Jab1基因沉默对裸鼠人喉癌移植瘤生长影响的研究  

Inhibitory effect of Jabl-specific shRNA on the progression of human laryngeal carcinoma xenografts nude mice

作　　者：李海霞[1] 曹华[1] 

机构地区：[1]郑州大学第一附属医院耳鼻咽喉科,450052

出　　处：《中华耳鼻咽喉头颈外科杂志》2012年第5期406-410,共5页Chinese Journal of Otorhinolaryngology Head and Neck Surgery

摘　　要：目的探讨Jab1重组质粒对人喉鳞状细胞癌（简称鳞癌）Hep．2细胞裸鼠移植瘤生长的影响。方法利用喉鳞癌Hep-2细胞裸鼠皮下移植瘤模型，通过向瘤体内注射pJab1、阴性对照质粒和生理盐水，观察瘤体增长情况，通过免疫组化方法、反转录聚合酶链反应（RT-PCR）观察瘤体内Jab1、p27的mRNA及蛋白表达水平的变化。结果pJab1质粒组的肿瘤体积为（267．60±88．19）mm3（i±s，以下同），与阴性对照组的（832．20±140．39）mm3及生理盐水组的（895．40±145．93）mm3相比，差异有统计学意义（F=36．73，P〈0．001）；免疫组化结果显示pJab1质粒组瘤内的Jabl蛋白的表达率降低为32．40％±5．59％，p27蛋白的表达率增高到76．80％±6．30％（P〈0．001），与阴性对照组及生理盐水组比较，差异有统计学意义；RT—PCR结果显示实验组瘤内Jab1的mRNA相对表达水平降低至0．65±0．03（F=558．00，P〈0．001），p27的mRNA无明显变化，为0．80±0．02（F=1．52，P〉0．05）。结论pJab1干扰质粒可明显降低裸鼠肿瘤组织内Jab1基因的表达并抑制瘤体的生长；pJab1质粒能有效抑制Jab1基因的表达，有望成为临床治疗喉癌的新途径。Objective To evaluate the antitumour efficacy of shRNA plasmid specifically targeting Jabl gene. Methods The nude mouse tumor model was made by subcutaneous injection of human laryngeal carcinoma Hep-2 cells. The tumor growth was monitored after intratumoral injection of pJabl, pKB plasmids and saline. Jabl and p27 expressions in turnout tissues were examined by immunohistoehemistry stainingand RT-PCR. Results Mean volume of the pJabl-treated tomors was（267.60 ± 88.19）mm3, significantly less than that of tumors treated with pKB plasmids （ 832.20 ± 140.39 ） mm3 or saline （ 895.40 ± 145.93 ） mm3 （ F = 36.73, P 〈 0.001 ）. Immunohistochemistry showed that the expression of Jabl protein was significantly reduced in the pJabl-treated group （ 32. 40% ± 5.59% ） compared to the control groups, whereas the expression rate of p27 protein in the pJabl group（76.80% ± 6.30% ） was significantly increased compared to the control groups（P 〈0.001 ）. The down regulation of Jabl protein by pJabl plasmid was consistent with mRNA expression confirmed by RT-PCR. The level of Jabl mRNA level in the pJabl-treated group（0.65 ± 0.03 ） was significantly lower than the control groups （ F = 558.00, P 〈 0.001 ）, however, p27 mRNA, was 0.80 ± 0.02, had no significant alteration（ F = 1.52, P 〉 0.05 ）. Conclusions The pJabl plasmid results in downregulation of Jabl in an xenograft tumour model of human laryngeal carcinoma Hep-2 cells and significantly inhibits the turnout growth in vivo. This suggests that pJabl plasmid specifically targeting Jabl gene expression could be an effective therapy for human laryngeal carcinoma.

关 键 词：喉肿瘤 癌 鳞状细胞 肽水解酶类 细胞内信号肽和蛋白质类 基因疗法 模型 动物 

分 类 号：R739.65[医药卫生—肿瘤] R-332[医药卫生—临床医学]