摘要
目的探讨RNA干扰Bcl-2基因表达对人胆囊癌细胞GBC-SD裸鼠体内成瘤能力和移植瘤生长的影响。方法本课题分为成瘤能力和治疗两个方面。成瘤能力:用针对Bcl-2基因小干扰RNA(siRNA)真核表达载体稳定转染的人胆囊癌细胞GBC-SD和人胆囊癌细胞GBC-SD分别制作裸鼠模型,观察移植瘤的生长情况,计算肿瘤成瘤率、生长体积、生长率。治疗实验:制作人胆囊癌细胞GBC-SD移植瘤模型,随机分为pSilencerTM-EGFPsh515组(实验组)、pSilencerTM-EGFPshCon组(空载体阴性对照组)和正常对照组。实验组用Bcl-2基因的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,正常对照组不作处理,观察移植瘤的生长情况,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①成功地建立裸鼠人胆囊癌GBC-SD细胞的动物模型;②成瘤能力:人胆囊癌细胞GBC-SD组和Bcl-2siRNA稳定转染细胞组成瘤率分别为100%、60%,瘤体平均体积分别为(1914.6±125.0)mm3、(629.7±78.9)mm3,平均生长率分别为45.58%、14.99%,瘤体重量分别为(2.24±0.33)g、(0.77±0.12)g,两组比较差异均有统计学意义(P<0.05);③治疗实验:实验组、空载体阴性对照组及正常对照组瘤体平均体积分别为(729.7±66.6)mm3、(1493.2±98.9)mm3、(1548.67±101.0)mm3,瘤体平均重量分别为(0.82±0.10)g、(1.68±0.21)g、(1.90±0.25)g,平均生长率分别为18.711%、38.286%、39.709%。实验组与空载体阴性对照组及正常对照组比较差异均有统计学意义(P<0.05),而空载体阴性对照组与正常对照组比较无明显差异。结论针对Bcl-2基因siRNA真核表达载体转染人胆囊癌GBC-SD细胞可以有效降低Bcl-2基因的表达,该基因沉默后肿瘤细胞增殖受到抑制,体内成瘤能力下降,瘤体生长减慢。
Objective To investigate whether RNA interference in mediated Bcl-2 gene silence can decrease the gallbladder carcinoma xenograft formation rate and inhibit growth of xenograft in nude mice. Methods In the xenograft formation ability group, the Bcl-2-siRNAs were transfected into the gallbladder carcinoma cell. Transfected gallbladder carcinoma and gallbladder carcinoma were injected into nude mice hypodermically to form a gallbladder carcinoma tumor. The formation rate and volume of the tumors were measured and the tumor growth rates were calculated. In the therapy group, the animal models of human gallbladder carcinoma in nude mouse body were constructed. The nude mice were randomly divided into pSilencer^TM-EGFP sh515 group, pSilencer^TM-EGFP shCon group, and normal control group. After the formation of tumor knot, siRNA expressing plasmids pSilencer^TM-EGFP sh515 and pSilencer^M-EGFP shCon were injected into the knots of pSilencer^TM-EGFP sh515 group and pSilencer^TM-EGFP shCon group. The tumor volume was measured and tumor growth rate was calculated. Pall 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 gallbladder cancer xenograft tumor was constructed successfully.Xenograft formation ability： The xenograft formation rate of gallbladder carcinoma and Bcl-2 siRNA transfected gallbladder carcinoma was 100% and 60%, respectively; the average tumor volume in gallbladder carcinoma group and transfected gallbladder carcinoma group was （1 914.6 ± 125.0）mm^3 and （629.7±78.9） mm^3; the growth rate was 45.58% and 14.99%, and the average tumor weight was （2.24±0.33）g and （0.77±0.12）g, respectively. The tumor volume, weight and growth rate in the transfected gallbladder carcinoma group decreased significantly when compared with those in control group（P〈0.05）. Therapy group： The average tumor volume in pSilencerTM-EGFP sh515 group, pSilencerTM-EGFP shCon group and control group was （729. 7 ± 66. 6）, （1 493. 2±98.9） and （1 548.67±101.0）mm^3 ; the average tumor weight was （0.82±0.10）g, （1. 685±0.21）g and （1.90±0.25）g; the average growth rates was 18. 711%, 38. 286% and 39. 709%, respectively. The average tumor volume, weight and growth rate of pSilencerTM-EGFP sh515 group were significantly smaller than those in pSilencerTM-EGFP shCon group and normal control group （P〈0.05）, and there was no significant difference between pSilencerTM-EGFP shCon group and normal control group. Conclusion Gallbladder carcinoma GBC-SD cells infected with siRNA vector can effectively lower the level of Bcl-2 gene expression. After silence of Bcl-2 gene, the proliferation of GBC-SD cells is markedly inhibited, and the xenograft formation rate and growth rate are decreased significantly.
出处
《西安交通大学学报（医学版）》
CAS
CSCD
北大核心
2009年第1期93-96,共4页
Journal of Xi’an Jiaotong University（Medical Sciences）
基金
陕西省自然科学基金资助项目(No.2003C247)~~
