The Effect And Molecular Immune Mechanism Of Tanshinone ?A Enhancing NK Cells-medicated Killing Of Non-small Cell Lung Cancer

?Objective?This study aimed to investigate the molecular immune mechanism of Tanshinone ?A,an effective compound of traditional Chinese medicine,improving the effect of NK cells-medicated killing of non-small cell lung cancer cells.?Method?MTT and CCK8 cell activity assay was used to detect the direct inhibitory effect of Tanshinone ?A on non-small cell lung cancer cell lines and NK cells;CD107?and CFSE/7AAD double staining by flow cytometry were used to detect the effect of Tanshinone ?A on the degranulation and special lysis of NK cells against non-small Cell Lung Cancer cells A549,H1299 and H460;the effect of Tanshinone ?A on NK cells that killed A549-luciferase cells by luciferase assay;in vivo,LLC subcutaneous tumor-bearing mice were detect the direct anti-tumor effect of Tanshinone ?A;H460 subcutaneous tumor-bearing model was used to investigate the combined antitumor effect of Tanshinone ?A and NK cells;LLC subcutaneous tumor-bearing mice were used to detect the anti-tumor effect of Tanshinone ?A with NK cell blocking antibody PK136;the ligands on tumor cells were invesigated by flow cytometry after Tanshinone ?A treatments;the lysis of NK cells against A549 and H460 after Tanshinone ?A treated with TRAIL and NKG2 D inhibitor by flow cytometry;Real Time PCR and Western Blot to detect the effect of Tanshinone ?A on PERK/ATF4/CHOP signaling pathway.?Results?1.Tanshinone ?A can directly inhibit the growth of non-small cell lung cancer A549,H1299 and H460 cells;2.The effect of Tanshinone ?A on NK cells growth is not significant;3.Tanshinone ?A can enhance the degranulation effect of NK cells against A549 and H460 cells,but not in H1299 cells;4.Tanshinone ?A can enhance the special lysis of NK cells on A549,H460 and H1299 cells;5.Tanshinone ?A can enhance the killing function of NK cells on A549-luciferase cells;Tanshinone ?A directly inhibits the growth of LLC tumor growth in vivo;7.Tanshinone ?A can be combined with NK cells to enhanced the inhibition effect in H460 tumor-bearing mice;8.after NK cell depletion,the tumor inhibition effect of Tanshinone ?A in LLC subcutaneous tumor was weakened;9.Tanshinone ?A up-regulated DR5 and ULBP1 on A549,H460 and H1299 cells;10.the enhancement of NK cell activity against A549 and H460 by Tanshinone ?A was inhibited by blocking TRAIL and NKG2D;11.Tanshinone ?A activeted PERK/ATF4/CHOP signaling pathway..?Conclusion?1.Tanshinone ?A can enhance the killing activity of NK cells against non-small cell lung cancer A549,H1299,H460;2.Tanshinone ?A can also enhance NK cell activity in vivo and its enhancement effect was inhibit after NK cells depletion;3.Tanshinone ?A could up-regulation of DR5 and ULBP1 expression on the surface of non-small cell lung cancer cells;4.Enhancement of Tanshinone ?A on NK cells activity was inhibited after TRAIL and NKG2 D blockade;5.Up-regulation of DR5 and ULBP1 may be related to activate PERK/ATF4/CHOP signaling pathway..