Wang Liguo, Wu Guojun, Yu Lei, Yuan Jianlin, Fang Fang, Zhai Zhenbo, Wang Fuli, Wang He
Department of Urology, Xijing Hospital, Fourth Military Medical University, Xi'an City, China.
Cancer Biol Ther. 2006 Jun;5(6):608-14. doi: 10.4161/cbt.5.6.2661. Epub 2006 Jun 5.
CD147, also named extracelluar matrix metalloproteinase inducer (EMMPRIN), has been proved to be involved in the invasion and metastasis processes of tumor cells in many types of cancers. To determine the role of CD147 in the invasiveness properties of prostate cancer, we successfully downregulated CD147 by RNA interference (RNAi) technology, in PC-3 cell line at high level of CD147 expression. PC-3 cells were transfected with a pSilencer 4.1-CMV neo Vector coding for an RNA composed of two identical 19-nucleotide sequence motifs in an inverted orientation, separated by a 9-bp spacer to form a hairpin dsRNA capable of mediating target CD147 inhibition. Gelatin zymography was employed to determine the effect on reducing secretions of MMP-2 and MMP-9 of the transfected cells. Matrigel invasion assay was performed to evaluate the invasion ability of PC-3 cells in vitro. Our results showed that CD147 expression was significantly inhibited by small interfering RNAs (siRNA) transfectants in PC-3 cells at mRNA and protein levels, which resulted in dramatic reduction of invasion ability in tumor cells. Moreover, downregulation of CD147 resulted in reducing secretions of MMP-2, MMP-9. Taken together, CD147 downregulation by RNAi technology decreases the invasive capability of prostate cancer cells, demonstrating that stable expression of siRNA CD147 could potentially be an experimental approach for prostate cancer gene therapy.
CD147,也被称为细胞外基质金属蛋白酶诱导因子(EMMPRIN),已被证明在多种癌症的肿瘤细胞侵袭和转移过程中发挥作用。为了确定CD147在前列腺癌侵袭特性中的作用,我们利用RNA干扰(RNAi)技术,在CD147高表达的PC-3细胞系中成功下调了CD147的表达。用编码由两个反向排列的相同19个核苷酸序列基序组成的RNA的pSilencer 4.1-CMV neo载体转染PC-3细胞,这两个基序由一个9碱基对的间隔区隔开,形成能够介导对靶标CD147抑制作用的发夹状双链RNA。采用明胶酶谱法测定转染细胞对MMP-2和MMP-9分泌的影响。进行基质胶侵袭试验以评估PC-3细胞的体外侵袭能力。我们的结果表明,小干扰RNA(siRNA)转染剂在mRNA和蛋白质水平上显著抑制了PC-3细胞中CD147的表达,这导致肿瘤细胞的侵袭能力显著降低。此外,CD147的下调导致MMP-2、MMP-9分泌减少。综上所述,通过RNAi技术下调CD147可降低前列腺癌细胞的侵袭能力,表明siRNA CD147的稳定表达可能是前列腺癌基因治疗的一种实验方法。