CXCR4小干扰RNA对人喉鳞状细胞癌Hep-2细胞裸鼠移植瘤生长的抑制作用  

Inhibitory effect of CXCR4-Targeted small interfering RNA mediated by liposome on mice model of laryngeal carcinoma

作　　者：唐凤翔[1] 胡国华[2] 

机构地区：[1]重庆市第九人民医院耳鼻咽喉科,重庆400700 [2]重庆医科大学附属第一医院耳鼻咽喉科,重庆400016

出　　处：《中国耳鼻咽喉头颈外科》2011年第10期525-528,共4页Chinese Archives of Otolaryngology-Head and Neck Surgery

摘　　要：目的探讨CXC族趋化因子受体4(CXCchemokine receptor 4,CXCR4)小干扰RNA(smallinterfering RNA,siRNA)对人喉癌细胞株Hep-2细胞裸鼠体形成的移植瘤淋巴结微转移的影响。方法将与CXCR4mRNA互补的siRNA通过阳离子脂质体转染Hep-2细胞,用RT-PCR方法检测转染后Hep-2细胞的CXCR4 mRNA表达情况。建立人喉癌裸鼠皮下移植瘤模型,绘制裸鼠生长曲线,计算肿瘤抑制率。处死裸鼠后测量移植瘤体积并计数淋巴结转移情况,HE染色病理证实移植瘤及淋巴结转移,免疫组化SP法检测各组皮下移植瘤CXCR4蛋白的表达情况。结果人喉癌裸鼠皮下移植瘤模型建立成功后,siRNA转染组裸鼠皮下移植瘤的体积增长明显滞后于Hep-2细胞组,瘤体质量下降;HE染色显示Hep-2组肿瘤体积大、异形明显。免疫组化结果显示siRNA转染组CXCR4蛋白表达明细减少。结论 CXCR4 siRNA能在体内明显抑制喉癌CXCR4基因的表达,进而抑制喉癌实体瘤增殖和生长,且能在体内有效下调CXCR4蛋白的表达。OBJECTIVE To investigate the effect of CXCR4-targeted small interfering RNA（siRNA） mediated by liposome vector in inhibiting the growth of human laryngeal carcinoma cell Hep-2 xenografts in nude mice.METHODS The siRNA targeting CXCR4 mRNA was transfected into Hep-2 cells via liposome Lipofectin.Semi-quantitative RT-PCR was performed to evaluated the levels of CXCR4 gene expression.Nude mice models of laryngeal carcinoma were estabilished.The growth curves of the mice were drawed.And tumor control rates were calculated.After the mice were sacrificed,the volume of the transplanted tumor was evaluated.And the number of the metastatic lymph nodes was counted.Using hematoxylin-eosin（HE）staining,the transplanted tumor and metastatic lymph node were confirmed.CXCR4 protein expression of tumors was investigated by immunohistochemistry.RESULTS Compared to controls,the enlargement of tumor volume significantly lagged in transfected group of siRNA-CXCR4.And the mass of the tumor was decreased in transfected group of siRNACXCR4.HE staining showed the hep-2 cells of the tumor were bigger and obviously heteromorphic.The expression of CXCR4 mRNA and protein in the siRNA transfected tumor was obviously reduced.CONCLUSION CXCR4 siRNA targeting CXCR4 mRNA can markedly inhibit the growth of laryngeal carcinoma cell xenografts in nude mice through lastingly inhibition effect on CXCR4.Also it can down regulate the expression of CXCR4 protein in vivo.

关 键 词：趋化因子 CXC 癌 鳞状细胞 RNA 小分子干扰 转染 免疫组织化学 

分 类 号：R739.65[医药卫生—肿瘤]