The Effect Of Sodium Valpproate In γδT Cells On Lung Cancer A549Line Cells

Objiective: To research the effects of histone deacetylase inhibitorSodium Valproate (HDACi-VPA) on the protein MICA of lung cancerA549lines cells, and also to study the effect of VPA in the cytotoxic effectsmediated by NKG2D-MICA signaling pathway of peripheral blood γδTcells on lung cancer A549lines cells.Methods: Peripheral blood mononuclear cells (PBMC) were collectedfrom health human blood with the ficoll density gradient centrifugationmethods, and we got the γδT cells from the PBMC with Flow cytometryafter3days cultivated, then the Interleukin-2(IL-2,200IU/ml) and thebisphosphonate Zoledronic acid(ZA,300pg/ml) were used to stimulate andexpand the γδT cells; meanwhile the lung cancer A549lines cells werecultured with different concentration of VPA, after24h cultivated, wewould detected the change of MICA protein with the Western blot andPCR methods, and getting the best concentration of VPA. The purificationγδT cells and exponential phase A549lines cells were co-cultured withdifferent E:T(effector:target), then the cytotoxicity of γδT cells againstA549cells was tested with LDH method after24hours cultured, and at the same time the best we would get the best E:T rate; the γδT cells and lungcancer A549line cells were co-cultured with best E:T rate, and the VPAand MICA-Ab(best concentration) were used to change the effect of γδTcells on lung cancer A549line cells, then the cytotoxicity of γδT cellsagainst A549cells was tested after24hours cultured, meanwhile theexpression of MICA and the change of cellular morphology of lung cancerA549observed by immune fluorescence confocal.Results: The results of PCR and Western blot showed that the VPAcould enhance the expression of MICA，and the expression of MICAincreased distinctly with1.0mmol/L concentration of VPA; The results ofLHD showed that the purification γδT cells stimulated and expanded byIL-2and ZA in vitro showed strong cytotoxic effects towards lung cancerA549lines cells when the E:T rate was20:1(the apoptosis inhibition ratewas0.46±0.04), and the cytotoxicity of group(the apoptosis inhibition ratewas0.64±0.05) with VPA was stronger than the group(the apoptosisinhibition rate was0.46±0.04) without VPA(P<0.05), and the cytotoxicityof the group (the apoptosis inhibition rate was0.19±0.05) with MICA-Abdescend distinctly compared with the goup(the apoptosis inhibition ratewas0.46±0.04) without MICA-Ab(P<0.05); The results of immunefluorescence confocal showed that the MICA protein was on the membrane,and the completeness of the cell membrane was destroyed, and the completeness of membrane from the group with MICA-Ab was better thenthat from the group without MICA-Ab.Conclusion: γδT cells stimulated and expanded in vitro showedstrong cytotoxic on lung cancer A549lines cells; the histone deacetylaseinhibitor VPA can enhance the expression of MICA, and also increase thecytotoxic of γδT cells on lung cancer A549lines cells. At the same time,the research shows that γδT combined with VPA is feasible for the cellularimmunity therapy for the lung cancer.