shRNA干扰CDX2基因表达对裸鼠结直肠癌肝转移瘤的影响

发布时间：2018-06-26 17:40

  本文选题：干扰尾型同源盒基因 + 短发夹RNA　； 参考：《中国肿瘤生物治疗杂志》2016年05期

【摘要】：目的:建立人结直肠癌裸鼠肝转移瘤模型,探讨干扰尾型同源盒基因2(caudal-related homeobox 2,CDX2)基因表达对结直肠癌肝转移瘤的影响。方法:通过慢病毒载体将CDX2-shRNA体外转染人结直肠癌细胞SW480、HT29,筛选建立稳定干扰CDX2基因表达细胞株和阴性病毒细胞株;将稳定干扰CDX2基因表达的结直肠癌细胞(SW480-KD、HT29-KD)、空白对照细胞(SW480-CON、HT29-CON)和阴性对照细胞(SW480-NC、HT29-NC)接种到裸鼠脾下极包膜内,建立裸鼠结直肠癌肝转移瘤模型,每天称量裸鼠体质量并观察其摄食、活动及精神情况;50 d后处死裸鼠,分别从大体水平以及镜下观察肝转移瘤情况。结果:成功构建裸鼠结直肠癌肝转移模型,H-E染色确认为结直肠癌肝转移。各细胞干扰组(SW480-KD、HT29-KD)裸鼠体质量明显低于相应空白对照组和阴性对照组[SW480-KD:(15.02±2.00)vs(18.00±2.48)、(18.03±2.05)g;F=3.761,P0.05;HT29-KD:(15.39±2.16)vs(17.96±2.48)、(18.30±1.87)g;F=3.721,P0.05]。SW480-KD组肝转移瘤数目明显多于SW480-CON组和SW480-NC组[(57.83±22.56)vs(29.50±16.90)、(28.20±15.40)个;F=4.197,P0.05];HT29-KD组肝转移瘤数目明显多于HT29-CON组和HT29-NC组[(56.83±29.16)vs(26.40±12.76)、(21.00±11.50)个;F=4.467,P0.05)]。结论:脾注射法裸鼠肝转移模型可作为体内研究基因功能的理想模型;干扰CDX2基因表达可促进结直肠癌SW480和HT29细胞在体内的转移。
[Abstract]:Objective: to establish a nude mouse liver metastasis model of human colorectal cancer and to investigate the effect of interfering tail type homologous box gene 2 (caudal-related homeobox 2 + CDX2) gene expression on liver metastasis of colorectal cancer. Methods: CDX2-shRNA was transfected into human colorectal cancer cell line SW480 (HT29) by lentivirus vector in vitro, and stable interfering CDX2 gene expression cell line and negative virus cell line were established. Colorectal cancer cells (SW480-KDfHT29-KD), blank control cells (SW480-CONNHT29-CON) and negative control cells (SW480-NCChHT29-NC), which interfered with CDX2 gene expression, were inoculated into the subsplenic polar capsule of nude mice to establish nude mice liver metastases model of colorectal cancer. After 50 days, the nude mice were killed and the liver metastases were observed at the gross level and under microscope. Results: the liver metastasis model of colorectal cancer in nude mice was successfully constructed and confirmed by H-E staining. 鍚勭粏鑳炲共鎵扮粍(SW480-KD,HT29-KD)瑁搁紶浣撹川閲忔槑鏄句綆浜庣浉搴旂┖鐧藉�圭収缁勫拰闃存�у�圭収缁刐SW480-KD:(15.02卤2.00)vs(18.00卤2.48),(18.03卤2.05)g;F=3.761,P0.05;HT29-KD:(15.39卤2.16)vs(17.96卤2.48),(18.30卤1.87)g;F=3.721,P0.05].SW480-KD缁勮倽杞�绉荤槫鏁扮洰鏄庢樉澶氫簬SW480-CON缁勫拰SW480-NC缁刐(57.83卤22.56)vs(29.50卤16.90),(28.20卤15.40)涓，

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