anti-Livin核苷酸调低喉鳞癌裸鼠移植瘤内Livin的表达对喉鳞癌细胞凋亡的影响

摘要

目的 利用anti-Livin核苷酸调低喉鳞癌裸鼠移植瘤内Livin的表达量,探讨其对喉鳞癌细胞凋亡的影响. 方法28只雌性裸鼠(BALB/c-nu/nu)随机分为Livin AS-DNs(Livin反义多核苷酸)、Mis-DNs、Sen-DNs和对照组,每组7只,于各组裸鼠背部皮下注射1×105Hep-2细胞成瘤,用AS-DNs、Mis-DNs、Sen-DNs和生理盐水分别对裸鼠移植瘤进行注射;测量肿瘤长径和短径变化,将肿瘤体积变化绘制成生长曲线.采用Western blot检测AS-DNs、Mis-DNs组、Sen-DNs组、对照组瘤组织Livin蛋白;利用TUNEL染色法检测各组瘤体中细胞凋亡. 结果 Livin蛋白表达最低为AS-DNs组(15.02 ± 1.68)%,Mis-DNs组、Sen-DNs组和对照组明显高于AS-DNs组(P<0.05);AS-DNs组裸鼠瘤体内喉鳞癌细胞的凋亡指数最高为(20.15 ± 3.1)%,明显高于Mis-DNs组、Sen-DNs组和对照组(P<0.05);分别对裸鼠移植瘤进行注射后,AS-DNs治疗组的喉鳞癌细胞生长较Mis-DNs组、Sen-DNs组和对照组慢,差异有统计学意义(P<0.05). 结论 反义Livin多核苷酸在裸鼠移植瘤内促进了喉鳞状细胞癌的凋亡,抑制了肿瘤的生长.%Objective To explore the effect of antisense oligodeoxynucleotides targeting Livin on cell apoptosis of laryngeal squamous cell carcinoma in transplantation tumor of nude mice. Methods Female BALB/c-nu/nu mice were divided into four groups(n=7 in each group):control group,AS-DNs group,Sen-DNs group,and Mis-DNs group. The 1×105Hep-2 cells were subcutaneously in-oculated in dorsal space of the mice to generate the transplanted tumor,and the xenograft was treated with AS-DNs,Mis-DNs,Sen-DNs and normal saline in four groups,respectively. The major axis and minor axis of xenograft were determined,and then the growth curve of tumor volume was drawn. The expression of Livin protein in tumor was examined by Western blotting. TUNEL assay was used to analyze Hep-2 cell apoptosis in tumor of nude mice. Results The Livin protein expression was(15.02 ± 1.68)% in AS-DNs group,and lower than that in the other three groups(P<0.05).The apoptotic index of Hep-2 cells was(20.15 ± 3.1)% in AS-DNs group,which was significantly higher than that in the other groups(P<0.05). The tumor growth rate was slower in AS-DNs group than that of control group,Sen-DNs group,and Mis-DNs group(P<0.05). Conclusion AS-DNs of Livin could promote the apoptosis of laryngeal squamous cell carcinoma cell and inhibit the tumor growth in vivo.