shRNA靶向干扰<em>COX-2</em>抑制人肝癌裸鼠皮下移植瘤及其血管生成

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中国生物工程杂志 ›› 2011, Vol. 31 ›› Issue (03) : 29-34.

研究报告

shRNA靶向干扰COX-2抑制人肝癌裸鼠皮下移植瘤及其血管生成

陈光辉¹, 刘预², 孔飞飞¹, 邱欣欣¹, 成凤¹, 匡文斌¹, 李朴¹, 涂植光¹

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Inhibitory Effects of Cyclooxygenase-2 Inhibited by shRNA on the Growth and Angiogenesis of Human Liver Cancer Cell Subcutaneous Xenograft Tumors in Nude Mice

CHEN Guang-hui¹, LIU Yu², KONG Fei-fei¹, QIU Xin-xin¹, CHENG Feng¹, KUANG Wen-bin¹, LI Pu¹, TU Zhi-guang¹

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摘要

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。

Abstract

Objective: To investigate the inhibitory effects of recombinant interference plasmid pshRNA-COX-2 on the growth and angiogenesis of human liver cancer cell Hep3B subcutaneous xenograft tumors in nude mice. Methods: The COX-2 mRNA and protein expressions were measured by reverse transcription polymerase chain reaction (RT-PCR) and Western blot, the VEGF mRNA expression was detected by RT-PCR after Hep3B cells were transfected with plasmid pshRNA-COX-2 and selected. Selected Hep3B cells were transplanted into the subcutaneous tissue of nude mice. Xenograft tumor volume was measured every three days and growth curves of tumors were drawn. The mice were killed after four weeks. The expression of COX-2 protein and microvessel density (MVD) in xenograft tumors were observed by immunohistochemistry. Results: Compared to the Hep3B cells without transfected by plasmid pshRNA-COX-2, the inhibition rates of COX-2 mRNA and protein expressions were 65.3% and 52.8% respectively in the pshRNA-COX-2 group (P<0.05), the inhibition rate of VEGF mRNA expression was 56.5% in the pshRNA-COX-2 group (P<0.05). The tumor volume in the pshRNA-COX-2 group was apparently smaller than that of the pshRNA-HK group and the untreated group (P<0.01). The COX-2 score and MVD in the pshRNA-COX-2 group were significantly lower than that of the pshRNA-HK group and the untreated group (P<0.01). Conclusion: The plasmid pshRNA-COX-2 can significantly inhibit the growth and angiogenesis of human liver cancer cell Hep3B subcutaneous xenografts in nude mice through inhibiting COX-2 expression.

导出引用

陈光辉, 刘预, 孔飞飞, 邱欣欣, 成凤, 匡文斌, 李朴, 涂植光. shRNA靶向干扰COX-2抑制人肝癌裸鼠皮下移植瘤及其血管生成[J]. 中国生物工程杂志, 2011, 31(03): 29-34

CHEN Guang-hui, LIU Yu, KONG Fei-fei, QIU Xin-xin, CHENG Feng, KUANG Wen-bin, LI Pu, TU Zhi-guang. Inhibitory Effects of Cyclooxygenase-2 Inhibited by shRNA on the Growth and Angiogenesis of Human Liver Cancer Cell Subcutaneous Xenograft Tumors in Nude Mice[J]. China Biotechnology, 2011, 31(03): 29-34

中图分类号： R735.7

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