| 摘要: |
| 目的 将树枝状聚合物聚酰胺-胺(PAMAM)包载可以表达具有抑制人恶性黑素瘤细胞A375增殖、侵袭和转移的miR-34a的质粒sh-miR-34a,构建聚阳离子纳米复合物PAMAM/sh-miR-34a。考察形成的复合物的粒径、zeta电位、细胞摄取,以及对A375细胞的抑制作用。方法 利用粒度测定仪测定纳米复合物的粒径和电位,凝胶电泳实验测定PAMAM对sh-miR-34a的包裹能力; 利用罗丹明标记的sh-miR-34a考察A375细胞对纳米复合物的摄取;CCK8 法测定PAMAM/sh-miR-34a对A375细胞的抑制作用;Transwell 法测定PAMAM/sh-miR-34a抑制A375细胞迁移和侵袭作用;蛋白质印迹法测定PAMAM/sh-miR-34a对 A375细胞中pAkt、pRb、pERK1/2蛋白表达的阻滞作用。结果 PAMAM包裹sh-miR-34a可形成稳定的纳米复合物,在N/P=20时,细胞对PAMAM/sh-miR-34a的摄取达最高(P<0.05)。PAMAM可以有效携带sh-miR-34a进入A375细胞,体外抑制A375细胞的增殖、侵袭和迁移,并且可以阻滞A375细胞中pAkt、pRb和pERK1/2蛋白的表达。结论 PAMAM可以包裹sh-miR-34a并对人恶性黑素瘤A375细胞起到抑制作用。 |
| 关键词: 纳米复合物 聚酰胺-胺 miR-34a 黑素瘤 |
| DOI：10.16781/j.0258-879x.2016.02.0191 |
| 投稿时间：2015-05-30修订日期：2015-07-29 |
| 基金项目: |
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| Inhibitory effect of PAMAM/sh-miR-34a nanocomplexes on malignant melanoma cells |
| LI Jia-fei1,ZHANG Zi-jie1,CHEN Xiu-hui1,JI Tao2,WANG Yan-zong2* |
(1. Hebei North University, Zhangjiakou 075000, Hebei, China;
2. Health Cadre Training Center, PLA Beijing Military Area Command, Beijing 102300, China
*Corresponding author) |
| Abstract: |
| Objective To use polyamidoamine dendrimers (PAMAM) loaded with sh-miR-34a for constructing PAMAM/sh-miR-34a nanocomplexes and to observe its inhibitory effect on the proliferation, invasion and metastasis of malignant melanoma A375 cells. Methods Particle size analyzer was used to investigate the potential and size of the constructed PAMAM/sh-miR-34a nanocomplex, and sh-miR-34a enrichment capability was determined by gel electrophoresis assay. The intracellular uptake of PAMAM/sh-miR-34a by A375 cells was investigated using sh-miR-34a labeled by rhodamine. CCK8 method was used to determine the inhibitory effect of the nanocomplex against A375 cell proliferation. Transwell assay was used to determine the inhibition effect of nanocomplex against A375 cell migration and invasion. Western blotting analysis was used to examine the inhibitory effect of PAMAM/sh-miR-34a on the protein expression of pAkt, pRb, and pERK1/2 in A375 cells. Results PAMAM loaded with sh-miR-34a could form stable nanocomplexes. The intracellular uptake of PAMAM/sh-miR-34a by A375 cells was the highest when N/P=20 (P<0.05). PAMAM effectively mediated sh-miR-34a entry into A375 cells, inhibiting the cell proliferation, invasion and metastasis and blocking the protein expression of pAkt, pRb, and pERK1/2 in A375 cells. Conclusion PAMAM can enwrap sh-miR-34a and inhibit malignant melanoma A375 cells. |
| Key words: nanocomplex polyamidoamine dendrimers miR-34a melanoma |
