Open Access Research Article Article ID: SJGGT-1-101

    In LNCaP Cells Inhibition of BCL-2 by Antisense Oligonucleonucleotides Results in Compensatory Changes in Apoptosis

    Marvin Rubenstein*, Courtney M.P. Hollowell and Patrick Guinan

    Antisense oligonucleotides (oligos) have been evaluated for treating prostate cancer in both in vivo and in vitro models. Although most oligos contain a single mRNA binding site, our laboratory evaluates bi-specific oligos directed towards two proteins. This study evaluates the growth inhibition in vitro of the LNCaP cell line employing mono- and bispecific oligos directed against BCL-2 [the second binding site was directed against the epidermal growth factor receptor (EGFR)]. These oligos were administered with lipofectin as part of a nanoparticle delivery system. Additionally, g, the expression of five apoptosis regulatory proteins, BCL-2, bax, caspase-3, clusterin and AKT-1 was evaluated by RTPCR.

    LNCaP prostate tumor cells were incubated in the presence of oligos specifically directed against BCL-2 and their effect was compared to lipofectin containing controls. Significant, but comparable, growth inhibition was produced by both mono- and bi-specific forms. Employing RT-PCR to determine BCL-2 expression, we found that the greatest amount of mRNA suppression approached 100% for each type of oligo with mono-specific MR4 (directed only against BCL-2) equal 100%; and bispecifics MR24 and MR42, being 86% and 100% respectively. Based upon both inhibition of cell growth and BCL-2 expression, bi-specific antisense oligos directed against EGFR and BCL-2 mRNAs are at least as effective as a mono-specific directed solely towards BCL-2.

    In an effort to determine a compensatory response by cells needed to evade apoptosis in the presence of BCL-2 suppression, the levels of mRNA encoding non-targeted bax, caspase-3, clusterin and AKT-1 were evaluated. Suppression of the apoptosis inhibitor (BCL-2) in LNCaP cells did not alter either bax or clusterin expression. However, the expression of non-targeted caspase-3 (an apoptosis promoter) was suppressed and the expression of non-targeted AKT-1 (an apoptosis inhibitor) was enhanced. This suggests that tumor variants can resist apoptosis through the altered expression of non-targeted regulators of apoptosis. If BCL-2 suppression was to be clinically targeted by antisense oligos, similar experiments may be helpful in identifying genes with a similar function.

    Keywords: Antisense oligonucleotides; Prostate cancer; BCL-2; bax; caspase-3; clusterin; AKT-1; Therapy

    Published on: Mar 27, 2014 Pages: 1-6

    Full Text PDF Full Text HTML DOI: 10.17352/sjggt.000001
    CrossMark Publons Harvard Library HOLLIS Search IT Semantic Scholar Get Citation Base Search Scilit OAI-PMH ResearchGate Academic Microsoft GrowKudos Universite de Paris UW Libraries SJSU King Library SJSU King Library NUS Library McGill DET KGL BIBLiOTEK JCU Discovery Universidad De Lima WorldCat VU on WorldCat


    Case Reports

    Pinterest on SJGGT

    Help ? Google Reviews 11