• Poly(butyl cyanoacrylate) nanoparticles treatment downregulates three proapoptotic BCL2 family genes in U87MG glioblastoma cell line
  • Mahdieh Sadat Taghavi,1,* Azim Akbarzadeh,2 Reza Mahdian,3
    1. Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran
    2. Pilot Biotechnology Department, Pasteur Institute of Iran, Tehran, Iran
    3. Biotechnology Research Center, Molecular Medicine Department, Pasteur Institute of Iran, Tehran, Iran


  • Introduction: Glioblastoma multiform is the most frequent malignant brain tumour. Brain tumours develop resistance to chemotherapy mainly due to the presence of the blood brain barrier (BBB) which blocks penetration of drugs into the brain. In the past few years, a number of different approaches have been developed for drugs to overcome this barrier. One of the most attractive methods is the use of polymeric nanoparticles. Poly(butyl cyanoacrylate) (PBCA), a biodegradable, biocompatible and minimally toxic polymer, has been proved to be promising candidate for CNS drug delivery. BAK, BAX and BAD are proapoptotic members of the BCL2 family that play key role in the regulation of apoptosis. In previous study we evaluated the toxicity of different concentrations of PBCA nanoparticles on U87MG cell lines. Our MTT assay and flow cytometry data indicate that within a concentration of 100μg/mL, empty PBCA nanoparticles imparts no cytotoxic effects on U87MG cell line. The objective of present study was to test the expression of BAK, BAX and BAD genes in U87MG cells treated with Poly(butyl cyanoacrylate) nanoparticles
  • Methods: Using emulsion polymerisation, the PBCA nanospheres were synthesised. Briefly, 1%(v/v) of butylcyanoacrylate was added dropwise to 1% (w/v) Dextran 70 solution (in 0.001 N HCl). The polymerisation was performed for 4 hours under constant stirring at 400 rpm and the mixture was neutralised and freeze-dried to obtain powder. Based on our pervious study, the non-toxic concentration of PBCA NPs on A172 cell line was determined as 100 μg/ml. Thus U87MG cell line were treated with 100 μg/ml of PBCA NPs for 48 hours. RNA was extracted and cDNA was synthesized. Gene expression study was performed on BAK, BAX and BAD as targets and TBP as internal control gene using very sensitive quantitative Real-time PCR.
  • Results: In real-time PCR assay, to confirm correct fragments amplification, melting curve analysis and gel electrophoresis were used. The target gene/TBP gene expression ratio was calculated for each gene using the formula 2−ΔΔCt . The ratios were calculated as 0.27 (p = 0.01), 0.10 (p = 0.01), 0.33 (p = 0.01) for BAX, BAD and BAK genes in U87MG cell line.
  • Conclusion: Gene expression analysis results showed that treatment with PBCA NPs at non-toxic dose caused significant changes in the expression of BAX, BAD and BAK genes in U87MG glioblastoma cell line. Global analysis of gene expression in different cell lines might provide more precise insight of molecular effects of Poly(butyl cyanoacrylate) nanoparticles on target cells.
  • Keywords: Poly(butyl cyanoacrylate) nanoparticles, apoptosis, BCL2 family