The biological effects of HPT protein on the drug resistance of gastric cancer cell / 解放军医学杂志

ABSTRACT

Objective To prepare an antiserum of human putative magnesium transporter protein (HPT protein) to study its biological effects on the drug resistance of gastric cancer cell by observing its expression level. Methods The cDNA encoding HPT protein was cloned and sequenced. Prokaryotic expression vector was constructed and HPT protein fused with histidine tag was expressed. Mice were immunized with the polyacrylamide gel particles containing the fusion protein, and the antisera of them were detected for antibody activity by Western blot. Using the antiserum, HPT protein expression level was assessed in different gastric cancer cell line. Results The sequence of cloned cDNA fragment was consistent with that of HPT. After BL21/pRSETB bacterium was induced with IPTG, a new protein band with a relative molecular weight of 55kD was showed on SDS PAGE profile, and further proved to be fusion protein with histidine tag by Western blot. The antiserum of immunized mice only recognized a unique protein band with a relative molecular weight of 50kD. HPT protein expression level in SGC7901/ADR was higher than in SGC7901. Conclusion The antiserum of HPT protein was successfully prepared, and HPT was found to probably take part in the gastric cancer cell MDR