摘要
目的 观察重组人内皮抑素腺病毒(Ad/hEndo)对人肝癌裸鼠移植瘤生长的影响。方法 人脐静脉内皮细胞ECV-304经Ad/hEndo感染后,western印迹检测人内皮抑素的表达。人肝癌BEL-7402细胞移植到裸鼠背脊部后,检测Ad/hEndo对肝癌移植瘤生长的抑制作用。逆转录聚合酶链反应(RT-PCR)检测肿瘤组织中内皮抑素mRNA的表达。分析人内皮抑素在裸鼠体内的表达分布。结果 Western印迹检测到人内皮抑素基因在ECV-304细胞内高效表达。Ad/hEndo明显抑制人肝癌BEL-7402裸鼠移植瘤生长(F=4.061,P<0.05)。Ad/hEndo组血管密度计数为6.88±1.08,DMEM组为13.60±1.71(t=9.216,P<0.01)。瘤内注射Ad/hEndo后3d,RT-PCR在肿瘤组织检测到内皮抑素mRNA的表达,7d后表达不明显。人内皮抑素蛋白主要分布在肿瘤组织。结论 腺病毒介导的人内皮抑素基因在体内、体外获得高效表达,并明显抑制肝癌裸鼠移植瘤的生长与血管生成。
Objective To investigate the inhibitive effect on the growth of hepatocellular carcinoma (HCC) xenografted in nude mice by adenovirus-mediated human endostatin gene. Methods The expression efficiency of endostatin was examined after ECV-304 cells infected with Ad/hEndo by western blot. The hepatoma BEL-7402 cells were injected into Balb/c nude mice to detect the inhibition of Ad/hEndo on the growth of HCC xenografted in nude mice. The expression of endostatin mRNA in tumor tissue was analyzed with RT-PCR, and its distribution in vivo was also analyzed. Results High level expression of endostatin achieved in infected ECV-304 cells by western blot. Ad/hEndo significantly inhibited the growth of xenografted BEL-7402 tumors (F = 4.061, P < 0.05). The intratumoral microvessel density (MVD) decreased significantly in the treated mice (6.88±1.08 vs 13.60±1.71, t = 9.216, P < 0.01). The expression of endostatin mRNA in tumor tissue was detected by RT-PCR in 3 days after administration intratumorally with Ad/hEndo and almost disappeared in 7 days. Endostatin mRNA was mainly located in tumor tissue with a higher concentration than that in heart, lung, spleen and liver after Ad/hEndo administration. Conclusion Adenovirus-mediated human endostatin gene can be expressed efficiently in vitro and in vivo, and significantly inhibit the growth of BEL-7402 xenografted tumors in nude mice.
出处
《中华肝脏病杂志》
CAS
CSCD
2003年第9期542-545,共4页
Chinese Journal of Hepatology
基金
国家高技术研究发展863计划(2001AA217081)
