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目的:以水通道蛋白(AQP1)为靶点构建中性脂质体1,2-二油酰基磷脂酰胆碱(DOPC)介导的PGCsiRNA-AQP1重组质粒,转染至喉癌荷瘤小鼠体内,诱导RNA干扰(RNAi)沉默AQP1表达,探讨其对喉癌生长的抑制作用。方法:建立喉癌BALB/c小鼠移植模型,随机分为尾静脉及瘤周注射组、阴性及空白对照组(n=5),重组质粒转入小鼠瘤体,观察肿瘤细胞核固缩及凋亡,计算瘤体积及抑瘤率;免疫组织化学法检测瘤体内AQP1表达和微血管密度。结果:动物模型建立成功。1苏木精-伊红染色示尾静脉和瘤周注射组可见核固缩及凋亡小体,出现大量凋亡细胞;22个注射组的瘤体体积均小于对照组(P<0.01);尾静脉和瘤周注射组抑瘤率分别为52.4%和53.5%(P>0.05);3免疫组织化学法示2个注射组AQP1阳性细胞表达率和微血管密度均小于对照组(P<0.01)。结论:由DOPC介导能够将PGCsiRNA-AQP1重组质粒转入BALB/c小鼠瘤体内,并有效诱导RNAi,抑制小鼠体内喉癌组织中的AQP1及微血管生成,从而抑制喉癌的生长和发展。
OBJECTIVE: To construct a recombinant plasmid of PGCsiRNA-AQP1 mediated by neutral liposome 1,2-dioleoylphosphatidylcholine (DOPC) targeting aquaporin (AQP1) and transfect it into tumor-bearing mice of larynx In vivo, the expression of AQP1 was silenced by RNA interference (RNAi), and its inhibitory effect on the growth of laryngeal carcinoma was also investigated. Methods: BALB / c mice model of laryngeal carcinoma was established and randomly divided into tail vein and peritumoral injection group, negative control group and blank control group (n = 5). The recombinant plasmids were transplanted into the tumor of mice, The tumor volume and tumor inhibition rate were calculated. AQP1 expression and microvessel density were detected by immunohistochemistry. Results: The animal model was established successfully. 1 Hematoxylin-eosin staining showed nuclear condensation and apoptotic bodies in the tail vein and peritumoral injection group, and a large number of apoptotic cells appeared. The volume of tumor in the 22 injection groups was smaller than that in the control group (P <0.01). The inhibitory rates of AQP1 positive cells and microvessel density in the two injection groups were significantly lower than those in the control group (P <0.01) by immunohistochemical staining. The tumor inhibition rates of tail vein and peri-tumor injection group were 52.4% and 53.5% . CONCLUSION: DOPC-mediated transfection of PGCsiRNA-AQP1 into BALB / c mice can effectively induce RNAi and inhibit AQP1 and micro-angiogenesis in mouse laryngeal carcinoma and thus inhibit the growth and development of laryngeal carcinoma .