RNA干扰LMP1阻滞淋巴瘤细胞于G0/G1期并提高细胞对顺铂的敏感性

LMP1 Knockdown Retards G0/G1 Phase and Increases Chemosensitivity to Cisplatin in Lymphoma B95.8 Cell Line

目的利用RNA干扰方法下调EB病毒潜伏膜蛋白1(latent membrane protein,LMP1)的表达,探讨LMP1对化疗药物敏感性的影响。方法采用小分子干扰片段转染EB病毒阳性淋巴瘤B95.8细胞,RT-PCR和Western blot分别检测LMP1的mRNA和蛋白的表达水平;联合应用化疗药物顺铂,通过MTT法观察细胞增殖活性变化。结果干扰LMP1后,其mRNA和蛋白的表达均明显下调;细胞增殖能力减弱;细胞被阻滞在G0/G1期;并且细胞对顺铂的IC50明显降低。结论干扰LMP1提高了B95.8细胞对化疗药物的敏感性,揭示LMP1可能是一个潜在的药物靶点。

Objective This study was designed to investigate the effect of LMP1 knockdown on chemosensitivity of lymphoma B95.8 cells. Methods Small interference RNA （siRNA） for LMP1 was designed and transfected to B95.8 cells. LMP1 mRNA and protein expression levels were detected by RT-PCR and Western blot, respectively. Cell cycle was assessed by flow cyto,netry and the sensitivity of B95-8 cells to cisplatin was determined by MTT cytotoxicity method. Results The designed siRNA was effective in reducing the expression of LMP1 protein. LMP1 knockdown resulted in G0/G1 phase cell cycle arrest, and the value of IC50 of cisplatin on B95.8 cells was also decreased. Conclusion Chemosensitivity of B95.8 cells to cisplatin was increased in cells with LMP1 knockdown, suggesting that LMP1 may be a target for chmotherapy.