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Metformin Promotes Antitumor Immunity via Endoplasmic-Reticulum-Associated Degradation of PD-L1.

Mol Cell. 2018; 
Cha JH, Yang WH, Xia W, Wei Y, Chan LC, Lim SO, Li CW, Kim T, Chang SS, Lee HH, Hsu JL, Wang HL, Kuo CW, Chang WC, Hadad S, Purdie CA, McCoy AM, Cai S, Tu Y, Litton JK, Mittendorf EA, Moulder SL, Symmans WF, Thompson AM, Piwnica-Worms H, Chen CH, Khoo KH, Hung MC.
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Catalog Antibody 620 Molecular Cell 71, 606–620, August 16, 2018 STAR+METHODS KEY RESOURCES TABLE REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse CTLA-4 Bio X Cell Cat#BE0032; RRID:AB_1107798 hamster IgG Bio X Cell Cat#BE0091; RRID:AB_1107773 active caspase 3 Cell Signaling Technology Cat#9661L; RRID:AB_234118 CD8 BioRad Cat#MCA609G; RRID:AB_321407 GranzymeB R&D Systems Cat#AF1865; RRID:AB_2294988 PD-L1 Cell Signaling Technology Cat#13684; RRID:AB_2687655 PD-L1 LSBio Cat#338364; RRID:AB_597775 PD-L1 Abcam Cat#ab205921; RRID:AB_2687878 PD-L1 Cell Signaling Technology Cat#86744S PD-L1 GeneTex Cat#GTX104763; RRID:AB_1240586 mPD-L1 R&D Cat#AF1019; RRID:AB_354540 PD-L2 Cell Signaling Technology Cat#82723 B7H3 Cell Signaling Technology Cat#14058 PVR Cell Signaling Technology Cat#13544 VISTA Cell Signaling Technology Cat#54979 Galectin-9 Life Technology Cat#50-9116-41; RRID:AB_2574323 CD86 Abcam Cat#Ab53004; RRID:AB_869050 HSP90B1 Novus Biologicals Cat#NBP2-42379 GM130 BD Biosciences Cat#610823; RRID:AB_398142 Alexa Fluor 488 Giantin BioLegend Cat#908701; RRID:AB_256089 IRE1a1 Cell Signaling Technology Cat#3294 TNG46 Novus Biologicals Cat#NB110-62093; RRID:AB_965643 AMPKa1 R&D Systems Cat#AF3197; RRID:AB_2169570 AMPKa2 R&D Systems Cat#AF2850; RRID:AB_2169572 AMPKa Cell Signaling Technology Cat#5831S; RRID:AB_10622186 AMPKa Thr172-p Cell Signaling Technology Cat#2535S; RRID:AB_331250 Flag-tag Cell Signaling Technology Cat#14793S SEL1L Sigma-Aldrich Cat#S3699; RRID:AB_1856660 HDR1 Cell Signaling Technology Cat#14773S ERELC1 Abcam Cat#ab181166 OS9 Cell Signaling Technology Cat#12497S HIS-tag GenScript Cat#A00186; RRID:AB_914704 GST-tag Bethyl Lab Cat#A190-122A; RRID:AB_67419 HA-tag Cell Signaling Technology Cat#3724S; RRID:AB_1549585 GAPDH Cell Signaling Technology Cat#2118; RRID:AB_561053 a-Tubulin Sigma-Aldrich Cat#T5168; RRID:AB_477579 b-Actin Sigma-Aldrich Cat#A2228; RRID:AB_476697 phospho-huPD-L1 Ser195 This paper N/A CD3 eBioscience Cat#16-0037; RRID:AB_468854 CD16/CD32 BioLegend Cat#40477 PD-L1-APC BD Cat#564715 CD3-PreCP BioLegend Cat#100325 CD8-APC/Cy7 BioLegend Cat#100713 (Continued on next page) Molecular Cell 71, 606–620.... Immunoblotting was per- formed with primary antibodies against PD-L1 (13684, 1:1,500; Cell Signaling Technology), PD-L1 (338364, 1:1,000; LSBio), AMPKa (5831S, 1:2,000; Cell Signaling Technology), AMPKa1 (AF3197, 1:2000; R&D Systems), AMPKa2 (AF2850, 1:2000; R&D Systems), AMPKa Thr172-p (2535S, 1:1,000; Cell Signaling Technology), Flag-tag (14793S, 1:2,000; Cell Signaling Technology), SEL1L (S3699, 1:5,000; Sigma-Aldrich), HDR1 (14773S, 1:2000; Cell Signaling Technology), ERELC1 (ab181166, 1:3,000; Abcam, San Fran- cisco, CA, USA), OS9 (12497S, 1:1,000; Cell Signaling Technology), HIS-tag (A00186, 1:5000; GenScript), GST-tag (A190-122A, 1:10000; Bethyl Lab), HA-tag (3724S, 1:3,000; Cell Signaling Technology), GAPDH (2118, 1:3,000; Cell Signaling Technology), a-Tubulin (B-5-1-2, 1:5,000; Sigma-Aldrich), b-Actin (A2228, 1:10,000; Sigma-Aldrich). Get A Quote

摘要

Metformin has been reported to possess antitumor activity and maintain high cytotoxic T lymphocyte (CTL) immune surveillance. However, the functions and detailed mechanisms of metformin's role in cancer immunity are not fully understood. Here, we show that metformin increases CTL activity by reducing the stability and membrane localization of programmed death ligand-1 (PD-L1). Furthermore, we discover that AMP-activated protein kinase (AMPK) activated by metformin directly phosphorylates S195 of PD-L1. S195 phosphorylation induces abnormal PD-L1 glycosylation, resulting in its ER accumulation and ER-associated protein degradation (ERAD). Consistently, tumor tissues from metformin-treated breast cancer patients ... More

關鍵詞

ER accumulation; ERAD; PD-L1; cancer immunotherapy; glycosylation; immune checkpoint blockade; metformin
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