联合转染Survivin和Bcl-2反义寡核苷酸对裸鼠胆囊癌生长的抑制作用

THE DEPRESSANT EFFECT OF COMBINING ANTISENSE OLIGONUCLEOTIDE TARGETING Survivin AND Bcl-2 ON GALLBLADDER CARCINOMA IN NUDE MOUSE

目的探讨联合转染Survivin和Bcl-2反义寡核苷酸(AsODN)对裸鼠胆囊癌生长抑制作用。方法建立裸鼠胆囊癌皮下移植瘤模型,选择36只裸鼠随机分为3组:对照组、正义寡核苷酸(sODN)组和AsODN组,裸鼠瘤体内分别注射PBS、Survivin和Bcl-2sODN、Survivin和Bcl-2AsODN,12d后处死裸鼠,测量瘤体体积及质量,计算抑瘤率;苏木精-伊红染色观察瘤体组织学变化;免疫组织化学方法检测Survivin及Bcl-2蛋白的表达。结果AsODN组裸鼠瘤体平均体积显著低于对照组、sODN组(F=24.469,q=8.802、7.569,P<0.01),其体积抑瘤率为55.53%,sODN组为9.90%;AsODN组裸鼠瘤体平均质量显著低于对照组、sODN组(F=25.376,q=3.168、20.737,P<0.01),其质量抑瘤率为56.24%,sODN组为8.47%。AsODN组瘤体中可见大量的凋亡细胞。免疫组化证实AsODN组Survivin表达明显降低,同sODN组相比差异有显著意义(F=15.103,q=5.883、7.191,P<0.01);Bcl-2表达亦明显降低,同sODN组相比差异有显著性(F=6.632,q=4.793、4.030,P<0.05)。结论联合转染Survivin和Bcl-2AsODN可下调Survivin和Bcl-2表达,诱发细胞凋亡,从而抑制裸鼠胆囊癌种植瘤的生长。

Objective To evaluate the inhibitory action of combining antisense oligonucleotide targeting Survivin and Bcl- 2 on growth of gallbladder carcinoma in nude mouse. Methods A model of human gallbladder carcinoma transplanted subcutaneously in nude mice was established. A total of 36 mice were divided into three groups： control group, Survivin and Bcl-2 sODN group, as well as survivin and Bel-2 AsODN group. Twelve days later, the weight and volume of the tumor were measured and histology observed by HE staining, the expression of survivin and Bcl-2 protein detected by immunohistochemistry SABC method. Results The mean volume and weight of the tumors of AsODN group were significantly lower than that of the controls and sODN group （F=24. 469,25. 376 ;q=3. 168-20. 737 ;P〈0. 01）. The inhibition rate of volume and weight of AsODN group was 55.53% and 56.24%, respectively, while that of sODN group was 9.90% and 8. 47%. A lot of apoptotic cells were seen in tissue sections with HE staining, and the expression of survivin and Bel-2 protein was markedly lower than that of the control group and sODN group （F=15. 103,6. 632 ;q=4. 030-7. 191;P〈0. 05）. Conclusion Combining antisense oligonucleotide targeting survivin and Bcl-2 could inhibit the growth of subcutaneous tumors in nude mice, the reason might be that they could down-regulate the expression of survivin and Bcl-2 and induce apoptosis of gallbladder carcinoma cell.