壳聚糖/shRNA PDGF-D纳米颗粒在乳腺癌中的体外基因沉默作用

Ceyda Ekentok, S. Turan, J. Akbuğa
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引用次数: 3

摘要

乳腺癌是世界上最常见的女性癌症,它是高度恶性和致命的。PDGF-D在许多细胞过程中起调节作用,如血管生成。PDGF-D在许多类型的癌症中过度表达,促进肿瘤生长和转移。利用shRNA和合适的载体系统沉默PDGF-D基因可能会抑制肿瘤的生长和转移。在我们的研究中,我们制备了负载五种不同shRNA质粒的壳聚糖纳米颗粒,靶向PDGF-D基因的不同外显子。然后,在体外对纳米颗粒进行了表征,并研究了这些纳米颗粒在乳腺癌细胞系(MCF- 7、MDA-MB-231和MDA-MB-435)中的转染效率。研究了单、多shRNA序列、壳聚糖分子量(150 kDa和400 kDa)、shRNA用量(100和500 μg)对纳米颗粒表征和转染效率的影响。纳米颗粒的尺寸在200 ~ 400 nm之间变化,封装效率约为95 ~ 100%。壳聚糖的分子量改变了shRNA的释放量。结果表明,含有靶向PDGF-D外显子6 (NP1) shRNA质粒的制剂在MDA-MB-231细胞株中具有最高的沉默效率。壳聚糖可作为靶向PDGF-D的shRNA基因传递系统。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
In vitro gene silencing effect of chitosan/shRNA PDGF-D nanoparticles in breast cancer
Breast cancer is the most common cancer worldwide in women and it is highly malignant and fatal. PDGF-D plays role in regulation of many cellular processes such as angiogenesis. PDGF-D is overexpressed in many types of cancers and promote tumor growth and metastasis. Silencing of PDGF-D gene by using shRNA with an appropriate carrier system may decrease tumor growth and metastasis. In our study, we prepared chitosan nanoparticles loaded with five different shRNA plasmids targeting different exons of PDGF-D gene. Then, nanoparticles were characterized in vitro and transfection efficiency of these nanoparticles were investigated in breast cancer cell lines (MCF- 7, MDA-MB-231 and MDA-MB-435). The effects of single and multiple shRNA sequences, molecular weight of chitosan (150 kDa and 400 kDa) and the amount of shRNA (100 and 500 μg) on the characterization and transfection efficiencies of nanoparticles have been studied. Size of nanoparticles changed between 200-400 nm and approximately 95-100% encapsulation efficiency were obtained. Release of shRNA changed with the molecular weight of chitosan. It was obtained that formulation containing shRNA plasmid targeting PDGF-D exon 6 (NP1) has the highest silencing efficiency in MDA-MB-231 cell line. It was also evaluated that chitosan can be a suitable gene delivery system for shRNA targeting PDGF-D.
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