전염세포,심조구유고효간섭작용적siRNA서렬,이용구건구유체내전록siRNA공능적만병독재체전염배양적RPE세포,채용생장곡선급새서람비색법(MTT)검측PDGF siRNA-만병독억제RPE세포증생적작용,병용류식세포의검측세포주기적변화.결과 획득구유고효간섭PDGF표체적siRNA서렬,구건출전록해siR-NA서렬적만병독재체,차전염RPE세포후가억제RPE세포증생.결론 이용RNA적간섭작용,재기인수평상억제PDGF적전록여표체,종이억제RPE세포적증생.
Objective To investigate the effect of RNA intererence in transcription and expression of platelet derived growth factor (PDGF) and retinal pigment epithelial cells (RPE) proliferation.Methods According to the PDGF siRNA sequence the primers was designed and siRNA was obtained in vitro by transcription and then transfected cells.The siRNA sequences with high interference effect were looked for.The lentiviral vector with siRNA expression function in vivo was constructed and transfected into cultured RPE cells.The inhibition effect of PDGF siRNA on the proliferation of RPE cells was measured by the growth curve and MTT and the changes of cell cycles were decected by flow cytometry.Results SiRNA sequences with high interference in inhibition of the expression of PDGF were obtained,and the recombinant lentiviral vector of siRNA sequence was constructed.SiRNA-cells which were transfected into RPE cells inhibited the proliferation of RPE cells.Conclusion Using the interference effect of RNA,the transcription and expression of PDGF was inhibited at the gene level,and the proliferation of RPE cells was inhibited.