RNAi沉默Livin基因对裸鼠移植瘤生长及Ki67蛋白表达的影响

RNAi silence Livin gene in nude mice transplantation tumor growth and influence of Ki67 protein expression

目的观察RNAi沉默Livin基因在裸鼠移植瘤生长及Ki67蛋白表达中的影响。方法 BALB/c裸鼠30只,在裸鼠右侧腋下构建荷瘤模型,待肿瘤生长至100 mm3左右,建立裸鼠皮下移植瘤模型。将裸鼠随机分为三组,A组(空白对照)、B组(阴性载体对照组)及C组(实验组)。A组为空白对照,瘤内注射0.9%氯化钠溶液5μl,注射时注意四周加中央5点进行注射;B组裸鼠瘤内注射携带无关序列的慢病毒颗粒5μl,B、C组注射方式均与A组一致;C组裸鼠瘤内注射Livin基因shRNA慢病毒颗粒5μl。采用免疫组织化学的方法检测Livin蛋白和Ki67蛋白。结果 C组瘤体生长较缓慢,瘤体质量明显低于A组和B组,而A组和B组两组瘤体质量差异无统计学意义(P>0.05);三组Livin蛋白阳性细胞着色以棕黄色为主,A组、B组Livin蛋白阳性细胞百分率(55.41%±12.40%比55.43%±8.12%)比较,差异无显著性(P>0.05),C组(20.54%±7.53%)较前两组则明显降低,Livin蛋白Berry N评分C组明显低于其他两组;A组和B组Ki67蛋白阳性表达的细胞核着色强度为棕褐色,C组着色不均匀,颜色明显变浅,三组Ki67蛋白阳性细胞百分率比较,A组(95.46%±3.41%)和B组(94.12%±4.10%)比较,差异无统计学意义(P>0.05),C组(68.41%±11.42%)则明显减少,Ki67蛋白Berry N评分C组明显低于其他两组(P<0.05)。Livin蛋白与Ki67蛋白阳性细胞百分比呈正相关(r=0.63,P<0.05)。结论 RNAi沉默Livin基因可抑制移植瘤生长,降低Livin蛋白、Ki67蛋白的表达。Livin蛋白、Ki67蛋白两者呈正相关。

Objective To observe the RNAi silence Livin gene in nude mice transplantation tumor growth and influence of Ki67 protein expression. Methods Right axillary tumor - burdened model was constructed in 30 BALB/c nude mice, in nude mice with tumor growth to about 100 mm^3. The nude mice subcutaneous transplantation tumor model is established. Nude mice were randomly divided into three groups, group A （ck） and carrier （negative control group） and group B （experimental group）, group C for blank control group. 5 μl tumor injection of 0.9% so- dium chloride solution, injection around when you add the 5 points. Group B in the nude mice tumor injection with slow virus particles is sequen- tial 5 μl. The injection mode B and C group were consistent with the group A. Group C nude mice tumor injection Livin shRNA gene slow virus particles within 5 μl. The method was used to detect Livin protein and Ki67 protein. Results Group C tumors grew more slowly. The quality of tumors was lower than that in group A and group B. The quality tumors of Group A and group B had no statistically signifi- cant differences in the two groups（ P 〉0.05 ）. Three groups of Livin protein positive cells color is given priority to with tan. Group A and group B Livin protein positive ceils percentage were compared （55.41% ± 12.40% vs. 55.43% ± 8.12% ）, there was no significant difference（ P 〉 0.05 ）, compared with the previous two group C group is significantly lower. The level of group C is about （ 20.54%±7.53% ）. Livin protein Berry N score group C was lower than that in other two groups. Group A and group B Ki67 protein positive expression of nucleus tinting strength for tan. The group C showed uneven color. The color of group C becomes shallow. The Ki67 protein positive cells percentage of three groups were compared, group A （95.46%±3.41% ） and group B （94.12%±4.10% ）. There was no statistical difference （ P 〉0.05）. Group C is signifi- cantly reduced, which was about the （68.41%± 11.42% ）. Berry Ki67 protein N score group C was lower than that in other two groups （ P 〈 0.05 ）. Livin protein percentage and Ki67 protein positive cells were positively correlated （ r = 0.63, P 〈 0.05 ）. Conclusion RNAi silence Livin gene can inhibit the growth of transplanted tumor, reduce the expression of Livin protein and Ki67 protein. Livin protein and Ki67 protein was positively related to both.