Small molecule, oligonucleotide-based telomerase template inhibition in combination with cytolytic therapy in an in vitro androgen-independent prostate cancer model

Benjamin K. Canales, Yingming Li, Melissa G. Thompson, Joseph Gleason, Zhi Chen, Bahaa Malaeb, David R. Corey, Brittney Shea Herbert, Jerry W. Shay, Kenneth S. Koeneman

Research output: Contribution to journalArticle

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Abstract

Purpose: Determine the efficacy and timing of small molecule oligonucleotide-based inhibitors to the enzyme telomerase in an in vitro model of androgen-independent, osseous prostate cancer. Materials and Methods: Telomerase was inhibited in prostate cancer cell lines C4-2/C4-2B and in controls by using small molecule antisense oligonucleotide-based inhibitors alone or in various combinations of small-dose Taxotere® (sanofi-aventis, Bridgewater, NJ) and/or conditionally replication competent adenovirus (AD-BSP-E1a). After transfection and proliferation, telomerase telomeric repeat amplification protocol and telomere restriction fragment assays were performed, with specific times for evaluating telomere length. Specimens were stained for analysis with hematoxylin and eosin (H&E), terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling (TUNEL), and prostate-specific antigen (PSA). Results: C4-2/C4-2B cell lines had the shortest initial mean telomere length (approximately 2.5 kilobase [kb]) compared to PC-3 (approximately 5.5 kb). Dose-dependent inhibition of telomerase activity was seen using match oligonucleotide-based inhibitors to telomerase (50% inhibitory concentration 3-5 nm), whereas mismatch compound showed no telomerase inhibition. Significant growth delay and apoptosis in cell lines occurred after >50 days of treatment. Cells treated with combination "triple therapy" (i.e., telomerase inhibitors, adenovirus, and Taxotere®) had the highest amount of apoptosis. Compared to controls, all combination treatment groups had statistically significant reductions in prostate-specific antigen in the conditioned media. Conclusions: Combining cytotoxic regimens with small molecule inhibitors to telomerase with oligonucleotide-based agents could be beneficial in controlling osseous hormone refractory prostate cancer, as evidenced by these in vitro, preclinical investigations. Telomerase inhibition needs to move into in vivo models and human studies.

Original languageEnglish (US)
Pages (from-to)141-151
Number of pages11
JournalUrologic Oncology: Seminars and Original Investigations
Volume24
Issue number2 SPEC. ISS.
DOIs
StatePublished - Jan 1 2006
Externally publishedYes

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Telomerase
Oligonucleotides
Androgens
Prostatic Neoplasms
docetaxel
Telomere
Therapeutics
Prostate-Specific Antigen
Adenoviridae
Cell Line
Apoptosis
In Vitro Techniques
DNA Nucleotidylexotransferase
Antisense Oligonucleotides
Enzyme Inhibitors
Hematoxylin
Eosine Yellowish-(YS)
Conditioned Culture Medium
Inhibitory Concentration 50
Transfection

All Science Journal Classification (ASJC) codes

  • Oncology
  • Urology

Cite this

Small molecule, oligonucleotide-based telomerase template inhibition in combination with cytolytic therapy in an in vitro androgen-independent prostate cancer model. / Canales, Benjamin K.; Li, Yingming; Thompson, Melissa G.; Gleason, Joseph; Chen, Zhi; Malaeb, Bahaa; Corey, David R.; Herbert, Brittney Shea; Shay, Jerry W.; Koeneman, Kenneth S.

In: Urologic Oncology: Seminars and Original Investigations, Vol. 24, No. 2 SPEC. ISS., 01.01.2006, p. 141-151.

Research output: Contribution to journalArticle

Canales, Benjamin K. ; Li, Yingming ; Thompson, Melissa G. ; Gleason, Joseph ; Chen, Zhi ; Malaeb, Bahaa ; Corey, David R. ; Herbert, Brittney Shea ; Shay, Jerry W. ; Koeneman, Kenneth S. / Small molecule, oligonucleotide-based telomerase template inhibition in combination with cytolytic therapy in an in vitro androgen-independent prostate cancer model. In: Urologic Oncology: Seminars and Original Investigations. 2006 ; Vol. 24, No. 2 SPEC. ISS. pp. 141-151.
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AU - Canales, Benjamin K.

