全长型脾酪氨酸激酶基因转染对人喉鳞癌裸鼠移植瘤的生长抑制作用  

Inhibitory Effect of SYK( L) Gene Therapy on Human Laryngeal Carcinoma Xenograft Model in Nude Mice by Cationic Liposome Mediated Transfection

作　　者：张朝晖[1] 李志海[1] 蔡志毅[1] 陶宝鸿[1] 金巧智[1] 

机构地区：[1]台州学院医学院附属市立医院耳鼻咽喉科,318000

出　　处：《医学研究杂志》2015年第6期65-69,共5页Journal of Medical Research

基　　金：浙江省中医药科学研究基金资助项目(2013ZA132);台州市科技计划项目(2009KYB241)

摘　　要：目的探讨全长型脾酪氨酸激酶[full-length spleen tylosine kinase,SYK(L)]基因转染对人喉鳞状细胞癌Hep-2细胞裸鼠移植瘤生长的影响。方法建立人喉鳞癌Hep-2细胞裸鼠皮下移植瘤模型,分为SYK(L)重组质粒转染组、阴性对照质粒转染组和生理盐水组,分别向瘤体内注射SYK(L)重组质粒p IRES2-EGFP-SYK(L)及脂质体混合物、阴性对照质粒p IRES2-EGFP及脂质体混合物和生理盐水,观察肿瘤生长情况。通过实时荧光定量聚合酶链式反应(quantitative real time fluorescence polymerase chain reaction,Q-RT-PCR)、Western blot法检测各组瘤体内的SYK(L)mRNA及蛋白表达水平。结果 SYK(L)重组质粒转染组的肿瘤平均体积为367.61±81.23mm3,明显小于阴性对照质粒转染组900.20±131.41mm3及生理盐水组930.71±143.73mm3,差异有统计学意义(F=99.91,P<0.01);Q-RT-PCR结果显示SYK(L)重组质粒转染组瘤体内mRNA相对表达量为2.268±0.075,明显高于阴性对照质粒转染组(1.212±0.025)及生理盐水组(1.175±0.031)(F=102.01,P<0.01);Western blot法检测结果显示SYK(L)重组质粒转染组瘤体内相对蛋白含量(0.834±0.021)明显高于阴性对照质粒转染组(0.243±0.041)和生理盐水组(0.301±0.039),差异有统计学意义(F=104.13,P<0.01)。结论全长型脾酪氨酸激酶基因转染对人喉鳞癌裸鼠移植瘤有明显的生长抑制作用,有望成为基因治疗喉癌的新靶点及途径。Objective To observe the effect of cationic liposome mediated full - length spleen tylosine kinase [ SYK（L） ] gene trans- fection on the growth of human laryngeal cancer Hep -2 cells in the nude mice. Methods 15 nude mouse tumor models were made by subcutaneous injection of human laryngeal carcinoma Hep - 2 cell. Then they were randomly and equally divided into 3 groups. The tumor growth was monitored after intratumoral injection of SYK（L） overexpression vector plRES2 -EGFP- SYK（L） ,plasmid vector pIRES2 - EGFP and saline. The gene and protein expression of SYK（L） expressions in tumour tissues were detected by Q -RT- PCR and wetern blot. Results Mean volume of the SYK （L） overexpression vector plRES2 - EGFP - SYK （L） - treated tumors was 367. 61 ± 81.23mm3 , significantly less than that of tumors treated with plasmid vector plRES2 - EGFP 900.20± 131.41mm3 or saline 930.71 ± 143.73 mm3 （ F = 99.91, P 〈 0.01 ）. Q - RT - PCR analysis indicated that the 2 - aaCT of SYK （ L ） mRNA in SYK （L） overexpression vector pIRES2 -EGFP- SyK（L） -treated group（2. 268 ± 0. 075）was significantly higher than that in plasmid vector pIRES2 -EGFP- treated group （1. 212 ± 0. 025 ）and saline treated group （1. 175 ± 0. 031 ） （F = 102.01,P 〈 0.01 ）. Western blot showed that the level of SYK （L） protein in SYK（L） overexpression vector pIRES2 - EGFP - SYK（L） - treated group （0. 834 ± 0. 021 ） was significantly higher than that in plasmid vector pIRES2 - EGFP - treated group （0. 243 ± 0. 041 ） and saline treated group （0. 301 ± 0. 039 ） （ F = 104.13, P 〈 0.01 ）. Conclusion The growth of human laryngeal carcinoma xenograft in nude mice could be inhibited significantly by SYK（L） gene transfection. This suggests that SYK（L） gene transfection could be an effective therapy for human laryngeal carcinoma.

关 键 词：全长型脾酪氨酸激酶 喉癌 移植瘤 抑制 基因疗法 

分 类 号：R739.6[医药卫生—肿瘤]