靶向端粒酶催化亚单位基因hEST2反义寡核苷酸对人肝癌细胞生长的抑制作用  

Inhibitory effect of antisense oligodeoxynucleofide targeted against telomerase catalytic subunit gene hEST2 on the growth of human hepatocellular carcinoma cells

作　　者：杨波[1] 脱朝伟[2] 张宁[2] 刘秋珍[2] 王明耀[3] 

机构地区：[1]解放军总医院南楼血液科,北京100853 [2]解放军第二0二医院电镜室,沈阳110003 [3]辽宁省肿瘤医院,沈阳110042

出　　处：《中华消化外科杂志》2007年第5期369-372,共4页Chinese Journal of Digestive Surgery

基　　金：国家“九五”重点科技攻关计划基金资助项目（96A230603）

摘　　要：目的观察靶向端粒酶催化亚单位基因hEST2反义寡核苷酸对人肝癌细胞生长的抑制作用。方法从Genebank中钓取人端粒酶催化亚单位基因hEST2的mRNA序列，通过计算机模建二级结构辅助设计并合成硫化修饰反义寡核苷酸（癌泰得）。采用人肝癌裸小鼠原位移植模型（HCM—Y89）。实验分为生理盐水组，5-Fu10mg／kg组，癌泰得50mg／kg组、75mg／kg组，癌泰得75mg／kg联合5-Fu10mg／kg组、50mg／kg联合5-Fu10mg／kg组。尾静脉给药，每天1次，连续20d。观察移植瘤质量和抑瘤率、移植瘤体积、AFP浓度和移植瘤组织学。结果（1）5-Fu10mg／kg组的抑瘤率为27．85％，其他各治疗组的抑瘤率均在42．14％～74．29％之间；（2）各治疗组瘤质量、瘤体积和AFP浓度均低于生理盐水组且差异有统计学意义；（3）瘤体积和AFP浓度之间呈直线关系，为显著正相关（r=0．9977）；（4）癌泰得和5-FU治疗人肝细胞癌后，肝癌细胞出现不同程度的凋亡、坏死，同时伴有纤维组织增生。结论癌泰得对肝癌细胞生长具有抑制作用，疗效优于5-FU；与5-FU合用具有协同抗肿瘤效应。Objective To investigate the inhibitory effect of antisense oligodeoxynueleotide targeted against telomerase catalytic subunit gene hEST2 on the growth of hepatoeellular carcinoma （HCC） cells. Methods mRNA sequence of hEST2 was obtained from Genebank database. Computer was applied in helping imitate the secondary structure of mRNA and design antisense oligodeoxynueleotide. Chemosynthesis method was used to synthesize the sulfurized antisense oligodeoxynueleotide targeted against hEST2 （Cantide）. Orthotopie implantation model of hepatoeellular carcinoma in nude mice （HCM-Y89） was established. All mice were divided into normal saline group （negative control） , 5-FU 10 mg/kg group （positive control） , Cantide 50 mg/kg group, Cantide 75 mg/kg group, Cantide 75 mg/kg plus 5-FU 10 mg/kg group, Cantide 50 mg/kg plus 5-FU 10 mg/kg group. Vena eaudalis injection was done on all mice once per day for 20 days continuously to observe weight, volume, histology and inhibition rate of implanted tumors, AFP concentration. Results （ 1 ） Inhibition rate of tumor was 27.85% in 5-FU 10 mg/kg group but 42.14%-74.29% in other groups. （2） In the treatment group, the tumor had lighter weight, smaller volume and lower AFP concentration than those in normal saline group. （3） AFP concentration had a positive correlation with tumor volume （ r =0.997 7）. （4） Apoptosis and necrosis of cancer cells and pro- liferation of fibrous tissue were observed after treating HCC by 5-FU and Cantide. Conclusions Cantide has an inhibitory effect on HCC and is much better than 5-FU in anti-tumor effect. Cantide combined with 5-FU has a synergistic anti-tumor effect.

关 键 词：端粒酶催化亚单位 反义寡核苷酸 原发性肝细胞癌 原位移植 裸小鼠 疾病模型 

分 类 号：R686[医药卫生—骨科学]