Inhibition of potentially anti-apoptotic proteins by antisense protein kinase C-α (Isis 3521) and antisense bcl-2 (G3139) phosphorothioate oligodeoxynucleotides: Relationship to the decreased viability of T24 bladder and PC3 prostate cancer cells

L. Benimetskaya, P. Miller, S. Benimetsky, A. Maciaszek, P. Guga, S. L. Beaucage, A. Wilk, A. Grajkowski, A. L. Halperin, C. A. Stein

Research output: Contribution to journalArticle

Abstract

Isis 3521 and G3139 are 20- and 18-mer phosphorothioate oligonucleotides, respectively, targeted to the protein kinase C (PKC)-α and bcl-2 mRNAs. Treatment of T24 bladder and PC3 prostate carcinoma cells with full-length and 3′-truncation mutants of Isis 3521 causes down-regulation of PKC-α protein and mRNA. However, at the level of a 15-mer and shorter, down-regulation of mRNA expression is no longer observed. Further, no diminution in cellular viability, as measured by 3-(4,5-dimethylthiazol-2-yl)2,5-diphenyl tetrazolium bromide assay, in response to increasing concentrations of paclitaxel, can be observed for these shorter oligomers. These observations not only indicate that PKC-α protein expression can be down-regulated by both RNase H-dependent and -independent mechanisms but also that down-regulation of PKC-α is insufficient by itself to "chemosensitize" cells. G3139, which down-regulates bcl-2 protein and mRNA expression, also down-regulates PKC-α protein and mRNA expression but not that of PKC-βI, -ε, or -ζ. However, the down-regulation of PKC-α and bcl-2 are not linked. When the carrier Eufectin 5 is employed, only bcl-2 is down-regulated in both T24 and PC3 cells at 50 nM oligonucleotide concentration. At 100 nM, both bcl-2 and PKC-α expression are down-regulated, and only at this concentration can "chemosensitization" to paclitaxel and carboplatin be observed. In contrast, the down-regulation of bcl-2 seems to be linked with that of RelA (p65). However, this too is also not sufficient for chemosensitization, even though it leads to the loss of expression of genes under the putative control of nuclear factor-κB and to detachment of the cells from plastic surfaces. These results underscore the complexity of the intracellular requirements for the initiation of chemosensitization to anti-neoplastic agents.

Original languageEnglish (US)
Pages (from-to)1296-1307
Number of pages12
JournalMolecular Pharmacology
Volume60
Issue number6
StatePublished - 2001
Externally publishedYes

Fingerprint

Apoptosis Regulatory Proteins
Oligodeoxyribonucleotides
Protein Kinase C
Prostatic Neoplasms
Urinary Bladder
Down-Regulation
Messenger RNA
Paclitaxel
Proteins
Phosphorothioate Oligonucleotides
Ribonuclease H
oblimersen
Carboplatin
Oligonucleotides
Plastics
Prostate
Carcinoma
Gene Expression

ASJC Scopus subject areas

  • Pharmacology

Cite this

Inhibition of potentially anti-apoptotic proteins by antisense protein kinase C-α (Isis 3521) and antisense bcl-2 (G3139) phosphorothioate oligodeoxynucleotides : Relationship to the decreased viability of T24 bladder and PC3 prostate cancer cells. / Benimetskaya, L.; Miller, P.; Benimetsky, S.; Maciaszek, A.; Guga, P.; Beaucage, S. L.; Wilk, A.; Grajkowski, A.; Halperin, A. L.; Stein, C. A.

In: Molecular Pharmacology, Vol. 60, No. 6, 2001, p. 1296-1307.

Research output: Contribution to journalArticle

Benimetskaya, L. ; Miller, P. ; Benimetsky, S. ; Maciaszek, A. ; Guga, P. ; Beaucage, S. L. ; Wilk, A. ; Grajkowski, A. ; Halperin, A. L. ; Stein, C. A. / Inhibition of potentially anti-apoptotic proteins by antisense protein kinase C-α (Isis 3521) and antisense bcl-2 (G3139) phosphorothioate oligodeoxynucleotides : Relationship to the decreased viability of T24 bladder and PC3 prostate cancer cells. In: Molecular Pharmacology. 2001 ; Vol. 60, No. 6. pp. 1296-1307.
@article{2c0f02d247c34193b53ba45d9d4eb479,
title = "Inhibition of potentially anti-apoptotic proteins by antisense protein kinase C-α (Isis 3521) and antisense bcl-2 (G3139) phosphorothioate oligodeoxynucleotides: Relationship to the decreased viability of T24 bladder and PC3 prostate cancer cells",
abstract = "Isis 3521 and G3139 are 20- and 18-mer phosphorothioate oligonucleotides, respectively, targeted to the protein kinase C (PKC)-α and bcl-2 mRNAs. Treatment of T24 bladder and PC3 prostate carcinoma cells with full-length and 3′-truncation mutants of Isis 3521 causes down-regulation of PKC-α protein and mRNA. However, at the level of a 15-mer and shorter, down-regulation of mRNA expression is no longer observed. Further, no diminution in cellular viability, as measured by 3-(4,5-dimethylthiazol-2-yl)2,5-diphenyl tetrazolium bromide assay, in response to increasing concentrations of paclitaxel, can be observed for these shorter oligomers. These observations not only indicate that PKC-α protein expression can be down-regulated by both RNase H-dependent and -independent mechanisms but also that down-regulation of PKC-α is insufficient by itself to {"}chemosensitize{"} cells. G3139, which down-regulates bcl-2 protein and mRNA expression, also down-regulates PKC-α protein and mRNA expression but not that of PKC-βI, -ε, or -ζ. However, the down-regulation of PKC-α and bcl-2 are not linked. When the carrier Eufectin 5 is employed, only bcl-2 is down-regulated in both T24 and PC3 cells at 50 nM oligonucleotide concentration. At 100 nM, both bcl-2 and PKC-α expression are down-regulated, and only at this concentration can {"}chemosensitization{"} to paclitaxel and carboplatin be observed. In contrast, the down-regulation of bcl-2 seems to be linked with that of RelA (p65). However, this too is also not sufficient for chemosensitization, even though it leads to the loss of expression of genes under the putative control of nuclear factor-κB and to detachment of the cells from plastic surfaces. These results underscore the complexity of the intracellular requirements for the initiation of chemosensitization to anti-neoplastic agents.",
author = "L. Benimetskaya and P. Miller and S. Benimetsky and A. Maciaszek and P. Guga and Beaucage, {S. L.} and A. Wilk and A. Grajkowski and Halperin, {A. L.} and Stein, {C. A.}",
year = "2001",
language = "English (US)",
volume = "60",
pages = "1296--1307",
journal = "Molecular Pharmacology",
issn = "0026-895X",
publisher = "American Society for Pharmacology and Experimental Therapeutics",
number = "6",

