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目的:探讨重组干扰质粒pshRNA-COX-2对人肝癌细胞Hep3B裸鼠皮下移植瘤生长和肿瘤血管生成的抑制作用。方法:重组干扰质粒pshRNA-COX-2转染Hep3B细胞并筛选后,RT-PCR和Western blot检测COX-2mRNA和蛋白表达,RT-PCR检测VEGFmRNA表达。将被成功转染的Hep3B细胞种植于裸鼠皮下,测量肿瘤大小,4周后处死裸鼠,免疫组织化学法检测肿瘤组织中COX-2蛋白表达和肿瘤微血管密度(MVD)。结果:与未转染细胞相比,干扰组COX-2mRNA和蛋白表达抑制率分别为65.3%和52.8%(P<0.05),干扰组VEGFmRNA表达抑制率为56.5%(P<0.05)。干扰组瘤体大小明显小于阴性组和空白组(P<0.01)。干扰组COX-2得分和MVD均明显低于阴性组和空白组(P<0.01)。结论:pshRNA-COX-2通过抑制COX-2表达明显抑制人肝癌细胞Hep3B裸鼠皮下移植瘤生长和肿瘤血管生成。
Objective: To investigate the inhibitory effect of recombinant shRNA plasmid pshRNA-COX-2 on the growth and tumor angiogenesis of human hepatocellular carcinoma Hep3B cells in nude mice. METHODS: Hep3B cells were transfected with recombinant plasmid pshRNA-COX-2 and screened for COX-2 mRNA and protein expression by RT-PCR and Western blot. VEGFmRNA expression was detected by RT-PCR. The successfully transfected Hep3B cells were implanted subcutaneously in nude mice and the tumor size was measured. After 4 weeks, the nude mice were sacrificed and the expression of COX-2 protein and tumor microvessel density (MVD) in the tumor tissue were detected by immunohistochemistry. Results: Compared with untransfected cells, the inhibitory rates of COX-2 mRNA and protein expression in the interference group were 65.3% and 52.8%, respectively (P <0.05). The inhibition rate of VEGF mRNA expression in the interference group was 56.5% (P <0.05). The tumor size of the interference group was significantly smaller than that of the negative group and the blank group (P <0.01). The scores of COX-2 and MVD in the interference group were significantly lower than those in the negative group and the blank group (P <0.01). CONCLUSIONS: pshRNA-COX-2 significantly inhibits the growth of subcutaneous xenografts and tumor angiogenesis in human hepatoma Hep3B nude mice by inhibiting COX-2 expression.