Expression Characteristic Of The Equilibrative Nucleoside Transporter 1 In Human Hepatocellular Carcinoma Cells

Objective:To investigate the expression characteristic of ENT1 in different human hepatoma cell lines and different cell cycles.Methods:1.Human hepatoma cell lines HuH-7,HepG2 and Hep3B, the breast cancer cell lines MCF7 was chose as control group, the localized expression of ENT1 was detected by immunofluorescence staining.2.The expression of ENT1mRNA in human hepatoma cell lines and the breast cancer cell lines was detected by reverse transcriptionpolymerase chain reaction (RT-PCR).3.Cells were devided into 2 groups (Control group:treatmented with Roscovitine 0μmol/L for 24h, Experimental group:treatmented with Roscovitine 32μmol/L for 24h),flow cytometry was used to detected the cell cycle distribution.4.After CDKs were interfered, the expression characteristic of ENT1mRNA was detected by RT-PCR.Results:1.The result of immunofluorescence staining showed that the expression of ENT1 could be seen on three hepatocellular carcinoma cell membranes.2.RT-PCR showed that the expression level of ENT1mRNA in hepatoma cell lines was higher than the breast cancer cell lines',the expression of ENT1mRNA was different among different hepatoma cell lines.3.Flow cytometry showed that the ratio of HuH-7 cells in Go/Gi phase was increased(76.29±0.34%vs 58.50±0.33%, P<0.05)with Roscovitine 32μmol/L treatmented for 24h, in S phase was increased (25.55±0.15%vsl5.19±0.34%, P<0.05),in G2/M phase was increased (16.35±0.31%vs8.51±0.41%,P<0.05).4.The expression level of ENT1mRNA was increased after HuH-7 cells were blocked in G0/G1 phase. Conclusions:1.The expression of ENT1 is common on hepatocellular carcinoma cell membrane.2.The expression level of ENT1mRNA in hepatoma cell lines is higher than the breast cancer cell lines',the expression of ENT1mRNA is different among different hepatoma cell lines.3.At the G0/G1 phase of the cell, the expression level of ENT1mRNA is significantly increased.