RNA干扰Bcl-2基因表达对人胆管癌细胞及胆囊癌细胞株裸鼠移植瘤生长的抑制

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 Bel - 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: pSilencer? - EGFP sh515 ,pSilencer? - EGFP shCon,control group. After the formation of tumor knot, siRNA expressing plasmids pSilencerTM - EGFP sh515 and pSilencer - 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 Bel-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( mm )in pSilencer - 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 vol-umes( mm )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 pSilencer? - EGFP sh515 group was significantly smaller than that of pSilencer? - EGFP shCon group and control group ( P < 0. 05 ), and there was no significant difference between pSilencerTM - EGFP shCon group and control group ( P >0. 05 ). Conclusion: By local injection,siRNA vector can effectively degrade the expressive level of Bel - 2 gene. After silence of Bel - 2 gene, the xenograft growth rate decreased.