title: [Establishment of HEK293 cell lines stably expressing human parathyroid hormone receptors].
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 constructs 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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Department of Spinal Surgery, Southern Medical University, Guangzhou, China. mengspine@163.com
authors.forename: Yue
authors.initials: Y
authors.lastname: Meng
authors.forename: Miaomiao
authors.initials: M
authors.lastname: Xie
authors.forename: Zhen
authors.initials: Z
authors.lastname: Lin
authors.forename: Liang
authors.initials: L
authors.lastname: Yuan
authors.forename: Wei
authors.initials: W
authors.lastname: Li
authors.forename: Song
authors.initials: S
authors.lastname: Hao
authors.forename: Dehong
authors.initials: D
authors.lastname: Yang
mesh_terms: D015870:Gene Expression; D005822:Genetic Vectors; D057809:HEK293 Cells; D006801:Humans; D010957:Plasmids; D018016:Receptors, Parathyroid Hormone / Q000235:genetics* / Q000378:metabolism; D017422:Sequence Analysis, DNA; D015398:Signal Transduction / Q000235:genetics; D014162:Transfection
chemical_list: D018016:Receptors, Parathyroid Hormone