向少龙,赵艳,刘娟娟,赵厚育
XIANG Shaolong,ZHAO Yan,LIU Juanjuan,ZHAO Houyu

• 1:贵州医科大学
• 2:贵州医科大学分子生物学重点实验室
• 3:贵州医科大学附院耳鼻咽喉科

摘要(Abstract)：

目的:探讨沉默缺氧诱导因子-1α对人鼻咽癌裸鼠移植瘤生长的影响。方法:15只4~6周龄雌性裸鼠,随机均分为为HIF-1α干扰组、空载组及阴性对照组;采用脂质体转染法将HIF-1α的microRNA干扰质粒转染CNE-2细胞,RT-q PCR检测其沉默效应,将转染HIF-1α干扰质粒的CNE-2细胞、转染空载质粒的CNE-2细胞以及CNE-2细胞分别接种于3组裸鼠右腋皮下,建立鼻咽癌CNE-2细胞裸鼠移植瘤模型,形成肉眼可见的肿瘤后再喂养16 d,观察各组成瘤时间及肿瘤体积;RT-q PCR检测3组移植瘤中HIF-1α基因的表达,蛋白质印迹法检测3组移植瘤中HIF-1α蛋白的表达。结果:成功构建裸鼠移植瘤模型,HIF-1α干扰组、阴性对照组、空载组裸鼠成瘤时间分别为(8.60±1.40)、(4.60±0.55)、(4.80±0.84)d,差异有统计学意义(P<0.05);肿瘤体积分别为(128.60±77.70)、(1 411.10±212.50)、(1 306.80±154.60)mm3,差异有统计学意义(P<0.05);HIF-1α干扰组瘤体组织中的HIF-1α基因和蛋白的表达量明显降低(P<0.05)。结论:沉默HIF-1α基因能有效抑制人鼻咽癌裸鼠移植瘤成瘤时间和肿瘤的生长。
Objective: To investigate the influence of silencing hypoxia inducible factor 1-alpha( HIF-1α) on growth of human nasopharyngeal carcinoma xenografts in nude mice. Methods: Respectively15 female nude mice aged 4 to 6 weeks were randomly and equally divided into HIF-1α interference group( group H),vector group( group V),and negative control group( group C). CNE-2 cells were transfected with microRNA interference plasmids of HIF-1α by using lipofection. RT-q PCR was used to detect the silencing effect. CNE-2 cells that transfected by HIF-1α interference plasmids,that by blank plasmids,and that non-transfected were used to inoculate subcutaneously into the right underarm of the nude mice in 3 groups respectively to establish xenografts model. In 16 days after the cacinoma appeared,the tumor forming time was observed,and tumor volume was measured and calculated. The mRNA and protein expression levels of HIF-1α in xenografted tumors were detected by using RT-q PCR and Western blotting. Results: the nasopharyngeal carcinoma xenografts nude mice models were successfully established. The macroscopic tumor forming time in groups H,C,and V were( 8. 6 ± 1. 4)d,( 4. 6 ± 0. 55) d,and( 4. 8 ± 0. 84) d respectively,and the differences were statistically significant( P < 0. 05). The final tumor volumes of these groups were( 128. 6 ± 77. 7) mm3,( 1411. 1 ± 212. 5)mm3,and( 1306. 8 ± 154. 6) mm3 respectively. The mRNA and protein expression levels of group H were significantly lower than those of groups V and C( P < 0. 05). Conclusions: silencing HIF-1αgene can effectively inhibit the growth of human nasopharyngeal carcinoma xenografts in nude mice.

关键词(KeyWords)： 鼻咽肿瘤;缺氧诱导因子-1α;肿瘤移植;RNA干扰
nasopharyngeal neoplasms;hypoxia inducible factor 1-alpha;neoplasm transplantation;RNA interference

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金地区科学基金项目(81260353)

作者(Author): 向少龙,赵艳,刘娟娟,赵厚育
XIANG Shaolong,ZHAO Yan,LIU Juanjuan,ZHAO Houyu

DOI: 10.19367/j.cnki.1000-2707.2017.04.005

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