Establishment of over-expressed cell line expressing Clonorchis sinensis CsSBAT

초록

Clonorchiasis by infection of Clonorchis sinensis causes clinical symptoms, such as obstructive jaundice, dull epigastric pain, bile duct epithelial damage, biliary stones, cholangitis, cholecystitis, hepatomegaly, hepatic fibrosis, and cholangiocarcinoma. We have reported the properties of Clonorchis sinensis sodium-dependent bile acid 채- transporter (CsSBAT) as a steroid sulfate conjugate transporter. Here we report on establising of a permanently over-expressed cell line expressing CsSBAT. CsSBAT cDNA was sub-cloned between XhoI and BamHI sites of ipEGFP-C1 vector. The cDNA was prepared by PCR with restriction enzyme sites introduced primer sets. Transfection into the COS-7 cells was performed using TransIT-2020 transfection reagent. After 2 days from transfection, cells were seeded 100 cells/ 10 cm culture dish for determination of fluorescence. Selected cells were determined the transport properties using tritium labeled estrone sulfate. The uptake of estrone sulfate is incubation time- and soedium-dependent manner. From the results,an established cell line will be a good screening system for inhibitors for new drug development.