多功能纳米载体靶向传输Survivin siRNA抑制肝细胞癌增殖

Multifunctional Nanocarrier Targeting Survivin siRNA Inhibits Proliferation of Hepatocellular Carcinoma

【目的】研究RGD修饰、MRI可视的纳米载体(RGD-PEG-g-PEI-SPION)靶向传输Survivin siRNA至肝细胞癌抑制裸鼠肝癌生长的作用;探讨RGD-PEG-g-PEI-SPION对裸鼠肝癌的MRI显像功能。【方法】经皮下注射Bel-7402细胞构建裸鼠皮下荷瘤模型;经肿瘤生长抑制实验、肿瘤组织HE染色、免疫组织化学染色和肿瘤组织细胞凋亡试验评估RGD-PEG-g-PEI-SPION靶向传输Survivin siRNA对裸鼠肝癌的治疗功能;利用MRI成像分析RGD-PEG-g-PEI-SPION对肝细胞癌的靶向显像功能。【结果】裸鼠皮下荷瘤模型成功建立并经裸鼠尾静脉首次注射25 d后,RGD-PEG-g-PEI-SPION/siRNA、PEG-g-PEI-SPION/siRNA、RGD-PEG-g-PEI-SPION/siNC、PEG-g-PEI-SPION/siNC和PBS溶液裸鼠的皮下移植瘤体积分别为:59±8、156±7、202±7、212±9和220±8(mm3),肿瘤组织学检查显示RGD-PEG-g-PEI-SPION/siRNA实验组肿瘤坏死范围最广,Survivin蛋白表达量最低,凋亡的肿瘤细胞数量明显增多;在尾静脉注射RGD-PEG-g-PEI-SPION载体、PEG-g-PEI-SPION载体3 h后,较载体注射前皮下移植瘤的标准化MRI信号强度分别下降为原来的(51.6±4.3)%和(88.5±3.2)%。【结论】RGD-PEG-g-PEI-SPION靶向传输Survivin siRNA至肝细胞癌,促进肿瘤细胞凋亡,抑制肿瘤生长,取得了较好的抗肿瘤疗效;RGD-PEG-g-PEI-SPION对裸鼠肝癌组织具备良好的靶向MRI显像功能。

【Objective】To investigate the inhibitory effect of RGD-modified,MRI visualized nanocarrier(RGDPEG-g-PEI-SPION)targeting for Survivin siRNA(siRNA)delivery to hepatocellular carcinoma(HCC)in nude mice;and to investigate the MRI imaging function of RGD-PEG-g-PEI-SPION in nude mouse liver cancer.【Methods】Nude mice were subcutaneously injected with Bel-7402 cells to establish a subcutaneous tumor-bearing model.Tumor growth inhibition test,HE staining,immunohistochemical staining and tumor cell apoptosis test were used to evaluate the thera?peutic effect of RGD-PEG-g-PEI-SPION targeted Survivin siRNA on liver cancer in nude mouse;MRI imaging was used to analyze the targeting imaging function of RGD-PEG-g-PEI-SPION on hepatocellular carcinoma【Results】The subcu?taneous tumor-bearing model of nude mice was successfully established,and the tumor volume of subcutaneous graft of RGD-PEG-g-PEI-SPION/siRNA,PEG-g-PEI-SPION/siRNA,RGD-PEG-g-PEI-SPION/siNC,PEG-g-PEI-SPION/siNC and PBS solution in nude mice after first injection in the tail vein for 25 days was 59±8,156±7,202±7,212±9 and 220±8(mm3),respectively.Tumor histological examination showed that the RGD-PEG-g-PEI-SPION/siRNA group had the widest range of tumor necrosis,the lowest Survivin protein expression,and the number of apoptotic tumor cells signifi?cantly increased.Three hours after injection of RGD-PEG-g-PEI-SPION vector and PEG-g-PEI-SPION vector into the tail vein,the standardized MRI signal intensity of subcutaneous transplanted tumor decreased to(51.6±4.3)%and(88.5±3.2)%,respectively,compared with that before injection.【Conclusion】RGD-PEG-g-PEI-SPION delivers Survivin siRNA to hepatocellular carcinoma,promotes tumor cell apoptosis,inhibits tumor growth,and achieves good anti-tumor effect;RGD-PEG-g-PEI-SPION has a well targeted MRI imaging function in nude mouse liver cancer tissues.