慢病毒介导的RNA干扰抑制白血病细胞增殖的应用

Application of the lentiviral vector mediated RNAi to inhibit the proliferation of leukemia cell

通过慢病毒载体介导的RNA干扰(RNAi)技术,沉默成人T细胞白血病(ATL)中关键的病毒基因HBZ,并研究RNA干扰后对白血病细胞增殖的影响.首先根据HBZ基因序列设计RNA干扰片段,构建其慢病毒载体,并将其包装成病毒.然后用HBZ siRNA慢病毒感染白血病细胞株TL-Om1,通过RT-PCR和Western blot检测HBZ基因和蛋白表达情况,MTT技术检测沉默HBZ后TL-Om1细胞的增殖情况,利用RT-PCR检测病毒感染后下游生长相关基因的表达.实验结果表明:利用慢病毒介导的RNA干扰技术能有效降低HBZ的表达,进而抑制了白血病细胞的增殖;沉默HBZ后,细胞内E2F1,PCNA和Myc等下游生长相关基因的表达受到了明显抑制.这将为慢病毒介导的RNA干扰技术应用于临床治疗成人T细胞白血病提供重要的实验依据.

Lentiviral vector-mediated RNA interference (RNAi) technique was used to silence the critical viral gene HBZ in adult T cell leukemia (ATL), it was investigated the effect of RNA interference on the proliferation of leukemia cells. Firstly, the RNA interference fragment was designed according to the HBZ gene sequences, constructed its lentiviral vector and packed it into the virus. The HBZ siRNA lentiviral was used to infect the TL-Om1, a kind of leukemia cell strains. RT-PCR and Western blot were used to detect the expression of HBZ gene and protein, MTT assay was used to measure the proliferation of TL-Om1 after HBZ silenced and RT-PCR was used to detect the expression of downstream growth related genes after viral infection. The results showed that lentiviral vector-mediated RNA interference could effectively reduce the expression of HBZ and further inhibit the proliferation of leukemia cells. Besides the expression of E2F1 , PCNA, Myc and other downstream growth related genes were inhibited significantly after HBZ silenced. An important experimental evidence was provided for lentivirus mediated RNA interference being applied in clinical treatment of adult T-cell leukemia.