SAHA Up-regulates HLA-? Expression By Activating The STAT1 Signaling Pathway In Non-small Cell Lung Cancer

Objective:To investigate the mechanisms of histone deacetylase inhibitor(HDACI): vorinostat / Suberoylanilide hydroxamic acid(SAHA)up-regulated expression of human lymphocyte antigen class I(HLA-?)in non-small cell lung cancer.Methods:(1)CCK8 experiment was used to determine the optimal drug concentration of SAHA in A549 and H520 cells(2)q RT-PCR,Western blot and flow cytometry respectively were used to detect the expression of HLA-? after treatment with SAHA in both A549 and H520 cells;(3)Western blot detect the expression of STAT1 after treatment with SAHA,To investigate the expression of phosphorylated STAT1(Phospho-STAT1,P-STAT1)after SAHA action,we set a time-dose gradient.At the same time,nuclear-plasma separation experiments were used to detect the change of STAT1 nucleus expression after SAHA treatment;(4)si RNA experiment was used to detect the change of HLA-? expression after interference with STAT1,and the expression of HLA-? was detected after adding STAT1 pathway inhibitor.Results:(1)CCK8 experiment showed that the optimal concentration of SAHA was 3umol/L in A549 and H520 cell lines;(2)q RT-PCR results showed that compared with the control group,the expression of HLA-? was up-regulated at the gene level after treatment with SAHA;Western blot results showed that compared with the control group,the expression of HLA-? was up-regulated at the protein level after treatment with SAHA;Flow cytometry showed that compared with the control group,the expression of HLA-? on the cell surface was enhanced after treatment with SAHA,the results were all statistically significant,P < 0.05;(3)Western blot results showed that SAHA can up-regulate STAT1 expression and increase the expression of P-STAT1.Nuclear separation studies showed that SAHA can promote STAT1 into the nucleus;(4)Western blot results showed that the expression of HLA-? was decreased after si RNA-STAT1 transfection compared with si RNA-NC transfection control group.Conclusion:SAHA up-regulates the expression of HLA-? by activating the STAT1 signaling pathway in non-small cell lung cancer,It is speculated that SAHA can promote the immune response process of non-small cell lung cancer in this way,which providing a new direction for the treatment of non-small cell lung cancer by SAHA.