干扰CD147表达抑制白血病细胞SHI-1增殖及移行

RNAi-mediated silencing of CD147 inhibits the proliferation and invasion of leukemic cells SHI-1

目的构建针对细胞外基质金属蛋白酶诱导因子(CD147)的慢病毒干扰载体,探讨干扰CD147表达后对白血病细胞系SHI-1细胞体外增殖、侵袭、转移能力的影响。方法设计针对CD147的干扰序列片段,构建于慢病毒载体pGCSIL-GFP上,重组慢病毒载体与包装载体共转染293T细胞,收集病毒上清,测定病毒滴度。病毒上清感染SHI-1细胞,RT-PCR法检测CD147、MMP-2及MMP-9 mRNA表达,Western blotting法检测CD147蛋白水平;MTT法检测细胞体外增殖能力,体外SHI-1细胞与骨髓基质细胞共培养跨Matrigel基质胶检测SHI-1细胞的体外侵袭能力。结果成功构建慢病毒干扰载体,获得重组慢病毒,滴度达1×109TU/mL。病毒感染SHI-1细胞后,SHI-1/CD147i细胞的CD147 mRNA较感染阴性对照病毒的SHI-1/NC细胞下调86.7%,CD147蛋白表达下降91%,SHI-1/CD147i细胞MMP-2及MMP-9 mRNA表达较SHI-1/NC细胞分别下降78.3%和70.6%。SHI-1/CD147i细胞体外增殖能力较SHI-1/NC及SHI-1细胞明显下降;与骨髓基质细胞共培养24 h后,SHI-1、SHI-1/NC、SHI-1/CD147i细胞移行至Transwell下层的细胞占接种细胞数比例分别为(18.2±2.5)%、(16.5±2.7)%、(4.5±1.2)%,SHI-1/CD147细胞移行能力显著低于SHI-1和SHI-1/NC细胞。结论干扰SHI-1细胞的CD147表达后,通过下调MMPs表达抑制SHI-1细胞体内外增殖及移行能力,CD147可能成为治疗急性白血病的靶点。

of CD147 was detected by Western blotting.The capability of proliferation and infiltration of SHI-1 cell were examined by MTT and trans-matrigel invasion assay co-cultured with leukemia BMSCs in vitro.Results The lentivirus contai-ning the CD147 shRNA was constructed successfully.The titer of recombined virus was 1×109 TU/mL.The mRNA and protein of CD147 in SHI-1/CD147i cells decreased by 86.7%and 91%respectively after the SHI-1 cells were infected by the lentivirus containing the CD147 siRNA.The proliferation capability of SHI-1/CD147i cells significantly decreased compared with that of SHI-1 and SHI-1/NC cells.The mRNA expressions of MMP-2 and MMP-9 in SHI-1/CD147i cells were significantly lower than that in SHI-1/NC and SHI-1 cells.The SHI-1/CD147i cells demonstrated significantly lower invasion rate than SHI-1 cells and SHI-1/NC cells when co-cultured with BMSCs.Conclusion CD147 plays im-portant roles in the leukemia cell proliferation and infiltration.CD147 should be a potential target for the treatment of acute leukemia.