Keshavarz Reihaneh, Bakhshinejad Babak, Babashah Sadegh, Baghi Narges, Sadeghizadeh Majid
Department of Molecular Genetics, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran.
Iran J Basic Med Sci. 2016 Dec;19(12):1353-1362. doi: 10.22038/ijbms.2016.7923.
Glioblastoma is the most lethal tumor of the central nervous system. Here, we aimed to evaluate the effects of exogenous delivery of p53 and a nanoformulation of curcumin called dendrosomal curcumin (DNC), alone and in combination, on glioblastoma tumor cells.
MTT assay was exploited to measure the viability of U87-MG cells against DNC treatment. Cells were separately subjected to DNC treatment and transfected with p53-containing vector and then were co-exposed to DNC and p53 overexpression[A GA1][B2]. Annexin-V-FLUOS staining followed by flow cytometry and real-time PCR were applied to examine apoptosis and analyze the expression levels of the genes involved in cell cycle and oncogenesis, respectively.
The results of cell viability assay through MTT indicated that DNC inhibits the proliferation of U87-MG cells in a time- and dose-dependent manner. Apoptosis evaluation revealed that p53 overexpression accompanied by DNC treatment can act in a synergistic manner to significantly enhance the number of apoptotic cells (90%) compared with their application alone (15% and 38% for p53 overexpression and DNC, respectively). Also, real-time PCR data showed that the concomitant exposure of cells to both DNC and p53 overexpression leads to an enhanced expression of GADD45 and a reduced expression of NF-κB and c-Myc.
The findings of the current study suggest that our combination strategy, which merges two detached gene (p53) and drug (curcumin) delivery systems into an integrated platform, may represent huge potential as a novel and efficient modality for glioblastoma treatment.
胶质母细胞瘤是中枢神经系统中最致命的肿瘤。在此，我们旨在评估单独及联合外源性递送p53和一种名为树枝状姜黄素（DNC）的姜黄素纳米制剂对胶质母细胞瘤肿瘤细胞的影响。
采用MTT法检测U87 - MG细胞对DNC处理的活力。细胞分别接受DNC处理并转染含p53的载体，然后同时暴露于DNC和p53过表达[AG1][B2]。采用膜联蛋白V - FLUOS染色后进行流式细胞术检测凋亡情况，并通过实时PCR分别分析细胞周期和肿瘤发生相关基因的表达水平。
MTT法检测细胞活力的结果表明，DNC以时间和剂量依赖性方式抑制U87 - MG细胞的增殖。凋亡评估显示，与单独应用相比（p53过表达和DNC单独应用时凋亡细胞分别为15%和38%），p53过表达联合DNC处理可协同作用显著增加凋亡细胞数量（达90%）。此外，实时PCR数据表明，细胞同时暴露于DNC和p53过表达会导致GADD45表达增强，NF - κB和c - Myc表达降低。
本研究结果表明，我们将两个独立的基因（p53）和药物（姜黄素）递送系统整合到一个综合平台的联合策略，可能作为一种新型高效的胶质母细胞瘤治疗方式具有巨大潜力。
