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目的:探讨Sam68对急性淋巴细胞白血病Jurkat细胞增殖的影响。方法:针对Sam68 mRNA的第531~552靶位点,构建p LKO-Tet-On载体,制备慢病毒并感染、筛选Jurkat细胞,诱导干扰载体表达;采用real-time PCR和Western blot技术验证干扰效率;用体外细胞集落形成实验观察干扰Sam68基因表达后对细胞集落形成能力的影响;用流式细胞术检测分析Jurkat细胞
Objective: To investigate the effect of Sam68 on the proliferation of acute lymphoblastic leukemia Jurkat cells. Methods: The pLKO-Tet-On vector was constructed based on the target site of Sam68 mRNA from 531 to 552. The lentivirus was constructed and infected. The Jurkat cells were selected and induced to interfere with the expression of the vector. Real-time PCR and Western blot were used to verify the interference efficiency . The effect of interfering with the expression of Sam68 gene on colony forming ability was observed by in vitro colony formation assay. Flow cytometry was used to detect Jurkat cells