Aberrant STAT3 signaling in pancreatic cancer requires the lncRNA NEAT1 regulated by the metalloprotease-disintegrin ADAM8 via miR-181a-5p
Author:
Gao Yutong1, Zandieh Kimia1, Zhao Kai1, Khizanishvili Natalia1, DiFazio Pietro1, Yu Xiangdi2, Schulte Leon1, Aillaud Michelle1, Chung Ho-Ryun1, Ball Zachary3, Meixner Marion1, Bauer Uta-Maria1, Bartsch Detlef Klaus1, Buchholz Malte1, Lauth Matthias1, Nimsky Christopher1, Cook Lena1, Bartsch Jörg W.1
Affiliation:
1. Philipps University of Marburg 2. Guizhou University 3. Rice University
Abstract
Abstract
Purpose
Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal cancers. Several studies demonstrate that ADAM8 and STAT3 are critical for PDAC progression and potential therapeutic targets.
Methods
TGCA, microarray, and immunohistochemistry data from a PDAC cohort were used for clinical analyses. Panc89 cells with ADAM8 knockout, re-expression of ADAM8 mutants, and Panc1 cells overexpressing ADAM8 were generated. Gene expression analyses of ADAM8, STAT3, long non-coding (lnc) RNA NEAT1, miR-181a-5p and ICAM1 were performed by quantitative PCR. Subcellular fractionation quantified NEAT1 expression in cytoplasm and nucleus of PDAC cell lines. Cell proliferation, scratch, and invasion assays were performed to detect growth rate, migration and invasion capabilities of cells. Gain and loss of function experiments were carried out to investigate the biological effects of lncRNA NEAT1 and miR-181a-5p on PDAC cells and downstream genes. Dualluciferase reporter gene assay determined interaction and binding sites of miR-181a-5p in lncRNA NEAT1. Pull down assays, RNA binding protein immunoprecipitation (RIP), and ubiquitination assays explored the molecular interaction between lncRNA NEAT1 and STAT3.
Results
High ADAM8 expression causes aberrant STAT3 signaling and activation of downstream genes in PDAC cells and is positively correlated with NEAT1 expression. NEAT1 binding to STAT3 was confirmed and prevents STAT3 degradation in the proteasome as increased degradation of STAT3 was observed in ADAM8 knockout cells and cells treated with bortezomib. Furthermore, miRNA-181a-5p whose expression is controlled by ADAM8, regulates NEAT1 expression by direct binding to the NEAT1 promoter.
Conclusion
ADAM8 regulates intracellular STAT3 levels via miR-181a-5p and NEAT1 in pancreatic cancer.
Publisher
Springer Science and Business Media LLC
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