AU - Li, Yingming

AU - Thompson, Melissa G.

AU - Gleason, Joseph

AU - Chen, Zhi

AU - Malaeb, Bahaa

AU - Corey, David R.

AU - Herbert, Brittney Shea

AU - Shay, Jerry W.

AU - Koeneman, Kenneth S.

PY - 2006/1/1

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N2 - Purpose: Determine the efficacy and timing of small molecule oligonucleotide-based inhibitors to the enzyme telomerase in an in vitro model of androgen-independent, osseous prostate cancer. Materials and Methods: Telomerase was inhibited in prostate cancer cell lines C4-2/C4-2B and in controls by using small molecule antisense oligonucleotide-based inhibitors alone or in various combinations of small-dose Taxotere® (sanofi-aventis, Bridgewater, NJ) and/or conditionally replication competent adenovirus (AD-BSP-E1a). After transfection and proliferation, telomerase telomeric repeat amplification protocol and telomere restriction fragment assays were performed, with specific times for evaluating telomere length. Specimens were stained for analysis with hematoxylin and eosin (H&E), terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling (TUNEL), and prostate-specific antigen (PSA). Results: C4-2/C4-2B cell lines had the shortest initial mean telomere length (approximately 2.5 kilobase [kb]) compared to PC-3 (approximately 5.5 kb). Dose-dependent inhibition of telomerase activity was seen using match oligonucleotide-based inhibitors to telomerase (50% inhibitory concentration 3-5 nm), whereas mismatch compound showed no telomerase inhibition. Significant growth delay and apoptosis in cell lines occurred after >50 days of treatment. Cells treated with combination "triple therapy" (i.e., telomerase inhibitors, adenovirus, and Taxotere®) had the highest amount of apoptosis. Compared to controls, all combination treatment groups had statistically significant reductions in prostate-specific antigen in the conditioned media. Conclusions: Combining cytotoxic regimens with small molecule inhibitors to telomerase with oligonucleotide-based agents could be beneficial in controlling osseous hormone refractory prostate cancer, as evidenced by these in vitro, preclinical investigations. Telomerase inhibition needs to move into in vivo models and human studies.

AB - Purpose: Determine the efficacy and timing of small molecule oligonucleotide-based inhibitors to the enzyme telomerase in an in vitro model of androgen-independent, osseous prostate cancer. Materials and Methods: Telomerase was inhibited in prostate cancer cell lines C4-2/C4-2B and in controls by using small molecule antisense oligonucleotide-based inhibitors alone or in various combinations of small-dose Taxotere® (sanofi-aventis, Bridgewater, NJ) and/or conditionally replication competent adenovirus (AD-BSP-E1a). After transfection and proliferation, telomerase telomeric repeat amplification protocol and telomere restriction fragment assays were performed, with specific times for evaluating telomere length. Specimens were stained for analysis with hematoxylin and eosin (H&E), terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling (TUNEL), and prostate-specific antigen (PSA). Results: C4-2/C4-2B cell lines had the shortest initial mean telomere length (approximately 2.5 kilobase [kb]) compared to PC-3 (approximately 5.5 kb). Dose-dependent inhibition of telomerase activity was seen using match oligonucleotide-based inhibitors to telomerase (50% inhibitory concentration 3-5 nm), whereas mismatch compound showed no telomerase inhibition. Significant growth delay and apoptosis in cell lines occurred after >50 days of treatment. Cells treated with combination "triple therapy" (i.e., telomerase inhibitors, adenovirus, and Taxotere®) had the highest amount of apoptosis. Compared to controls, all combination treatment groups had statistically significant reductions in prostate-specific antigen in the conditioned media. Conclusions: Combining cytotoxic regimens with small molecule inhibitors to telomerase with oligonucleotide-based agents could be beneficial in controlling osseous hormone refractory prostate cancer, as evidenced by these in vitro, preclinical investigations. Telomerase inhibition needs to move into in vivo models and human studies.

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