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siRNA抑制食管癌EC9706细胞CXCR4基因表达的实验

Initial on Expression of CXCR4 mRNA in EC9706 Cell Silenced by siRNA

  • 摘要: 目的运用RNAi技术沉默食管癌EC9706细胞CXCR4(chemokine receptor4)基因表达,观察其对肿瘤细胞的生长和转移影响。方法设计CXCR4 基因为靶向的siRNA, 构建siRNA表达载体,通过脂质体将siRNA表达载体转入EC9706细胞。荧光定量PCR法检测细胞CXCR4 mRNA表达水平;流式细胞术检测细胞周期;MTT法检测细胞侵袭和增殖的情况。结果构建出CXCR4 基因为靶向的siRNA表达载体pRNAT-U6.2/Lenti-siCX1和pRNAT-U6.2/Lenti -siCX2;荧光定量PCR结果显示,转染pRNAT-U6.2/Lenti-siCX1的EC9706细胞和转染pRNAT-U6.2/Lenti-siCX2的EC9706细胞与对照组相比CXCR4 mRNA的表达水平明显降低(P<0.01);细胞生长速度较对照组明显减慢(P<0.01);流式细胞仪检测结果显示:转染pRNAT-U6.2/Lenti-siCX1的EC9706S期细胞比例低于对照组(P<0.05)。结论沉默CXCR4基因表达对食管癌EC9706细胞的生长和侵袭、转移有明显的抑制作用。

     

    Abstract: Objective To study the inhibitory effect of small interfering RNA(siRNA) targeting CXCR4 gene in EC9706 cells. Methods We designed two siRNAs against CXCR4 gene.Two siRNAs were transfected into EC9706 cells with lipofectamine,respectively.The expression level of CXCR4 mRNA in each group was detected by real-time PCR ; the effect of CXCR4 on the cell cycle was observed by flow cytometry; the effect of CXCR4 on the growth of the cells was observed by MTT method. Results Compared with empty vector control group and untransfected group, the mRNA level of CXCR4 in siRNA transfected group was obviously reduced,and the growth speed of cells was lower, S phase proportion cut down in EC9706 cells transfected with pRNAT-U6.2/Lenti- siCX1,the exercise performance of the cells cut down in siRNA transfected group cells. Conclusion RNA interference can effectively inhibit CXCR4 expression, which plays an important role in inhibiting the growth and metastasis of the tumor.

     

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