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An Effective Concentration of 5-Aza-CdR to Induce Cell Death and Apoptosis in Human Pancreatic Cancer Cell Line through Reactivating RASSF1A and Up-Regulation of Bax Genes | ||
| Iranian Journal of Medical Sciences | ||
| مقاله 10، دوره 43، شماره 5، آذر 2018، صفحه 533-540 اصل مقاله (966.65 K) | ||
| نوع مقاله: Original Article(s) | ||
| شناسه دیجیتال (DOI): 10.30476/ijms.2018.40574 | ||
| نویسندگان | ||
| Mehdi Nikbakht Dastjerdi1؛ Asaad Azarnezhad2؛ Batool Hashemibeni1؛ Mansour Salehi3؛ Mohammad Kazemi3؛ Zahra Babazadeh* 4 | ||
| 1Department of Anatomical Sciences, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran | ||
| 2Cellular and Molecular Research Center, Kurdistan University of Medical Sciences, Sanandaj, Iran; and Department of Medical Genetics, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran | ||
| 3Department of Molecular Biology, Isfahan University of Medical Science, Iran | ||
| 4Department of Anatomical Sciences, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran; and Department of Anatomical Sciences, Faculty of Medicine, Babol University of MedicalSciences, Babol, Iran | ||
| چکیده | ||
| Background: Promoter hyper-methylation of tumor suppressor genes is a common event that occurs in cancer. As methylation is a reversible modification, agents capable of reversing an abnormal methylation status should help to combat cancer. 5-Aza-CdR is a DNA methyl-transferase inhibitor. The present study aimed to evaluate the effect of 5-Aza-CdR on the proliferation of human pancreatic cancer cell line (PANC-1) and the expression of RASSF1A and Bax genes.Methods: PANC-1 cells were cultured and treated with 5 and 10 µM/L of 5-Aza-CdR for 24, 48, 72, and 96 hours and the percentages of cell viability and apoptosis were measured by MTT and flow cytometry. RASSF1A gene promoter methylation was assessed by methyl-specific primer-PCR (MSP-PCR) and the expression of RASSF1A and Bax genes was measured using quantitative real-time PCR (qPCR). All quantitative data are presented as mean±SD (standard deviation). The one-way analysis of variance (ANOVA) with the LSD post hoc test was performed for statistical analysis using the SPSS software package, version 16.0. Results: 3-[4,5-dimethythiaziazol-2yl]-2,5-diphenyl tetrazoliumbr omide (MTT) assay revealed that 5-Aza-CdR significantly inhibit the growth and proliferation of PANC-1. The flow cytometry results showed over 40% and 70% of early and late apoptotic cells after treatment with 5 and 10 µm/L of 5-Aza-CdR, respectively. MSP-PCR data indicated that the treatment of cells with 10 µm/L 5-Aza-CdR resulted in partial demethylation of RASSF1A gene promoter. qPCR results showed significant re-expression of RASSF1A and up-regulation of Bax genes after 96 hours treatment of cells with 10 µm/L 5-Aza-CdR versus control cells (P<0.01).Conclusion: The result demonstrated that 5 and 10 µM of 5-Aza-CdR induce cell death and apoptosis by epigenetic reactivation of RASSF1A and up-regulation of Bax genes. | ||
| کلیدواژه‌ها | ||
| Decitabine؛ Pancreatic Neoplasms؛ DNA Modification Methylases؛ Methylation | ||
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