Bridging intergrator-1 inhibits the expression of PD-L1 through c-MYC pathway in non-small cell lung cancerA549 cells
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Abstract:
Objective: To investigate the effect and mechanism of bridging intergrator-1 (BIN1) on expression of programmed death-ligand 1 (PD-L1) in non-small cell lung cancer (NSCLC) A549 cells. Methods: Quantitative real-time polymerase chain reaction (qRTPCR) and Western blotting were used to detect the mRNA and protein expression of BIN1 and PD-L1 in A549 cells and normal human embryo lung fibroblast 2BS cells, respectively. Eukaryotic expression plasmid CMV-MCS-GFP-SV40-Neomycin-BIN1 containing human full length BIN1 gene sequence was transfected into A549 cells via cationic liposomes by using gene transfection technology (as BIN1+group); c-MYC-siRNA was used to knockdown the expression of c-MYC in A549 cells through RNA interference technique (as c-MYC-siRNA group). The transfection efficiencies were verified by qRT-PCR and Western blotting, the effects of BIN1 over-expression and c-MYC knock-down on the expression of c-MYC and PD-L1 in A549 cells were detected as well. Results: Comparing with 2BS cells, the expression of BIN1 was down-regulated in A549 cells at both mRNA and protein levels, while the expression of PD-L1 was up-regulated (all P<0.05). The expression of BIN1 was increased at both mRNA and protein level in BIN1+ group, while the expression of PD-L1 was decreased significantly after B1N1 transfection (all P<0.05). After transfection of c-MYC-siRNA into A549 cells, the expression of c-MYC and PD-L1 in c-MYC-siRNA group was down-regulated significantly (all P<0.01). Conclusion: Over-expression of BIN1 could reduce the expression of PD-L1 by inactivating the c-MYC pathway, thereby inhibiting the immune escape of A549 cells.
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Project supported by the National Natural Science Foundation of China (No.81201607), the Outstanding Youth Foundation of Hebei Province (No. H2014206320), and the Foundation for High Level Talents of Hebei Province (No. A201401040)