USP11 stabilizes RALY to promote FXYD5-mediated aerobic glycolysis and aggravate pancreatic cancer progression.

BACKGROUND

Pancreatic cancer (PC) is a highly malignant and aggressive gastrointestinal malignancy with a poor prognosis for patients. Aerobic glycolysis serves as a critical metabolic driver of PC progression. Our study aims to elucidate the mechanism by which FXYD5 regulates aerobic glycolysis in PC.

METHODS

Cell viability was assessed using CCK-8. Cell proliferation was tested by colony formation assay. Lactic acid production and ATP levels were measured by commercial kits. Extracellular acidification rate (ECAR) was detected using the Seahorse XF96 analyzer. RNA immunoprecipitation (RIP) was employed to validate the binding of RALY to FXYD5 mRNA. The interaction between RALY and USP11, as well as the ubiquitinated level of RALY, were probed by co-immunoprecipitation (Co-IP) assay. A subcutaneous xenograft tumor model was established to verify the in vitro findings.

RESULTS

Knockdown of FXYD5 suppressed lactic acid production, ECAR, ATP levels, and the expression of aerobic glycolysis-related key markers (GLUT1 and HK2) in PC cells. RALY directly bound to FXYD5 mRNA to promote its stability in PC cells. RALY facilitated aerobic glycolysis to increase PC tumor growth by up-regulating FXYD5 expression in vitro and in vivo. Deubiquitinating enzyme USP11 reduced the ubiquitinated level of RALY to maintain its protein stability. USP11 knockdown restrained aerobic glycolysis of PC cells, which was reversed by RALY overexpression.

CONCLUSION

USP11 stabilized FXYD5 mRNA by promoting RALY deubiquitinated modification to increase PC cell aerobic glycolysis and exacerbate tumor progression.