Construction and Feasibility Analysis of an EPC1 Overexpression Cell Model of Hepatic Alveolar Echinococcosis in Vitro

Abstract

Objective: To explore the feasibility of establishing a cell model with high expression of Echinococcus protoscolex calcium binding protein 1 (EPC1) in vitro. Methods: The target gene EPC1 (AF481884) information was synthesised and connected to the expression vector pcDNA3.4. The constructed positive plasmid was named EPC1-pcDNA3.4. The constructed overexpression plasmid vector was transfected into six groups, A1, A2, B1, B2, C1 and C2, after ultrapure extraction to remove the endotoxin. Each group contained three replicates, with A1, B1 and C1 as control groups and A2, B2 and C2 as experimental groups. Group A1 consisted of human lung cancer cells H1299. Group A2 consisted of human lung cancer cells H1299 + EPC1 vector transfection. Group B1 contained human liver cancer cells, HepG2. Group B2 comprised human liver cancer cells HepG2 + EPC1 vector. Group C1 consisted of normal hepatocyte HL7702. Group C2 contained normal hepatocyte HL7702 + EPC1 vector. The messenger ribonucleic acid (mRNA) and protein expression of EPC1 in groups A1, A2, B1, B2, C1 and C2 were determined by quantitative real-time PCR (qPCR) and western blotting, respectively. Results: The qPCR results showed that EPC1 mRNA was detected in groups A2, B2 and C2 but not in groups A1, B1 and C1. Western blotting results showed that EPC1 protein level was not expressed in any group. Conclusions: The high-expression cell model of EPC1 in hepatic alveolar echinococcosis was constructed in vitro, which is of great significance for diagnosing hepatic echinococcosis. The EPC1 protein is not expressed in vitro, which may be related to the expression vector and inflammatory response.