}

TY - JOUR

T1 - Inhibition of potentially anti-apoptotic proteins by antisense protein kinase C-α (Isis 3521) and antisense bcl-2 (G3139) phosphorothioate oligodeoxynucleotides

T2 - Relationship to the decreased viability of T24 bladder and PC3 prostate cancer cells

AU - Benimetskaya, L.

AU - Miller, P.

AU - Benimetsky, S.

AU - Maciaszek, A.

AU - Guga, P.

AU - Beaucage, S. L.

AU - Wilk, A.

AU - Grajkowski, A.

AU - Halperin, A. L.

AU - Stein, C. A.

PY - 2001

Y1 - 2001

N2 - Isis 3521 and G3139 are 20- and 18-mer phosphorothioate oligonucleotides, respectively, targeted to the protein kinase C (PKC)-α and bcl-2 mRNAs. Treatment of T24 bladder and PC3 prostate carcinoma cells with full-length and 3′-truncation mutants of Isis 3521 causes down-regulation of PKC-α protein and mRNA. However, at the level of a 15-mer and shorter, down-regulation of mRNA expression is no longer observed. Further, no diminution in cellular viability, as measured by 3-(4,5-dimethylthiazol-2-yl)2,5-diphenyl tetrazolium bromide assay, in response to increasing concentrations of paclitaxel, can be observed for these shorter oligomers. These observations not only indicate that PKC-α protein expression can be down-regulated by both RNase H-dependent and -independent mechanisms but also that down-regulation of PKC-α is insufficient by itself to "chemosensitize" cells. G3139, which down-regulates bcl-2 protein and mRNA expression, also down-regulates PKC-α protein and mRNA expression but not that of PKC-βI, -ε, or -ζ. However, the down-regulation of PKC-α and bcl-2 are not linked. When the carrier Eufectin 5 is employed, only bcl-2 is down-regulated in both T24 and PC3 cells at 50 nM oligonucleotide concentration. At 100 nM, both bcl-2 and PKC-α expression are down-regulated, and only at this concentration can "chemosensitization" to paclitaxel and carboplatin be observed. In contrast, the down-regulation of bcl-2 seems to be linked with that of RelA (p65). However, this too is also not sufficient for chemosensitization, even though it leads to the loss of expression of genes under the putative control of nuclear factor-κB and to detachment of the cells from plastic surfaces. These results underscore the complexity of the intracellular requirements for the initiation of chemosensitization to anti-neoplastic agents.

AB - Isis 3521 and G3139 are 20- and 18-mer phosphorothioate oligonucleotides, respectively, targeted to the protein kinase C (PKC)-α and bcl-2 mRNAs. Treatment of T24 bladder and PC3 prostate carcinoma cells with full-length and 3′-truncation mutants of Isis 3521 causes down-regulation of PKC-α protein and mRNA. However, at the level of a 15-mer and shorter, down-regulation of mRNA expression is no longer observed. Further, no diminution in cellular viability, as measured by 3-(4,5-dimethylthiazol-2-yl)2,5-diphenyl tetrazolium bromide assay, in response to increasing concentrations of paclitaxel, can be observed for these shorter oligomers. These observations not only indicate that PKC-α protein expression can be down-regulated by both RNase H-dependent and -independent mechanisms but also that down-regulation of PKC-α is insufficient by itself to "chemosensitize" cells. G3139, which down-regulates bcl-2 protein and mRNA expression, also down-regulates PKC-α protein and mRNA expression but not that of PKC-βI, -ε, or -ζ. However, the down-regulation of PKC-α and bcl-2 are not linked. When the carrier Eufectin 5 is employed, only bcl-2 is down-regulated in both T24 and PC3 cells at 50 nM oligonucleotide concentration. At 100 nM, both bcl-2 and PKC-α expression are down-regulated, and only at this concentration can "chemosensitization" to paclitaxel and carboplatin be observed. In contrast, the down-regulation of bcl-2 seems to be linked with that of RelA (p65). However, this too is also not sufficient for chemosensitization, even though it leads to the loss of expression of genes under the putative control of nuclear factor-κB and to detachment of the cells from plastic surfaces. These results underscore the complexity of the intracellular requirements for the initiation of chemosensitization to anti-neoplastic agents.

UR - http://www.scopus.com/inward/record.url?scp=0035213894&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0035213894&partnerID=8YFLogxK

M3 - Article

C2 - 11723237

AN - SCOPUS:0035213894

VL - 60

SP - 1296

EP - 1307

JO - Molecular Pharmacology

JF - Molecular Pharmacology

SN - 0026-895X

IS - 6

ER -