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目的探讨c-FLIP反义寡核苷酸(c-FLIP ASODN)纳米粒(NP)对裸鼠体内人眼眶横纹肌肉瘤移植瘤生长的影响,评估纳米粒作为基因载体的可行性。方法皮下种植法建立裸鼠人眼眶横纹肌肉瘤动物模型,瘤体内分别注射c-FLIP反义寡核苷酸纳米粒(ASODN NP组)、未包裹的c-FLIP反义寡核苷酸(ASODN组)及生理盐水(NS组),观察肿瘤体积、组织形态学改变;应用蛋白质印迹分析法及免疫组化染色检测各组肿瘤组织c-FLIP的表达情况;应用TUNEL细胞凋亡检测试剂盒检测肿瘤组织细胞凋亡情况。结果与其他两组相比,ASODN NP组肿瘤的生长受到抑制;蛋白质印迹分析显示ASODN NP组和ASODN组的c-FLIP表达均较NS组减少(P<0.05);免疫组化显示c-FLIP表达于胞质,ASODN NP组c-FLIP阳性细胞较其余两组减少(P<0.05);ASODN NP组及ASODN组肿瘤组织凋亡细胞增多,ASODN NP组更加明显,NS组仅见个别细胞凋亡。结论 c-FLIP ASODN NP可有效抑制人眼眶横纹肌肉瘤裸鼠移植瘤的生长。NP可作为一种安全有效的ASODN导入载体。
Objective To investigate the effect of c-FLIP ASODN nanoparticles on the growth of human orbital rhabdomyosarcoma xenografts in nude mice and to evaluate the feasibility of nanoparticles as a gene carrier. Methods Animal models of orbital rhabdomyosarcoma in nude mice were established by subcutaneous implantation. C-FLIP antisense oligonucleotide nanoparticles (ASODN NP group), c-FLIP antisense oligonucleotide (ASODN group) ) And saline (NS group). The tumor volume and histomorphology were observed. The expression of c-FLIP in tumor tissue was detected by Western blot and immunohistochemistry. TUNEL assay was used to detect the tumor Tissue apoptosis. Results Compared with the other two groups, the growth of tumor in ASODN NP group was inhibited. Western blot analysis showed that the expression of c-FLIP in ASODN NP group and ASODN group was lower than that in NS group (P <0.05). Immunohistochemistry showed that c-FLIP The expression of c-FLIP positive cells in ASODN NP group was lower than that in the other two groups (P <0.05). The number of apoptotic cells in ASODN NP group and ASODN group was more than that in ASODN NP group, but only in NS group . Conclusion c-FLIP ASODN NP can effectively inhibit the growth of human orbital rhabdomyosarcoma xenografts in nude mice. NP can be used as a safe and effective ASODN import vector.