Abstract: Objective To establish HEK293 cell lines with stable expression of human parathyroid hormone (PTH) receptors.
Methods The purified gene fragments of PTH-related peptide receptor (PTHR) and its mutant form (DSEL) were cloned
separately into pcDNA3.1(+) vector after digestion with EcoR I and Not I, and the resulted pcDNA3.1(+)-PTHR and pcDNA3.1
(+)-DSEL plasmids were verified by restriction enzyme digestion and DNA sequencing. HEK293 cells were transfected with
these plasmids and the expression of PTHR and DSEL in the cells were examined by RT-PCR and ELSIA. Results Sequencing
and restriction enzyme digestion analysis showed that PTHR and DSEL cDNAs were correctly cloned into pcDNA3.1(+)vector.
After a 48-h transfection of HEK293 cells with the recombinant plasmids and G418 selection, the positive cell clones stably
expressing the constucts were obtained, which showed expressions of PTHR and DSEL mRNAs detected by RT-PCR. These
positive cells showed high levels of PLC and aAMP production in response to PTH stimulation. Conclusion The HEK293 cell
lines with stable expression of PTH1R or DSEL gene established in this study provide useful cell models for studying the
physiological functions of PTH peptides.