Abstract
BACKGROUND. Bcl-2 is anti-apoptotic and overexpression is associated with prostate tumor aggressiveness. We hypothesized that Bcl-2 has a role in prostate cancer radiation (RT) response. The relationship of Bcl-2 expression in four prostate cancer cell lines, and the effect of modulating expression with a Bcl-2 antisense oligonucleotide (G3139, Genasense®, oblimersen sodium, Genta Incorporated), to RT was examined. METHODS. The four cell lines studied were LNCaP (wild type-p53), PCS (p53 null), Bcl-2 stably transfected LNCaP (LNCaP-BST), and Bcl-2 stably transfected PCS (PC3-BST) cells. Cells were treated with antisense (AS) Bcl-2 alone or with RT (2-6 Gy). Following RT, cells were processed at 3-6 hr for Western blots, 18 hr for Annexin V staining and flow cytometric analysis, 24 hr for caspases 3 + 7 quantification by fluorometric assay, and immediately for clonogenic survival. RESULTS. AS caused a significant reduction in Bcl-2 expression in all cell lines. P53 expression was elevated following RT treatment in LNCaP and LNCaP-BST cells. P21 was increased by RT treatment in all cell lines. AS caused a significant increase in caspase 3 + 7 activity over the mismatch (MM) controls in all cell lines. When AS was combined with RT, caspase 3 + 7 activity was further increased significantly over all other groups in all cell lines. Moreover, AS + RT resulted in significantly reduced clonogenic survival over MM + RT, which was dampened in the Bcl-2 overexpressing lines. CONCLUSIONS. To our knowledge, these data demonstrate for the first time that a Bcl-2 specific AS oligonucleotide sensitizes prostate cancer cells to RT. p53 is not required for this effect.
| Original language | English |
|---|---|
| Pages (from-to) | 331-340 |
| Number of pages | 10 |
| Journal | Prostate |
| Volume | 65 |
| Issue number | 4 |
| DOIs | |
| State | Published - Dec 1 2005 |
| Externally published | Yes |
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Keywords
- Antisense Bcl-2
- Apoptosis
- Prostate cancer
- Radiation
ASJC Scopus subject areas
- Urology
Cite this
Antisense Bcl-2 sensitizes prostate cancer cells to radiation. / Mu, Zhaomei; Hachem, Paul; Pollack, Alan.
In: Prostate, Vol. 65, No. 4, 01.12.2005, p. 331-340.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Antisense Bcl-2 sensitizes prostate cancer cells to radiation
AU - Mu, Zhaomei
AU - Hachem, Paul
AU - Pollack, Alan
PY - 2005/12/1
Y1 - 2005/12/1
N2 - BACKGROUND. Bcl-2 is anti-apoptotic and overexpression is associated with prostate tumor aggressiveness. We hypothesized that Bcl-2 has a role in prostate cancer radiation (RT) response. The relationship of Bcl-2 expression in four prostate cancer cell lines, and the effect of modulating expression with a Bcl-2 antisense oligonucleotide (G3139, Genasense®, oblimersen sodium, Genta Incorporated), to RT was examined. METHODS. The four cell lines studied were LNCaP (wild type-p53), PCS (p53 null), Bcl-2 stably transfected LNCaP (LNCaP-BST), and Bcl-2 stably transfected PCS (PC3-BST) cells. Cells were treated with antisense (AS) Bcl-2 alone or with RT (2-6 Gy). Following RT, cells were processed at 3-6 hr for Western blots, 18 hr for Annexin V staining and flow cytometric analysis, 24 hr for caspases 3 + 7 quantification by fluorometric assay, and immediately for clonogenic survival. RESULTS. AS caused a significant reduction in Bcl-2 expression in all cell lines. P53 expression was elevated following RT treatment in LNCaP and LNCaP-BST cells. P21 was increased by RT treatment in all cell lines. AS caused a significant increase in caspase 3 + 7 activity over the mismatch (MM) controls in all cell lines. When AS was combined with RT, caspase 3 + 7 activity was further increased significantly over all other groups in all cell lines. Moreover, AS + RT resulted in significantly reduced clonogenic survival over MM + RT, which was dampened in the Bcl-2 overexpressing lines. CONCLUSIONS. To our knowledge, these data demonstrate for the first time that a Bcl-2 specific AS oligonucleotide sensitizes prostate cancer cells to RT. p53 is not required for this effect.
AB - BACKGROUND. Bcl-2 is anti-apoptotic and overexpression is associated with prostate tumor aggressiveness. We hypothesized that Bcl-2 has a role in prostate cancer radiation (RT) response. The relationship of Bcl-2 expression in four prostate cancer cell lines, and the effect of modulating expression with a Bcl-2 antisense oligonucleotide (G3139, Genasense®, oblimersen sodium, Genta Incorporated), to RT was examined. METHODS. The four cell lines studied were LNCaP (wild type-p53), PCS (p53 null), Bcl-2 stably transfected LNCaP (LNCaP-BST), and Bcl-2 stably transfected PCS (PC3-BST) cells. Cells were treated with antisense (AS) Bcl-2 alone or with RT (2-6 Gy). Following RT, cells were processed at 3-6 hr for Western blots, 18 hr for Annexin V staining and flow cytometric analysis, 24 hr for caspases 3 + 7 quantification by fluorometric assay, and immediately for clonogenic survival. RESULTS. AS caused a significant reduction in Bcl-2 expression in all cell lines. P53 expression was elevated following RT treatment in LNCaP and LNCaP-BST cells. P21 was increased by RT treatment in all cell lines. AS caused a significant increase in caspase 3 + 7 activity over the mismatch (MM) controls in all cell lines. When AS was combined with RT, caspase 3 + 7 activity was further increased significantly over all other groups in all cell lines. Moreover, AS + RT resulted in significantly reduced clonogenic survival over MM + RT, which was dampened in the Bcl-2 overexpressing lines. CONCLUSIONS. To our knowledge, these data demonstrate for the first time that a Bcl-2 specific AS oligonucleotide sensitizes prostate cancer cells to RT. p53 is not required for this effect.
KW - Antisense Bcl-2
KW - Apoptosis
KW - Prostate cancer
KW - Radiation
UR - http://www.scopus.com/inward/record.url?scp=28744451267&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=28744451267&partnerID=8YFLogxK
U2 - 10.1002/pros.20303
DO - 10.1002/pros.20303
M3 - Article
C2 - 16015611
AN - SCOPUS:28744451267
VL - 65
SP - 331
EP - 340
JO - Prostate
JF - Prostate
SN - 0270-4137
IS - 4
ER -