摘要
目的研究外源性突变型p27基因(p27mt)对裸鼠皮下肝癌移植瘤侵袭性生长的影响。方法应用细胞移植法把人原发性肝癌细胞株SMMC-7721种于BALB/c裸鼠皮下,构建人肝癌裸鼠皮下移植瘤动物模型,将PBS液(100μl)、腺病毒介导的突变型p27(Ad-p27mt,5.0×109pfu,100μl)及Lac-Z(Ad-Lac-Z,5.0×109pfu,100μl)分别注射到瘤体后,每隔3天测量肿瘤体积,计算肿瘤生长抑制率。于处理后21天取材行冰冻切片,免疫组织荧光化学法观察各组移植瘤的P27蛋白、CD44v6及Ⅷ因子的表达,数码成像后,用Image-ProPlus软件计算荧光亮度和细胞总数,测量积分吸光度(integrated optical density,IOD),计算平均吸光度(Average optical density,AOD),以平均吸光度来反映细胞因子表达水平。结果与Ad-Lac-Z、空白对照组比较,Ad-p27组移植瘤P27mt蛋白表达明显增高(P<0.01),并且过表达的P27mt蛋白明显减小了移植瘤体积,抑制了肿瘤的生长速度;血管大体观发现Ad-p27组血管不仅细而且分支少,Ⅷ因子免疫组织荧光化学观察发现瘤体内血管密度明显降低;同时Ad-p27组也明显降低了肿瘤细胞CD44v6(P<0.05)。结论 Ad-p27mt能在裸鼠SMMC-7721肝癌皮下移植瘤过表达P27mt蛋白,不仅通过抑制血管的生成而抑制肿瘤的生长,而且通过降低肿瘤细胞CD44v6的表达而显著抑制移植瘤的侵袭性。
Objective To study the effects of mutant p27（p27 mt） gene on invasive growth of SMMC-7721（HCC） in nude mice.Methods A mouse model of human hepatocellular carcinoma（SMMC-7721） was established by subcutaneous transplantation in BALB/c mice.The PBS（100μl）,adenovirus-mediated mutant p27（Ad-p27mt,5.0×109 pfu,100μl） and Lac-Z（Ad-Lac-Z,5.0×109 pfu,100μl） were injected into the transplanted tumors.Subsequently,the tumor size was tested in every 3 days.The expression of p27 protein,CD44v6 and Ⅷ factor in the transplanted tumors were detected through immunofluorescence method and Semi-quantitative analysis of the expression of P27 protein,CD44v6 and Ⅷ factor were evaluated by integral optical density（IOD） and average optical density（AOD） through Image-Pro Plus software.Results Compared with the Ad-Lac-Z and blank control group,P27 protein expression in Ad-p27 group was obviously increased（P0.01）,and the expression of CD44v6 and Ⅷ factor were significantly decreased（P0.05）.Conclusion Over-expressing p27 mt could significantly inhibit the growth of SMMC-7721 in nude mice through angiogenesis inhibition,and obviously decreased the invasive ability of SMMC-7721 through CD44v6 expression down-regulation.
出处
《肿瘤防治研究》
CAS
CSCD
北大核心
2010年第10期1123-1127,共5页
Cancer Research on Prevention and Treatment
基金
湖北省自然科学基金项目(2005ABA082)
关键词
p27mt
肝癌
裸鼠
生长
侵袭
p27mt
Hepatocellular carcinoma
Nude mice
Growth
Invasion
