PRSS3 promotes immune escape of non-small cell lung cancer by activating JAK2/STAT3 signaling pathway and upregulating PD-L1 expression via FGFR2
Objective To investigate the role of PRSS3/FGFR2 in immune escape of non-small cell lung cancer(NCLC)cells by using an A549 cell model which stably knocks down PRSS3 and overexpresses FGFR2 on the basis of PRSS3.Methods A549 cells were divided into sh-NC group,sh-PRSS3 group,sh-PRSS3+ov-NC group and sh-PRSS3+ov-FGFR2 group,and were transfected with corresponding lentiviruses respectively.Western blot was used to detect the expression of PRSS3,FGFR2,p-JAK2,JAK2,p-STAT3,STAT3 and PD-L1 in A549 cells of different groups;CCK-8 was used to detect the proliferation of theses A549 cells;Transwell was used to detect the migration and invasion ability of theses A549 cells.Furhtermore,CD8+T cells were co-cultured with these A549 cells,and trypan blue staining and flow cytometry were used to detect the viability and apoptosis rate of the CD8+T cells,respectively.The levels of sPD-L1,IFN-γ,TNF-α,granzyme B and perforin in the supernatant of co-cultured cells were detected by ELISA.Results Compared with sh-NC group,sh-PRSS3 group and sh-PRSS3+ov-NC group demonstrated lower expressions of PRSS3,FGFR2,p-JAK2,p-STAT3 and PD-L1 proteins,as well as lower levels of proliferation activity,the number of migrating cells and the number of invading cells of A549 cells.Furthermore,in the supernatant of co-culture medium of sh-PRSS3 and sh-PRSS3+ov-NC groups,the viability of CD8+T cells was increased,the levels of IFN-γ,TNF-α,granzyme B and perforin were elevated,while the apoptosis of CD8+T cells and the level of sPD-L1 were decreased,as compared with sh-NC group.In the sh-PRSS3+ov-FGFR2 group,all these differences mentoned above were mitigated,which means the effects of sh-PRSS3 were antagonized by overexpression of FGFR2.Conclusion PRSS3 induces immune escape of NSCLC cells by up-regulating FGFR2 expression,and its mechanism may be related to the up-regulation of PD-L1 expression induced by activating JAK2/STAT3 signaling pathway.
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维普
