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Effects of silencing cyclooxygenase-2 expression via RNA interference on the tumorigenicity of the SMMC-7721 human hepatocarcinoma cell line

机译:RNA干扰沉默环氧合酶-2表达对SMMC-7721人肝癌细胞系致瘤性的影响

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摘要

We constructed a vector carrying a shRNA sequence against cyclooxygenase-2 (COX-2) that was subsequently transfected into the human hepatocarcinoma cell line SMMC-7721. Furthermore, we established a COX-2-deficient stable cell line and a model of tumor-shRNA transplantation in nude mice. Negative shRNA was used as the control. The tumor volume in the experimental group was smaller compared to that in the control group. Hematoxylin and eosin staining indicated that the cells in the experimental group differentiated better than those in the control group. The COX-2 mRNA level in the tumor tissues injected with SMMC-7721/COX-2i was markedly downregulated compared to that in the tumor tissues injected with SMMC-7721/Negative shRNA. The inhibition rate reached 68.6%. Immunohistological study showed a significantly strong COX-2 expression in the control group tumor cells, whereas the experimental group exhibited moderate expression, indicating the inhibition of COX-2 expression after transfection of cells with shRNA against COX-2. Western blot analysis further proved the inhibition of COX-2 expression. In conclusion, RNAi-mediated regulation of COX-2 expression could efficiently inhibit liver-transplanted tumor growth in BALB/c nude mice.
机译:我们构建了一个带有针对环氧合酶2(COX-2)的shRNA序列的载体,随后将其转染到人肝癌细胞系SMMC-7721中。此外,我们建立了一个COX-2缺陷稳定细胞系和裸鼠中的肿瘤shRNA移植模型。阴性shRNA用作对照。实验组的肿瘤体积比对照组小。苏木精和曙红染色表明,实验组的细胞分化优于对照组。与注射SMMC-7721 /阴性shRNA的肿瘤组织相比,注射SMMC-7721 / COX-2i的肿瘤组织中的COX-2 mRNA水平显着下调。抑制率达到68.6%。免疫组织学研究显示,对照组肿瘤细胞中COX-2表达明显增强,而实验组则表现出中等表达,表明在用shRNA转染COX-2细胞后,COX-2表达受到抑制。蛋白质印迹分析进一步证明了对COX-2表达的抑制。总之,RNAi介导的COX-2表达调节可有效抑制BALB / c裸鼠肝移植肿瘤的生长。

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