siRNA沉默TEM8基因对非小细胞肺癌A549细胞增殖和侵袭能力的影响

Effect of siRNA targeting TEM8 gene on proliferation and invasion of non-small cell lung cancer A549 cells

目的探讨RNA干扰靶向抑制TEM8基因的表达对人非小细胞肺癌(non-small cell lung cancer,NSCLC)A549细胞增殖和侵袭能力的的影响。方法采用RT-PCR检测TEM8在5对NSCLC与癌旁正常组织中的表达;采用化学合成针对TEM8基因的小干扰RNA(TEM8 siRNA)下调A549细胞中该基因的表达,应用RT-PCR和Western blot法检测转染后细胞内TEM8的表达,通过Transwell侵袭实验检测TEM8对A549细胞侵袭能力的影响;CCK8实验及平板克隆形成实验检测TEM8对A549细胞增殖能力的影响。结果与癌旁正常肺组织相比,TEM8在NSCLC组织中显著高表达(P<0.05),转染TEM8特异性siRNA后,A549细胞的增殖和侵袭能力均受显著抑制。结论 TEM8在NSCLC中呈过表达,干扰TEM8表达可明显抑制NSCLC的增殖与侵袭能力,推测TEM8可能成为治疗NSCLC的新靶点。

Purpose To investigate the effect of silencing tumor endothelial marker 8 （TEMS） gene expression on proliferation and in- vasion of non-small cell lung cancer （NSCLC） A549 cell. Methods RT-PCR was used to detect and compare the expression of TEM8 in 5 pairs of NSCLC and its adjacent normal lung tissue. Chemically synthesized siRNA targeting TEM8 gene was transfected into the A549 ceils with Lipofectamine 2000. The mRNA and protein expressions of TEM8 were evaluated by RT-PCR and Western blot, re- spectively. The effect of TEM8 silencing on the invasion of A549 cells was evaluate by transwell invasion assay. The influence of TEM8 on A549 cell proliferation was detected by CCK8 assay and plate colony formation assay. Results Compared with adjacent normal lung tissues, TEM8 in NSCLC was significantly up-regulated （ P 〈 0. 05 ）. Knockdown of TEM8 by siRNA inhibited A549 cell proliferation and invasion. Conclusion Over-expressed of TEM8 in NSCLC and the proliferation and invasion capability of NSCLC was inbibited with TEM8 siRNA, which suggest TEM8 may be a new therapic target for NSCLC.