摘要
目的采用siRNA干扰技术沉默人非霍奇金淋巴瘤Raji细胞中Pokemon基因,观察Raji细胞增殖活性的变化,并探讨其可能的分子机制。方法构建靶向Pokemon基因的siRNA重组慢病毒载体并转染Raji细胞。采用实时定量PCR法和Western blotting法检测Raji细胞Pokemon基因的沉默效果,在Raji细胞中沉默Pokemon基因的表达后采用实时定量PCR法和Western blotting法检测bcl-6和突变型p53表达水平的变化;采用流式细胞术检测Pokemon基因沉默后Raji细胞凋亡的情况。结果利用Pokemon靶向siRNA重组慢病毒载体感染Raji细胞有效沉默Raji细胞Pokemon基因表达(P<0.05)。沉默Pokemon基因后,bcl-6和突变型p53基因和蛋白表达均显著降低(P<0.05),细胞凋亡率增高(P<0.05)。结论 siRNA沉默Pokemon基因有效降低了人非霍奇金淋巴瘤Raji细胞的bcl-6和突变型p53基因和蛋白表达,促进细胞凋亡,抑制其增殖。Pokemon基因有望成为非霍奇金淋巴瘤治疗的新靶点。
Objective Small interference RNA（siRNA） was used to silence Pokemon gene in human non-Hodgkin’s lymphoma Raji cells,then to observe the change of proliferation of Raji cells and explore its possible molecular mechanisms. Methods Pokemon-targeted siRNA was constructed with lentivirus vector and transfected to Raji cells. RT-PCR and Western blotting were adopted to confirm the silence effect of Pokemon gene in Raji cells,which were then used to determine the mRNA and protein expressions of bcl-6 and mutant p53. Flowcytometry was used to detect the cell apoptosis of each group. Results Pokemon-siRNA constructed with lentivirus vector could efficiently silence the expression of Pokemon gene in Raji cell（P ＆lt; 0. 05）. After that,the mRNA and protein expressions of bcl-6 and mutant p53 were significantly decreased（P ＆lt; 0. 05） and the cell apoptosis rate was markedly elevated compared with the controls. Conclusion The siRNA Pokemon gene silencing promotes human non-Hodgkin’s lymphoma Raji cells apoptosis by lowering the expressions of bcl-6 and mutant p53 gene and protein and inhibiting the proliferation. Pokemon gene is expected to become a newtarget for non-Hodgkin’s lymphoma treatment.
出处
《山东大学学报（医学版）》
CAS
北大核心
2014年第5期53-57,共5页
Journal of Shandong University:Health Sciences
基金
山东省科技攻关计划(2008GG10002049)
山东省优秀中青年科研奖励基金(BS2009YY049)
