Division of Cellular Biology, Hektoen Institute for Medical Research, 2240 West Ogden Avenue, Chicago, IL 60612, USA.
In Vivo. 2011 Jan-Feb;25(1):61-7.
Antisense oligonucleotides (oligos) have been employed against in vivo and in vitro prostate cancer models. Most oligos consist of a single mRNA binding site, targeting a single gene product or others sharing sequence homology. However, our lab has developed bispecifics directed towards two (including unrelated) proteins. Previously we have shown that mono- and bispecific oligos targeting BCL-2 significantly inhibit LNCaP cell growth. Employing reverse transcriptase-polymerase chain reaction we found comparable suppression of expressed BCL-2. Computer models suggested that this activity could, in part, be enhanced by the formation of siRNA-like double-stranded regions, generated by intrastrand base pair complementarity. We hypothesize that these regions could be interferon inducers (like poly I:C) and enhance the expression of prostate specific cell surface antigens. The expression of cell surface prostate-specific membrane antigen (PSMA) and the secreted prostate-specific antigen (PSA) were candidates for evaluation. To test this theory, we evaluated the effects of mono- and bispecific oligos (with intrastrand complementarity), targeting BCL-2, upon the expression of non-targeted proteins PSMA, PSA and interferon-gamma (IFN-γ) in LNCaP cells. Levels of mRNA encoding PSMA were significantly elevated following treatment with the bispecific oligos (directed against both BCL-2 and the epidermal growth factor receptor) but not by the monospecific directed solely against BCL-2. Furthermore, no differences were detected in mRNA levels encoding PSA following treatment with either mono- or bispecific forms. IFN-γ expression was also significantly increased by the bispecific and not by the monospecific oligos, supporting the hypothesis of interferon induction. This suggests that prostate cells (including LNCaP) retain an endogenous interferon-based antiviral defense mechanism (similar to that found in the testes) which is induced by double stranded oligos. Enhanced expression of cell surface differentiation antigens (such as PSMA) could increase targeting by cytotoxic T-cells and potentiate prostate cancer vaccines directed against tumor-associated cell surface antigens.
反义寡核苷酸（oligos）已被用于体内和体外前列腺癌模型。大多数 oligos 由一个单个的 mRNA 结合位点组成，靶向一个单一的基因产物或其他具有序列同源性的基因产物。然而，我们的实验室已经开发了针对两个（包括不相关的）蛋白质的双特异性试剂。以前，我们已经表明，靶向 BCL-2 的单和双特异性 oligos 显著抑制 LNCaP 细胞的生长。通过逆转录聚合酶链反应，我们发现表达的 BCL-2 受到类似的抑制。计算机模型表明，这种活性部分可以通过由链内碱基互补产生的 siRNA 样双链区域来增强。我们假设这些区域可能是干扰素诱导剂（如聚 I:C），并增强前列腺特异性细胞表面抗原的表达。前列腺特异性膜抗原（PSMA）和分泌的前列腺特异性抗原（PSA）的表达是评估的候选物。为了验证这一理论，我们评估了针对 BCL-2 的单和双特异性 oligos（具有链内互补性）对 LNCaP 细胞中非靶向蛋白 PSMA、PSA 和干扰素-γ（IFN-γ）表达的影响。在用针对 BCL-2 和表皮生长因子受体的双特异性 oligos 处理后，编码 PSMA 的 mRNA 水平显著升高，但在用仅针对 BCL-2 的单特异性 oligos 处理后没有升高。此外，在用单或双特异性形式处理后，编码 PSA 的 mRNA 水平没有差异。IFN-γ 的表达也被双特异性 oligos 显著增加，而不是单特异性 oligos，支持了干扰素诱导的假说。这表明前列腺细胞（包括 LNCaP）保留了一种内源性的基于干扰素的抗病毒防御机制（类似于在睾丸中发现的机制），该机制被双链 oligos 诱导。细胞表面分化抗原（如 PSMA）的表达增强可以增加细胞毒性 T 细胞的靶向性，并增强针对肿瘤相关细胞表面抗原的前列腺癌疫苗。
