摘要
背景与目的:肝转移是晚期结肠癌患者最常见的致死原因,也是最常见的内脏转移,高达50%以上的结肠癌患者都会出现肝转移。本研究探讨arresten基因转染对人结肠癌LoVo细胞形成的裸鼠实验性结肠癌肝转移的影响。方法:通过脂质体转染法将arresten基因导入LoVo细胞,RT-PCR、Western blot分别检测arresten在mRNA、蛋白水平的表达,四甲基噻唑蓝(MTT)比色法检测arresten对LoVo细胞增殖的影响;通过建立裸鼠实验性结肠癌肝转移模型了解arresten对肿瘤转移的抑制作用;FⅧRag多克隆抗体染色的免疫组化方法检测肿瘤组织的微血管密度(microvessel density,MVD)。结果:RT-PCR、Westernblot结果显示arresten基因成功导入LoVo细胞并有arresten蛋白的表达。MTT比色法显示不同浓度的arresten对LoVo细胞增殖的影响差异无统计学意义(P>0.05)。导入arresten基因的LoVo细胞转移率为(25.1±2.1)%,低于未导入arresten基因的LoVo细胞的(87.1±1.2)%和对照组的(87.1±1.5)%,其差异均有统计学意义(P值均<0.05)。pSecTag2-arresten组裸鼠形成的肿瘤结节数为4.5±0.5,低于另外两组的19.6±2.5和20.4±2.5,其差异均有统计学意义(P值均<0.05)。pSecTag2-arresten组形成的肿瘤MVD为15.3±3.5,低于另外两组(分别为42.2±2.6、45.6±5.1),其差异均有统计学意义(P值均<0.05)。结论:arresten能抑制结肠癌肝转移,其作用机制可能与arresten抑制肿瘤血管生成有关。
BACKGROUND ＆ OBJECTIVE= Liver metastasis is the most common cause of death due to colorectal cancer. Above 50% colorecal cancer patients have liver metastasis. This study was to investigate the effect of transfection of arresten gene on liver metastasis from human colorectal cancer （LoVo） xenografts in nude mice. METHODS. The eukaryotic expression plasmid pSecTag2-arresten was transfected into human colorectal cancer cell line LoVo using Lipofectamine 2000. Cells were divided into pSecTag2-arresten group, pSecTag2 group and control group. Expressions of arresten at mRNA and protein levels were detected by RT-PCR and Western blot, respectively. The effect of arresten on proliferation of LoVo cells was measured using MTT assay. LoVo cells transfected with pSecTag2- arresten were implanted into nude mice to investigate the effect of arresten on hepatic metastasis of colorectal cancer. Microvessel density （MVD） of xenograft tumors was assessed using immunohistochemistry with FVIIIRag monoclonal antibody. RESULTS= Arresten was successfully transfected and expressed in LoVo cells. Inhibition of cell proliferation did not differ significantly in all three groups （P 〉0.05）. The metastastic rate was lower in pSecTag2-arresten group [ （25.1 ±2.1）% ] than in pSecTag2 group [ （87,1 ± 1.2）% or control group [（87.1±1.5）%] in LoVo cells （P〈0.05）. The number of xenograft tumors and MVD were higher in pSecTag2-arresten group [（4.5 ±0.5） and （15.3±3.5）J than in pSecTag2 group [（19.6±2.5） and （42.2± 2.6）1 or in control group E（20.4±2.5）and （45.6±5.1）1 in nude mice. CONCLUSION. Arresten can inhibit hepatic metastasis from colorectal cancer, which may be through its inhibition on tumor angiogenesis.
出处
《癌症》
SCIE
CAS
CSCD
北大核心
2008年第10期1039-1043,共5页
Chinese Journal of Cancer
基金
国家青年科学基金(编号:30700803)~~
