TY - JOUR

T1 - Downregulation of TdT Expression through Splicing Modulation by Antisense Peptide Nucleic Acid (PNA)

AU - Montazersaheb, Soheila

AU - Kazemi, Masoumeh

AU - Nabat, Elahe

AU - Nielsen, Peter E

AU - Hejazi, Mohammad S

N1 - Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.

PY - 2019

Y1 - 2019

N2 - BACKGROUND AND OBJECTIVE: Antisense oligonucleotides are able to modulate splicing patterns and offer therapeutic intervention for cancer and other diseases. Considering TdT potential as a target in cancer therapy, the present study aimed to investigate splicing alteration of TdT pre-mRNA in Molt-4 cells using peptide nucleic acid (PNA) octaarginine and cholic acid conjugates.METHOD: We examined 16 mer PNAs targeting 5' and 3' junctions of intron 7 and addressed their mRNA splicing modulation effects using RT-PCR analysis. We also tested corresponding 2-base mismatch PNAs to confirm the sequence specificity. In addition, protien level of TdT, apoptosis induction and cell viability rate were analysed.RESULTS: PCR analysis showed that full match PNAs could modulate the splicing process, thereby producing a longer mRNA still including intron 7. PCR results also implied exon 7 skipping. In addition, reduced level of TdT protein in Molt-4 cells was observed. Downregulation of TdT level in PNA treated cells was accompanied by an increased rate of apoptosis and decreased the level of cell survival.CONCLUSION: PNA-mediated splicing modulation can specifically downregulate TdT expression. TdT dowregulation results in apoptosis induction and reduced cell survival in Molt-4 cells. These observations could draw more attentions to develop PNA based strategies for TdT suppression and consequent apoptosis induction in acute lymphoblastic leukemia.

AB - BACKGROUND AND OBJECTIVE: Antisense oligonucleotides are able to modulate splicing patterns and offer therapeutic intervention for cancer and other diseases. Considering TdT potential as a target in cancer therapy, the present study aimed to investigate splicing alteration of TdT pre-mRNA in Molt-4 cells using peptide nucleic acid (PNA) octaarginine and cholic acid conjugates.METHOD: We examined 16 mer PNAs targeting 5' and 3' junctions of intron 7 and addressed their mRNA splicing modulation effects using RT-PCR analysis. We also tested corresponding 2-base mismatch PNAs to confirm the sequence specificity. In addition, protien level of TdT, apoptosis induction and cell viability rate were analysed.RESULTS: PCR analysis showed that full match PNAs could modulate the splicing process, thereby producing a longer mRNA still including intron 7. PCR results also implied exon 7 skipping. In addition, reduced level of TdT protein in Molt-4 cells was observed. Downregulation of TdT level in PNA treated cells was accompanied by an increased rate of apoptosis and decreased the level of cell survival.CONCLUSION: PNA-mediated splicing modulation can specifically downregulate TdT expression. TdT dowregulation results in apoptosis induction and reduced cell survival in Molt-4 cells. These observations could draw more attentions to develop PNA based strategies for TdT suppression and consequent apoptosis induction in acute lymphoblastic leukemia.

KW - Cell Line, Tumor

KW - DNA Nucleotidylexotransferase/genetics

KW - Down-Regulation

KW - Humans

KW - Oligonucleotides, Antisense/pharmacology

KW - Peptide Nucleic Acids/pharmacology

KW - RNA Splicing

U2 - 10.2174/1389201020666190206202650

DO - 10.2174/1389201020666190206202650

M3 - Journal article

C2 - 30727883

VL - 20

SP - 168

EP - 178

JO - Current Pharmaceutical Biotechnology

JF - Current Pharmaceutical Biotechnology

SN - 1389-2010

IS - 2

ER -