p27mt对裸鼠肝癌移植瘤侵袭性生长的影响

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, 5.0×109 pfu, 100μl)及Lac Z(Ad Lac Z, 5.0×109 pfu, 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-Pro Plus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。

Effects of Mutant p27 Gene on Invasive Growth of Hepatocellular Carcinoma in Nude Mice

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 (P<0.01), and the expression of CD44v6 and Ⅷ factor were significantly decreased (P<0.05). 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.