摘要
目的探讨重组慢病毒介导的基质金属蛋白酶-2（MMP-2）和基质金属蛋白酶-9（MMP-9）基因同时沉默对喉鳞癌侵袭生长的抑制作用。方法采用核糖核酸（RNA）干扰技术，运用MMP-2和MMP-9基因小干扰RNA（siRNA）的重组慢病毒（MMP-2-RNAi-lentivirus，MMP-9-RNAi-lentivirus）共同转导喉鳞癌Hep-2细胞，并运用逆转录聚合酶链反应方法（RT—PCR）检测Hep-2细胞MMP-2和MMP-9基因的表达。通过Boyden小室实验观察MMP-2-RNAi-lentivirus和MMP-9-RNAi-lentivirus重组慢病毒转导对喉癌细胞侵袭能力的影响。构建喉癌移植瘤模型，将MMP-2-RNAi-lentivirus和MMP-9-RNAi-lentivirus重组慢病毒瘤内注射，观察抑瘤效果。运用免疫组化方法检测移植瘤中增殖细胞核抗原（PCNA）的表达，评估其对细胞增殖的影响。结果重组慢病毒MMP-2-RNAi—lentivirus和MMP-9-RNAi—lentivirus能高效地转导入靶细胞。RT-PCR检测显示，转导后的Hep-2细胞中MMP-2和MMP-9表达阴性。侵袭实验显示，实验组Hep-2细胞中穿过人工基底膜的细胞数为（14±4）个，明显少于空载体对照组的（32±6）个（P〈0．01）。体内实验显示，MMP-2-RNAi-lentivirus和MMPO-RNAi-lentivirus重组慢病毒瘤内注射后，裸鼠移植瘤平均重量和平均体积明显小于对照组，抑瘤率为46．59％，PCNA指数也明显降低。结论慢病毒介导的MMP-2和MMP-9基因共同沉默能有效的抑制喉癌的侵袭，生长及增殖。
Objective To investigate the inhibitive role of the invasion and growth of laryngeal cancer by lentivirus mediated Matrix Metalloproteinase-2 （MMP-2） and Matrix Metalloproteinase-2 （MMP-9） gene silence. Methods RNA interference （RNAi） technic was used. Recombinant lentivirus of small interference RNA targeting MMP-2 and MMP-9 gene were transduced into Hep-2 cells, and Reverse transcriptase polymerase chain reaction （RT-PCR） was used to test MMP-2 and MMP-9 expression in Hep-2 cells. The effect of the invasive capability in Hep-2 cells after MMP-2-RNAi-lentivirus and MMP-9-RNAi- lentivirus transduction was observed by Boyden assay. In vivo experiment, the nude mouse model of laryngeal squamous carcinoma was established and MMP-2-RNAi-lentivirus and MMP-9-RNAi-lentivirus were intratumoral injected. Finally, the tumor inhibitive effect was observed and the Proliferating cell nuclear antigen （PCNA） expression in xenografts were examined to evaluate proliferation change of the Hep-2 cells. Results The recombinant lentivirus can transduce the target cells efficiently. RT-PCR showed the expression of MMP-2 and MMP-9 were negative. Boyden assay showed there were （ 14 ± 4） Hep-2 cells permeate artificial basement which was less than that （32 ±6 ） in the control group. In vivo, the average tumor weight and volume in treated group were significantly lower than those in the control group （ P 〈 0. 01 ） and the tumor inhibitive rate was 46.59%. Immunohistochemistry showed PCNA index in treated group was obviously lower than that in control group （P 〈 0. 01 ）. Conclusion The invasion, growth and proliferation of laryngeal cancer can be inhibited by lentivirus mediated MMP-2 and MMP-9 gene silence together.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2008年第1期36-39,共4页
National Medical Journal of China
基金
国家自然科学基金资助项目（30540010）
中国博士后科学基金资助项目（20070410920）
黑龙江省青年基金资助项目（QC06C054）
黑龙江省海外学人科研基金资助项目（1151hz029）.
关键词
慢病毒属
喉肿瘤
基质金属蛋白酶
Lentivirus
Laryngeal neoplasms
Matrix metaUoproteinases
