Cellular and Molecular Research Center, Research Institute for Health Development, Kurdistan University of Medical Sciences, Sanandaj, Iran.
Cancer and Immunology Research Center, Research Institute for Health Development, Kurdistan University of Medical Sciences, Sanandaj, Iran.
Gulf J Oncolog. 2023 Jan;1(41):7-16.
Prostate cancer is the most common malignant cancer among men and is the second deadliest cancer in men after lung cancer. Understanding the molecular mechanisms involved in development and progression of prostate cancer is essential to improve both diagnostic and therapeutic strategies in this regard. In addition, using novel gene therapy-based methods for treatment of cancers has gotten increasing attention during the recent years. Accordingly, this study was aimed to evaluate the inhibitory effect of MAGE-A11 gene, as an important oncogene involved in the pathophysiology of prostate cancer invitro model. The study was also aimed to evaluate the downstream genes related to MAGE-A11.
First, MAGE-A11 gene was knocked out in PC-3 cell line using "Clustered regularly interspaced short palindromic repeats" (CRISPR)/ "CRISPR-associated genes 9" (CRISPR/Cas9) method. Next, the expression levels of MAGE-A11, survivin and Ribonucleotide Reductase Small Subunit M2 (RRM2) genes were determined by quantitative polymerase chain reaction (qPCR) technique. The levels of proliferation and apoptosis were also analyzed in PC-3 cells using CCK-8 and Annexin V-PE/7-AAD assays.
The results showed that the disruption of MAGE-A11 by CRISPR/Cas9 method significantly decreased proliferation (P< 0.0001) and enhanced apoptosis (P< 0.05) in PC-3 cells compared to control group. Moreover, the disruption of MAGE-A11 significantly down regulated the expression levels of survivin and RRM2 genes (P< 0.05).
Our results demonstrated that knocking out MAGE-11 gene by CRISPR/CAS9 technique could efficiently inhibit cell proliferation and induce apoptosis in PC3 cells. Survivin and RRM2 genes might also participated in these processes.
前列腺癌是男性中最常见的恶性肿瘤，也是仅次于肺癌的男性第二大癌症杀手。了解前列腺癌发生和发展过程中的分子机制对于改善这方面的诊断和治疗策略至关重要。此外，近年来，使用基于新型基因治疗的方法治疗癌症受到了越来越多的关注。因此，本研究旨在评估 MAGE-A11 基因作为参与前列腺癌病理生理学的重要癌基因在体外模型中的抑制作用。本研究还旨在评估与 MAGE-A11 相关的下游基因。
首先，使用“成簇规律间隔短回文重复”（CRISPR）/“CRISPR 相关基因 9”（CRISPR/Cas9）方法敲除 PC-3 细胞系中的 MAGE-A11 基因。接下来，通过定量聚合酶链反应（qPCR）技术测定 MAGE-A11、survivin 和核糖核苷酸还原酶小亚基 M2（RRM2）基因的表达水平。还使用 CCK-8 和 Annexin V-PE/7-AAD 测定法分析 PC-3 细胞中的增殖和凋亡水平。
结果表明，与对照组相比，CRISPR/Cas9 方法敲除 MAGE-A11 基因显著降低了 PC-3 细胞的增殖（P<0.0001）并增强了凋亡（P<0.05）。此外，敲除 MAGE-A11 基因显著下调 survivin 和 RRM2 基因的表达水平（P<0.05）。
我们的结果表明，CRISPR/CAS9 技术敲除 MAGE-11 基因可有效抑制 PC3 细胞的增殖并诱导其凋亡。Survivin 和 RRM2 基因也可能参与这些过程。
