PMID- 34939761
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20220429
IS  - 2228-5806 (Print)
IS  - 2228-5814 (Electronic)
IS  - 2228-5806 (Linking)
VI  - 23
IP  - 6
DP  - 2021 Nov
TI  - Restoration of MiR-34a Expression by 5-Azacytidine Augments Alimta -Induced Cell 
      Death in Non-Small Lung Cancer Cells by Downregulation of HMG B1, A2 and Bcl-2 
      Pathway.
PG  - 674-683
LID - 10.22074/cellj.2021.7332 [doi]
AB  - OBJECTIVE: Alimta (Pemetrexed) as an antifolate drug has been approved for the 
      treatment of lung cancer. The aim of the present study was to investigate the 
      combination effect of 5-Azacytidine (5-aza) and Alimta on the miR-34a and its 
      target genes expression and induction of apoptotic cell death in non-small lung 
      cancer A549 cells. MATERIALS AND METHODS: In this experimental study, lung cancer 
      A549 cells were treated with various concentrations of Alimta alone and combined 
      with 5-Aza. Then, viability was assessed by trypan blue and MTT assays. mRNA 
      expressions were performed by real time-polymerase chain reaction (PCR) and 
      western blot. Flow cytometry used to detect apoptotic/ necrotic cells and cell 
      cycle arrest. RESULTS: Alimta alone reduced viability of the cells in a dose 
      dependent manner with the half-maximal inhibitory concentration (IC(50)) value of 
      12 muM. Pretreatment of the cells with 5-aza (5 muM) induced a synergistic 
      cytotoxic effect with IC(50) of 3 muM. Sequential exposure of the cells to 5-aza 
      and Alimta enhanced miR-34a expression and significantly downregulated HMGB1, 
      HMGA2 and BCL-2 expressions. Also, it was associated with reduction of nuclear 
      HMGB1 and HMGA2 content. Caspase-3 activation, HMGB1 release into extracellular 
      space and staining of the cells with annexine V/PI suggested that 5-aza reduced 
      late apoptotic/necrotic cell death induced by Alimta. In addition, combination of 
      5-aza and Alimta arrested the cells at S and sub-G1 phases and inhibited colony 
      formation. CONCLUSION: 5-aza synergistically enhances Alimta induced apoptotic 
      cell death through HMG proteins regulation, MIR34A gene expression and intrinsic 
      apoptosis mechanism, providing a promising combination therapy in clinical lung 
      cancer therapy.
CI  - Copyright(c) by Royan Institute. All rights reserved.
FAU - Amiri, Somayeh
AU  - Amiri S
AD  - Department of Biochemistry, Institute of Biochemistry and Biophysics, University 
      of Tehran, Tehran, Iran.
FAU - Rabbani-Chadegani, Azra
AU  - Rabbani-Chadegani A
AD  - Department of Biochemistry, Institute of Biochemistry and Biophysics, University 
      of Tehran, Tehran, Iran. Email: arabbani@ut.ac.ir.
FAU - Davoodi, Jamshid
AU  - Davoodi J
AD  - Department of Biochemistry, Institute of Biochemistry and Biophysics, University 
      of Tehran, Tehran, Iran.
FAU - Gol Fakhrabadi, Hoda
AU  - Gol Fakhrabadi H
AD  - Department of Biochemistry, Institute of Biochemistry and Biophysics, University 
      of Tehran, Tehran, Iran.
LA  - eng
PT  - Journal Article
DEP - 20211123
PL  - Iran
TA  - Cell J
JT  - Cell journal
JID - 101566618
PMC - PMC8665979
OTO - NOTNLM
OT  - 5-Azacytidine
OT  - Alimta
OT  - Apoptosis
OT  - Lung Cancer Cell
OT  - miR-34a
COIS- There is no conflict of interest in this study.
EDAT- 2021/12/24 06:00
MHDA- 2021/12/24 06:01
PMCR- 2021/11/01
CRDT- 2021/12/23 08:55
PHST- 2019/12/21 00:00 [received]
PHST- 2020/03/16 00:00 [accepted]
PHST- 2021/12/23 08:55 [entrez]
PHST- 2021/12/24 06:00 [pubmed]
PHST- 2021/12/24 06:01 [medline]
PHST- 2021/11/01 00:00 [pmc-release]
AID - 10.22074/cellj.2021.7332 [doi]
PST - ppublish
SO  - Cell J. 2021 Nov;23(6):674-683. doi: 10.22074/cellj.2021.7332. Epub 2021 Nov 23.