靶向递送携带HIF-2α siRNA的阳离子脂质体抑制肾透明细胞癌生长

Targeted delivery carrying HIF-2αsiRNA cationic liposomes inhibit the growth of clear cell renal cell carcinoma

目的:探索靶向缺氧诱导因子-2α(hypoxia-inducible factor-2α,HIF-2α)的阳离子脂质体-siRNA复合物(liposome-siRNA,Lipo-siRNA)是否可以有效干扰HIF-2α的表达并抑制肾透明细胞癌的生长。方法:使用ZetaSizer Nano及电镜鉴定Lipo-siRNA,荧光显微镜观察脂质体介导的siRNA转染786-O细胞的情况,CCK-8检测Lipo-siRNA对786-O细胞的抑制能力,Western blot检测Lipo-siRNA对HIF-2α表达的影响,活体成像检测Lipo-FAM-siRNA的体内转染情况,裸鼠皮下移植瘤模型检测Lipo-siRNA对肿瘤生长的抑制情况。结果:Zata电位、粒径大小及电镜观测结果均表明Lipo-siRNA构建成功;荧光显微镜观察表明脂质体可以有效介导FAM-siRNA对786-O细胞的转染(P<0.05);CCK-8实验表明Lipo-siRNA可以显著沉默HIF-2α表达(P<0.01)并抑制786-O细胞的增殖(P<0.05);活体成像结果显示Lipo-siRNA可以显著富集于肿瘤部位(P<0.001);裸鼠皮下移植瘤模型显示Lipo-siRNA可以有效降低肿瘤组织中HIF-2α的表达(P<0.05),并抑制肿瘤生长(P<0.05)。结论:携带靶向HIF-2αsiRNA的阳离子脂质体可以在裸鼠体内外水平显著干扰HIF-2α的表达,抑制肾透明细胞癌细胞的生长,具有治疗肾透明细胞癌的潜力。

Objective:To explore whether the cationic liposome-siRNA complex(Lipo-siRNA)targeting hypoxia-inducible factor-2α(HIF-2α)could effectively interfere with the expression of HIF-2αand inhibit the growth of renal clear cell carcinoma.Methods:The liposomal-siRNA complex was identified by ZetaSizer Nano and electron microscopy.Liposome mediated siRNA transfection of 786-O was observed by fluorescence microscopy.CCK-8 was used to detect the inhibitory effect of Lipo-siRNA on 786-O cells.The effect of Lipo-siRNA on HIF-2αexpression was detected by Western blot.In vivo transfection of Lipo-FAM-siRNA was detected by in vivo imaging.The inhibition of Lipo-siRNA on tumor growth was detected in subcutaneous tumor transplantation model in nude mice.Results:The results of Zata potential,particle size and electron microscope observation showed that the liposomal-siRNA complex was successfully constructed.Fluorescence microscope observation showed that liposomes could effectively mediate the transfection of FAM-siRNA into 786-O cells(P<0.05).CCK-8 assay showed that Lipo-siRNA could significantly silenced HIF-2αexpression(P<0.01)and inhibit 786-O cell proliferation(P<0.05).In vivo imaging results showed that Lipo-siRNA could significantly enrich in tumor sites(P<0.001).Subcutaneous xenograft tumormodel of nude mice showed that Lipo-siRNA could effectively reduce HIF-2αexpression(P<0.05)in tumor tissues and inhibit tumor growth(P<0.05).Conclusion:Cationic liposome Lipo-siRNA carrying HIF-2αsiRNA can significantly inhibit HIF-2αexpression in vitro and in vivo,inhibit the growth of renal clear cell carcinoma cells,and has the potential to treat renal clear cell carcinoma.