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Published on: April 27, 2018
The Disruption of Mage-11 Gene via CRISPR/Cas9 Method Induced Apoptosis in the in vitro Model of Prostate Cancer
Leila Farhadi1,2, Farzad Soleimani1, Shohreh Fakhari2
1Cellular and Molecular Research Center, Research Institute for Health Development, Kurdistan University of Medical Sciences, Sanandaj, Iran.
Backgrounds And Aims:
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.
Materials And Methods:
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.
Results:
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).
Conclusion:
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.
Insights
Knocking out the MAGE-A11 oncogene using CRISPR/Cas9 significantly reduced prostate cancer cell proliferation and increased apoptosis. This disruption also downregulated survivin and RRM2 gene expression, suggesting their role in prostate cancer progression.
Area of Science:
- Oncology
- Molecular Biology
- Gene Therapy
Background:
- Prostate cancer is a leading cause of cancer death in men, necessitating improved diagnostic and therapeutic strategies.
- Understanding the molecular underpinnings of prostate cancer is crucial for developing effective treatments.
- Gene therapy presents a promising avenue for novel cancer treatment approaches.
Purpose of the Study:
- To investigate the inhibitory effects of MAGE-A11 gene knockout on prostate cancer cells in vitro.
- To assess the impact of MAGE-A11 disruption on cell proliferation and apoptosis.
- To explore the downstream gene expression changes associated with MAGE-A11 inhibition.
Main Methods:
- Utilized the CRISPR/Cas9 system for targeted knockout of the MAGE-A11 gene in the PC-3 prostate cancer cell line.
- Quantified gene expression levels of MAGE-A11, survivin, and RRM2 using quantitative polymerase chain reaction (qPCR).
- Assessed cell proliferation and apoptosis using CCK-8 and Annexin V-PE/7-AAD assays, respectively.
Main Results:
- CRISPR/Cas9-mediated MAGE-A11 knockout significantly decreased PC-3 cell proliferation (P<0.0001).
- MAGE-A11 disruption led to a significant enhancement of apoptosis in PC-3 cells (P<0.05).
- The knockout of MAGE-A11 resulted in significant downregulation of survivin and RRM2 gene expression (P<0.05).
Conclusions:
- MAGE-A11 gene knockout via CRISPR/Cas9 effectively inhibits prostate cancer cell proliferation and induces apoptosis.
- Survivin and RRM2 gene expression are potentially involved in the MAGE-A11-mediated regulation of prostate cancer cell behavior.
