The Effect Of P2X₇ Receptor On The Expression Of CD44 And Its Function In P388D1 Cells

Objective To study the functional expression of P2X7 receptor in P388D1 and its association with CD44 and cancer cells metastasis.Methods Analysis of P2X7R expression by RNA extraction and revers transcriptasepolymerase chain reaction (RT-PCR),western blotting and analysis of P2X7R function by the bleb formation upon agonist stimulation was observed under microscope. Extracellular adenosine triphosphate(ATP) is P2X7R agonist .Analysis of P388D1 cell CD44 expression after incubated for up to 30 minutes in NaCl or KCl medium at 37°C with ATP by Flow CytometryMoreover,KN-62, the most potent inhibitor for the P2X7R were added 15 minutes before the addition of ATP then analysis of P388D1 cell CD44 expression by Flow Cytometry.Results P2X7 receptor expression could be detected in P388D1 at mRNA and protein level. The result of beble formation suggest the P388D1 cells express functional P2X7 receptors.Extracellular ATP causes fluorescence intensity of CD44 attenuating.KN62 Inhibited ATP-induced causes fluorescence intensity of CD44 attenuating by P2X7R. trypan blue staining results of the less number of the cells in bottom chamber which were stained with trypan blue showed that the cells passed through the filter and attached to the bottom chamber. The number of P388D1 cells was decreased compared without the P2X7R actived by ATP.Conclusion Base on our study, it is indicated that P2X7R can descrease expression of CD44 in P388D1.It will open a new vision for the clinic treatment of tumor.