端粒酶反义寡核苷酸基因对裸鼠体内膀胱癌生长的抑制作用

目的:探讨端粒酶反义寡核苷酸(ASON)基因对裸鼠体内膀胱癌生长的抑制作用,阐明反义端粒酶基因对膀胱癌的治疗作用。方法:膀胱癌EJ细胞株分别采用换液培养和用硫代磷酸修饰的反义寡核苷酸(PS-ASON)处理,将换液培养及PS-ASON处理的EJ细胞分别接种BALB/c裸鼠(对照组24只和PS-ASON处理组12只),观察致瘤率;肿瘤形成至20mm后将对照组裸鼠随机分为3组(n=8),隔日注射生理盐水(空白对照组)、PS-ASON(PS-ASON注射组)及化疗药物表阿霉素(表阿霉素组)。观察各组裸鼠致瘤时间、裸鼠体质量及肿瘤体积的变化,计算各组裸鼠生存率;光镜下观察肿瘤组织形态学变化。结果:PS-ASON处理组裸鼠致瘤率低于对照组(P<0.01),致瘤时间长于对照组(P<0.01),裸鼠平均体质量高于对照组(P<0.01)。按照不同分组,裸鼠注射不同药物治疗12d后,PS-ASON注射组和表阿霉素组裸鼠肿瘤体积均小于空白对照组(P<0.01);PS-ASON注射组与表阿霉素组裸鼠肿瘤体积比较差异无统计学意义(P>0.05)。空白对照组裸鼠体质量持续下降;接种16d后,PS-ASON注射组裸鼠体质量明显高于空白对照组(P<0.01);接种肿瘤8d后,表阿霉素组裸鼠体质量低于PS-ASON注射组(P<0.01);空白对照组裸鼠体质量与表阿霉素组比较差异无统计学意义(P>0.05)。光镜下观察,PS-ASON注射组和表阿霉素组肿瘤细胞出现大量凋亡和坏死。PS-ASON注射组裸鼠生存率明显高于空白对照组(P<0.001)和表阿霉素组(P<0.01)。结论:反义端粒酶基因对体内膀胱癌形成及生长有明显的抑制作用,抑制程度与化疗药表阿霉素相当,可以用于膀胱癌治疗的进一步研究。

Inhibitory effect of antisense oligonucleotides of telomerase on growth of bladder cancer in vivo of nude mice

Objective To investigate the inhibitory effect of antisense oligonucleotides(ASON)of telomerase on the growth of bladder cancer in vivo of nude mice and to confirm its therapeutic effect on bladder cancer. Methods Bladder carcinoma EJ cells cultured with medium and thiophosphoric acid-modfied ASON PS-ASON were inoculated into BALB/c mice as control group(n=24)and PS-ASON treatment group(n=12).The rates of tumorgenesis were observed in two groups.The mice in control group were divided into three groups(n=8) depending on the different medicines fed to the mice as soon as 20mm tumor formed.The mice in blank control group,PS-ASON injection group and chemotherapy medicine group were injected with saine,PS-ASON,and pharmorubicin,respectively.The tumor growth,weight of nude mice,tumor volumes and the survival rate were measured and observed. Then the morphological changes of tumor tissue were observed under lightmicroscope.Results The rate of tumorgenesis of the mice in PS-ASON treatment group was lower than that in control group(P<0.01),the time of tumorgenesis in PS-ASON treatment group was more than that in control group(P<0.01)and the weight of the mice in treatment group was heavier than that in control group(P<0.01). After the mice were treated for 12d,the tumor volumes of the mice in PS-ASON injection group and pharmorubicin group were smaller than that in blank control group(P<0.01),but there was no significant difference in tumor volume between PS-ASON injection group and pharmorubicin group(P>0.05).The weight of the mice in blank control group lost obviously;the weight of the mice in PS-ASON injection group was bigger than that in blank control group(P<0.01)16dafter inoculation.The weight of the mice in pharmorubicin group were lower than that in PS-ASON injection group(P<0.01)8dafter inoculation;but there was no significant difference in the weight of the mice between blank control group and pharmorubicin group(P>0.05).Necrosis and apoptosis of incubating tumor cells were observed in PS-ASON injection group and pharmorubicin group under light microscope. The survival rate of the nude mice in PS-ASON group was higher than those in blank group(P<0.001)and pharmorubicin group(P <0.01).Conclusion ASON of telomerase inhibit the growth of bladder cancer significantly in vivo.Its therapeutic efficacy is similar to the chemotherapy medicine,and it can be used for the further research on therapy for bladder cancer.