Impact of CRISPR/Cas9-Mediated CD73 Knockout in Pancreatic Cancer

• Published in: Cancers Vol. 15; no. 19; p. 4842
• Main Authors: Zhang, Jinping, Zhang, Shuman, Dörflein, Isabella, Ren, Xiaofan, Pfeffer, Susanne, Britzen-Laurent, Nathalie, Grützmann, Robert, Duan, Xianglong, Pilarsky, Christian
• Format: Journal Article
• Language: English
• Published: Basel MDPI AG 01.10.2023; MDPI
• Subjects: Bioinformatics; Cancer therapies; CD73; CD73 antigen; Cell cycle; Cell migration; Cell proliferation; Cells; CRISPR; CRISPR/Cas9; deep sequencing; E-cadherin; Ethylenediaminetetraacetic acid; Extracellular signal-regulated kinase; Flow cytometry; Genes; Germany; Health aspects; Immune system; Immunotherapy; Kinases; Medical prognosis; Medical research; Mortality; Pancreatic cancer; Plasmids; Scientific equipment and supplies industry; Stat3 protein; Tumor cells; United States; United States; Germany; United States > US
• ISSN: 2072-6694; 2072-6694
• DOI: 10.3390/cancers15194842

Pancreatic cancer is among the cancers with the highest mortality rates. Most of the patients are found to have advanced cancer, losing the chance of surgical treatment, and there is an urgent need to find new treatment methods. Targeted therapy for specific genes that play a key role in cancer is now an important means to improve the survival rate of patients. We determined that CD73 is highly expressed in pancreatic cancer by flow cytometry and qRT-PCR assays combined with bioinformatics techniques. Application of CRISPR/Cas9 technology to knockout CD73 in human and murine cell lines, respectively, revealed that CD73 inactivation inhibited cell growth and migration and induced G1 cell cycle arrest. We also found that CD73 deletion inhibited the ERK/STAT3 pathway and activated the E-cadherin pathway. In addition, a CRISPR/Cas9 protein kinase library screen was performed and identified Pbk, Fastk, Cdk19, Adck5, Trim28, and Pfkp as possible genes regulating CD73.