摘要
目的:探讨Bcl-2基因的小干扰RNA(siRNA)真核表达载体干扰Bcl-2基因表达对人胆管癌细胞QBC939、胆囊癌细胞GBC-SD裸鼠体内移植瘤生长的抑制作用。方法:分别制作人胆管癌细胞QBC939、胆囊癌细胞GBC-SD细胞株移植瘤模型,随机分为3组,pSilencerTM-EGFP sh515组(实验组)、pSilencerTM-EGFP shCon组(空载体阴性对照组)和对照组。实验组用Bcl-2基因的小干扰RNA(siRNA)真核表达载体多点注射于移植瘤周围,空载体阴性对照组用空载体多点注射于移植瘤周围,对照组不作处理,观察其生长情况,计算肿瘤生长体积、生长率。6周后,处死裸鼠,剥离瘤体组织进行称重,并采用免疫组化检测Bcl-2蛋白的表达。结果:成功建立裸鼠人胆管癌细胞QBC939,胆囊癌GBC-SD细胞的动物模型。胆管癌细胞实验组、空载体阴性对照组、对照组瘤体平均体积(mm3)分别为:801.776±59.6,1383.980±78.2,1490.235±89.0。瘤体平均瘤体重量(g)为:0.90±0.11,1.49±0.31,1.63±0.22。平均生长率为:20.558%,35.587%,38.211%。人胆囊癌细胞实验组、空载体阴性对照组、对照组瘤体平均体积(mm3)分别为:729.736±66.6,1493.162±98.9,1548.668±101.0。瘤体平均瘤体重量(g)为:0.82±0.10,1.68±0.21,1.90±0.25。平均生长率为:18.711%,38.286%,39.709%。两组实验组分别和空载体阴性对照组、对照组比较有统计学意义(P<0.05),空载体阴性对照组和对照组比较无统计学意义(P>0.05)。结论:针对Bcl-2基因siRNA真核表达载体通过局部多点注射于移植瘤周围可以有效降低Bcl-2基因的表达,该基因沉默后肿瘤细胞增殖受到抑制,体内瘤体生长减慢。
Objective：Investigating RNA interference mediated Bcl-2 gene silence can inhibit growth of the cholangiocarcinoma and gallbladder carcinoma xenograft in nude mice.Methods： The animal models of human cholangiocarcinoma and Gallbladder carcinoma in nude mice body were set up.The nude mice were randomly divided into three groups： pSilencerTM-EGFP sh515,pSilencerTM-EGFP shCon,control group.After the formation of tumor knot,siRNA expressing plasmids pSilencerTM-EGFP sh515 and pSilencerTM-EGFP shCon were injected into the knots of pSilencerTM-EGFP sh515 group and pSilencerTM-EGFP shCon group.The volumes of the tumors were measured and the tumor growth rates were calculated.All mice were sacrificed after 6 weeks.The distribution of Bcl-2 in the tumors was detected by immunohistochemistry.Results： The nude mouse model of human cholangiocarcinoma and gallbladder cancer xenograft tumor was constructed successfully.The cholangiocarcinoma average tumor volumes（mm3）in pSilencerTM-EGFP sh515 group,pSilencerTM-EGFP shCon group,control group was 801.776±59.6,1383.980±78.2,1490.235±89.0,and the average tumor weight（g） was 0.90±0.11,1.49±0.31,1.63±0.22,and the average growth rates was 20.558%,35.587%,38.211%.The gallbladder cancer average tumor volumes（mm3）in pSilencerTM-EGFP sh515 group,pSilencerTM-EGFP shCon group,control group were 729.736±66.6,1493.162±98.9,1548.668±101.0,and the average tumor weight（g） was 0.82±0.10,1.68±0.21,1.90±0.25,and the average growth rates were 18.711%,38.286%,39.709%.The average tumor volume,weight and growth rates of pSilencerTM-EGFP sh515 group was significantly smaller than that of pSilencerTM-EGFP shCon group and control group（P0.05）,and there was no significant difference between pSilencerTM-EGFP shCon group and control group（P0.05）.Conclusion： By local injection,siRNA vector can effectively degrade the expressive level of Bcl-2 gene.After silence of Bcl-2 gene,the xenograft growth rate decreased.
出处
《现代肿瘤医学》
CAS
2012年第1期31-34,共4页
Journal of Modern Oncology
基金
陕西省自然科学基金资助项目(No.2003C2 47)
