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

SIMPLE SUMMARY: Pancreatic cancer is one of the cancers with the highest mortality rate, and targeted therapy against specific genes that play a crucial role in cancer is the latest therapeutic approach. This study showed that CD73 is highly expressed in pancreatic cancer, and the application of CRISPR/Cas9 technology to knockout of CD73 in human and murine cell lines can inhibit the proliferation and migration of tumor cells and induced G1 cell cycle arrest. We also found that CD73 deletion inhibited the ERK/STAT3 pathway and activated the E-cadherin pathway. Pbk, Fastk, Cdk19, Adck5, Trim28, and Pfkp are the genes that may regulate CD73 in pancreatic cancer. ABSTRACT: 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.