存活素与血管内皮生长因子反义寡核苷酸对人甲状腺髓样癌裸鼠模型的作用

目的探讨存活素与血管内皮生长因子（VEGF）反义寡核苷酸（ASODN）对人甲状腺髓样癌（MTC）裸鼠模型的作用。方法构建的28只人甲状腺髓样癌裸鼠模型，随机分配到对照组和治疗组。对照组包括未处理组，正义寡核苷酸（SODN）组（VEGF SODN组和存活素SODN组）和SODN联合组；治疗组包括ASODN组（VEGF ASODN组和存活素ASODN组）和ASODN联合组。各组每日一次瘤内注射相应处理液，连续7d。治疗结束后l周末，杀死全部瘤鼠，切取瘤体，测算瘤重和抑瘤率以评价肿瘤生长情况，瘤组织石腊切片分别行HE、免疫组化和DNA末端原位标记法（TUNEL法）染色以评价组织病理构造、VEGF与存活素蛋白表达和细胞凋亡的变化。结果HE染色显示，治疗组血管生成明显降低。此外，相对于对照组，治疗组具有显著的VEGF和存活素低表达、高凋亡、低瘤重和高抑瘤率特点（均P〈0．01），并且ASODN联合组较ASODN组更为显著（均P〈0．05），而对照组间和SODN组间未见上述各特点的统计学差异。结论在裸鼠MTC模型，人甲状腺髓样癌存活素和VEGF的单或双靶向基因沉默治疗能够达到明显抑瘤效果。并且联合治疗更为突出。这为发展甲状腺髓样癌的基因治疗提供了实验依据。

Effects of survivin and VEGF antisense oligodeoxyribonucleotides on human medullary thyroid cancer inoculated into nude mice

ObjectiveTo evaluate effects of survivin and vascular endothilsal growth factor (VEGF) antisense oligodeoxyribonucleotide (ASODN) on human medullary thyroid cancer (MTC) inoculated into nude mice. MethodsTwenty-eight nude mouse models bearing human MTC were constructed and randomly assigned to control groups and therapeutic groups. Control groups included untreated group, sense oligodeoxyribonucleotide (SODN) groups (VEGF SODN group and survivin SODN group) and combined SOND group; therapeutic groups included ASODN groups (VEGF ASODN group and survivin ASODN group) and combined ASODN group. Each group was given an intratumoral injection with serum-free F12 or corresponding transfection medium every day for seven days. All nude mice were sacrificed one week after termination of treatment; tumors were fully resected from each mouse and measured for weight and inhibitory rate (IR) of tumor growth; paraffin sections of each tumor were respectively stained with HE, immunohistochemical or Tdt-mediated dUTP nick end labeling (TUNEL) method to evaluate the variation of histopathological construction, VEGF and survivin protein expressions, and apoptosis. ResultsTherapeutic groups presented a trait of obviously depressed angiogenesis by HE staining. Compared with control groups, each therapeutic group possessed significant traits of lowered expressions of VEGF and survivin proteins, raised apoptosis, lowered tumor weight, and raised inhibitory rate (all P<0.01), furthermore, combined ASODN group showed even more significant changes than either of ASODN groups regrading those traits (all P<0.05), but no significant difference was found between individual control groups or ASODN groups regarding those traits (all P>0.05). ConclusionSurvivin and VEGF targeting single or double genetic silence therapy in MTC model of nude mice could achieve significant inhibitory effect on tumor growth, moreover, the combined one showed more marked effect,thus providing experimental proof for developing gene therapy of MTC.