CXCR4-shRNA纳米复合微粒对肾癌细胞增殖的抑制作用

发布时间：2018-06-26 14:29

本文选题：聚酰胺胺型树枝状分子 + CXCR　；参考：《中国肿瘤生物治疗杂志》2015年01期

【摘要】：目的:制备聚酰胺胺型树枝状分子(polyamidoamine dendrimer,PAMAM-D)载送CXCR4-shRNA的纳米复合微粒(PAMAM-shRNA),研究其在体外抑制人肾癌A498细胞增殖的效应。方法:将第7代的PAMAM-D室温下与CXCR4-shRNA混合(质量比为1∶0.73),制备成纳米树形分子与CXCR4-shRNA的纳米复合物PAMAM-shRNA,采用透射电镜观察PAMAMshRNA的形态结构,激光粒径仪测定其粒径。分别以PAMAM-shRNA、CXCR4-shRNA和PAMAM-D转染A498细胞,MTT法检测PAMAM-shRNA对肾癌细胞A498增殖的抑制作用,流式细胞术检测PAMAM-shRNA诱导A498细胞的凋亡情况,Real-time PCR分析转染后肾癌A498细胞CXCR4 mRNA的表达水平。结果:成功制备的新型纳米复合微粒PAMAM-shRNA分散性好,不粘连,其平均粒径为(176.5±25.48)nm。PAMAM-shRNA可以有效地抑制人肾癌细胞A498的增殖,且随着PAMAM-shRNA浓度和药物作用时间的增加,细胞增殖抑制的效果越明显,其最高增殖抑制率达(66.5±2.7)%;其还可加强诱导肾癌A498细胞凋亡。Real-time PCR检测结果表明,与CXCR4-shRNA组相比,PAMAM-shRNA组的CXCR4 mRNA的表达水平明显下降[(0.29±0.035)vs(0.70±0.084),P0.05]。结论:PAMAM-D能高效递送CXCR4-shRNA进入A498细胞,PAMAM-shRNA以剂量和时间依赖方式显著抑制肾癌细胞增殖和诱导肾癌细胞凋亡,其在肿瘤基因治疗中具有潜在的应用价值。
[Abstract]:Aim: to prepare polyamidoamine dendrimer-PAMAM-D nanoparticles (PAMAM-shRNA) carrying CXCR4-shRNA, and to study the inhibitory effect of PAMAM-shRNA on the proliferation of human renal cell carcinoma A498 cells in vitro. Methods: the 7th generation PAMAM-D was mixed with CXCR4-shRNA at room temperature (mass ratio 1: 0.73), and the nanocomposite PAMAM-shRNA was prepared. The morphology of PAMAM-shRNA was observed by transmission electron microscope. The particle size of PAMAMshRNA was measured by laser particle size analyzer. The inhibitory effect of PAMAM-shRNA on the proliferation of renal cancer cell A498 was detected by MTT assay with PAMAM-shRNA-CXCR4-shRNA and PAMAM-D transfected A498 cell line. Flow cytometry was used to detect the apoptosis of A498 cells induced by PAMAM-shRNA. Real-time PCR was used to analyze the expression level of CXCR4 mRNA in A498 cells transfected with PAMAM-shRNA. Results: PAMAM-shRNA, a new type of nano-composite particle, had good dispersity and no adhesion. The average diameter of PAMAM-shRNA was (176.5 卤25.48) nm.PAMAM-shRNA could effectively inhibit the proliferation of human renal cancer cell A498, and with the increase of PAMAM-shRNA concentration and drug action time, PAMAM-shRNA could effectively inhibit the proliferation of human renal cancer cell line A498. The more obvious the effect of cell proliferation inhibition was, the highest inhibitory rate of cell proliferation was (66.5 卤2.7), and the results of real-time PCR showed that the expression level of CXCR4 mRNA in PAMAM-shRNA group was significantly lower than that in CXCR4-shRNA group [(0.29 卤0.035) vs (0.70 卤0.084) P 0.05], and the expression of CXCR4 mRNA in CXCR4-shRNA group was significantly lower than that in CXCR4-shRNA group [(0.29 卤0.035) vs (, 0.70 卤0.084) (P0.05)]. Conclusion: the fraction PAMAM-D can efficiently deliver CXCR4-shRNA into A498 cells and inhibit the proliferation and apoptosis of RCC cells in a dose-and time-dependent manner. It has potential application value in tumor gene therapy.
【作者单位】：武警上海总队医院肾内科;第二军医大学长海医院特需诊疗科;
【基金】：国家自然科学基金资助项目(No.81202019) 上海市卫生局青年科研资助项目(No.2011Y197)~~
【分类号】：R737.11

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