CAJ | 학술논문

目的：通过体外实验探讨沉默巢蛋白（ Nestin）基因对人食管癌ECA109细胞增殖能力的影响及可能机制。方法合成Lenti-Nestin慢病毒载体，实验设blank组、scrambled组和Lent-Nestin组，转染ECA109细胞，建立稳定沉默Nestin基因的细胞系Lenti-Nestin；用qRT-PCR和Western blot检测Nestin、c-myc和cyclin D1 mRNA和蛋白表达；CCK-8法检测细胞增殖能力；平板集落实验检测细胞的集落形成。结果稳定沉默Nestin的细胞系构建成功；与blank组和scrambled组相比，Lent-Nestin组中Nestin mRNA（ P＜0.01）和蛋白（ P＜0.05）水平明显降低；细胞增殖能力明显降低（ P＜0.01），集落形成能力显著下降（ P＜0.05）；沉默Nestin表达后，c-myc、cyclin D1表达明显受到抑制（ P＜0.05）。结论沉默Nestin基因表达可抑制人食管癌ECA109细胞的增殖能力，其可能通过调控c-myc、cyclin D1表达参与其中。
목적：통과체외실험탐토침묵소단백（ Nestin）기인대인식관암ECA109세포증식능력적영향급가능궤제。방법합성Lenti-Nestin만병독재체，실험설blank조、scrambled조화Lent-Nestin조，전염ECA109세포，건립은정침묵Nestin기인적세포계Lenti-Nestin；용qRT-PCR화Western blot검측Nestin、c-myc화cyclin D1 mRNA화단백표체；CCK-8법검측세포증식능력；평판집락실험검측세포적집락형성。결과은정침묵Nestin적세포계구건성공；여blank조화scrambled조상비，Lent-Nestin조중Nestin mRNA（ P＜0.01）화단백（ P＜0.05）수평명현강저；세포증식능력명현강저（ P＜0.01），집락형성능력현저하강（ P＜0.05）；침묵Nestin표체후，c-myc、cyclin D1표체명현수도억제（ P＜0.05）。결론침묵Nestin기인표체가억제인식관암ECA109세포적증식능력，기가능통과조공c-myc、cyclin D1표체삼여기중。
Objective To investigate the effect of Nestin gene silencing on the proliferation of human esophageal cancer ECA109 cells and possible mechanism .Methods Lenti-Nestin was constructed and transfected into ECA109 cells to establish a stable Nestin-silencing cell line Lenti-Nestin.Blank group , scrambled group , Lenti-Nestin group were set up .The expressions of Nestin , c-myc and cyclin D1 mRNA and protein levels were detected by qRT-PCR and Western blot .The cell proliferation was analyzed by CCK 8 assay .Results The stable Nestin-si-lencing cell line was successfully established .The expression of Nestin mRNA ( P<0.01 ) and protein ( P<0.05 ) levels were reduced significantly and the downregulation evidently suppressed cell proliferation ( P<0.01 ) and col-ony forming capacity ( P<0.05 ) compare with the scrambled group and blank group .However ,The c-myc and cy-clin D1 expression levels in ECA 109 cells in Lenti-Nestin group were significantly lower than that of scrambled group and blank group ( P<0.05 ) .Conclusions Knockdown Nestin expression significantly inhibits the level of esophageal cancer ECA109 cells proliferation,which may act via influencing the expression of c-myc, cyclin D1.