Mellatyar Hassan, Akbarzadeh Abolfazl, Rahmati Mohammad, Ghalhar Masoud Gandomkar, Etemadi Ali, Nejati-Koshki Kazem, Zarghami Nosratallah, Barkhordari Amin
Department of Medical Biotechnology, Faculty of Advance Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran E-mail :
Asian Pac J Cancer Prev. 2014;15(20):8693-8. doi: 10.7314/apjcp.2014.15.20.8693.
Up-regulation of hsp90 gene expression occurs in numerous cancers such as lung cancer. D,L-lactic-co-glycolic acid-poly ethylene glycol-17-dimethylaminoethylamino-17-demethoxy geldanamycin (PLGA-PEG-17DMAG) complexes and free 17-DMAG may inhibit the expression. The purpose of this study was to examine whether nanocapsulating 17DMAG improves the anti cancer effect over free 17DMAG in the A549 lung cancer cell line.
Cells were grown in RPMI 1640 supplemented with 10% FBS. Capsulation of 17DMAG is conducted through double emulsion, then the amount of loaded drug was calculated. Other properties of this copolymer were characterized by Fourier transform infrared spectroscopy and H nuclear magnetic resonance spectroscopy. Assessment of drug cytotoxicity on the grown of lung cancer cell line was carried out through MTT assay. After treatment, RNA was extracted and cDNA was synthesized. In order to assess the amount of hsp90 gene expression, real-time PCR was performed.
In regard to the amount of the drug load, IC50 was significant decreased in nanocapsulated(NC) 17DMAG in comparison with free 17DMAG. This was confirmed through decrease of HSP90 gene expression by real-time PCR.
The results demonstrated that PLGA-PEG-17DMAG complexes can be more effective than free 17DMAG in down-regulating of hsp90 expression by enhancing uptake by cells. Therefore, PLGA-PEG could be a superior carrier for this kind of hydrophobic agent.
热休克蛋白90（hsp90）基因表达上调发生在众多癌症中，如肺癌。聚乳酸-乙醇酸共聚物-聚乙二醇-17-二甲氨基乙基氨基-17-去甲氧基格尔德霉素（PLGA-PEG-17DMAG）复合物和游离的17-DMAG可能会抑制其表达。本研究的目的是检验纳米包裹的17DMAG在A549肺癌细胞系中是否比游离的17DMAG具有更强的抗癌效果。
细胞在补充有10%胎牛血清的RPMI 1640培养基中培养。通过复乳法对17DMAG进行包封，然后计算载药量。通过傅里叶变换红外光谱和氢核磁共振光谱对该共聚物的其他性质进行表征。通过MTT法评估药物对肺癌细胞系生长的细胞毒性。处理后，提取RNA并合成cDNA。为了评估hsp90基因表达量，进行实时定量PCR。
就载药量而言，纳米包裹（NC）的17DMAG的半数抑制浓度（IC50）与游离的17DMAG相比显著降低。这通过实时定量PCR检测hsp90基因表达的降低得到证实。
结果表明，PLGA-PEG-17DMAG复合物通过增强细胞摄取，在下调hsp90表达方面可能比游离的17DMAG更有效。因此，PLGA-PEG可能是这类疏水性药物的优良载体。
