Laboratory of Medical Investigation (LIM55), Urology Department, University of Sao Paulo Medical School, Sao Paulo, Brazil.
BMC Urol. 2012 May 29;12:14. doi: 10.1186/1471-2490-12-14.
Prognosis of prostate cancer (PCa) is based mainly in histological aspects together with PSA serum levels that not always reflect the real aggressive potential of the neoplasia. The micro RNA (miRNA) mir-21 has been shown to regulate invasiveness in cancer through translational repression of the Metaloproteinase (MMP) inhibitor RECK. Our aim is to investigate the levels of expression of RECK and miR-21 in PCa comparing with classical prognostic factors and disease outcome and also test if RECK is a target of miR-21 in in vitro study using PCa cell line.
To determine if RECK is a target of miR-21 in prostate cancer we performed an in vitro assay with PCa cell line DU-145 transfected with pre-miR-21 and anti-miR-21. To determine miR-21 and RECK expression levels in PCa samples we performed quantitative real-time polymerase chain reaction (qRT-PCR).
The in vitro assays showed a decrease in expression levels of RECK after transfection with pre-miR-21, and an increase of MMP9 that is regulated by RECK compared to PCa cells treated with anti-miR-21. We defined three profiles to compare the prognostic factors. The first was characterized by miR-21 and RECK underexpression (N = 25) the second was characterized by miR-21 overexpression and RECK underexpression (N = 12), and the third was characterized by miR-21 underexpression and RECK overexpression (N = 16). From men who presented the second profile (miR-21 overexpression and RECK underexpression) 91.7% were staged pT3. For the other two groups 48.0%, and 46.7% of patients were staged pT3 (p = 0.025).
Our results demonstrate RECK as a target of miR-21. We believe that miR-21 may be important in PCa progression through its regulation of RECK, a known regulator of tumor cell invasion.
前列腺癌 (PCa) 的预后主要基于组织学方面以及 PSA 血清水平,而 PSA 血清水平并不总是反映肿瘤的真实侵袭潜能。微 RNA (miRNA) mir-21 已被证明通过金属蛋白酶 (MMP) 抑制剂 RECK 的翻译抑制来调节癌症的侵袭性。我们的目的是通过比较经典的预后因素和疾病结果来研究 RECK 和 miR-21 在 PCa 中的表达水平,并在体外使用 PCa 细胞系进行试验以检测 RECK 是否是 miR-21 的靶标。
为了确定 RECK 是否是前列腺癌细胞中的 miR-21 的靶标,我们使用转染了 pre-miR-21 和 anti-miR-21 的 PCa 细胞系 DU-145 进行了体外试验。为了确定 PCa 样本中的 miR-21 和 RECK 表达水平,我们进行了定量实时聚合酶链反应 (qRT-PCR)。
体外试验显示,转染 pre-miR-21 后 RECK 的表达水平下降,与用 anti-miR-21 处理的 PCa 细胞相比,RECK 调节的 MMP9 增加。我们定义了三种谱来比较预后因素。第一个谱的特征是 miR-21 和 RECK 表达下调(N=25),第二个谱的特征是 miR-21 过表达和 RECK 表达下调(N=12),第三个谱的特征是 miR-21 表达下调和 RECK 过表达(N=16)。在表现出第二种谱(miR-21 过表达和 RECK 表达下调)的男性中,91.7%的患者分期为 pT3。对于其他两组,48.0%和 46.7%的患者分期为 pT3(p=0.025)。
我们的结果表明 RECK 是 miR-21 的靶标。我们认为,miR-21 可能通过其对 RECK 的调节在 PCa 进展中发挥重要作用,RECK 是肿瘤细胞侵袭的已知调节剂。