Lactylation stabilizes DCBLD1 activating the pentose phosphate pathway to promote cervical cancer progression

Abstract

Background Discoidin, CUB, and LCCL domain-containing type I (DCBLD1) is identified as an oncogene involved in multiple regulation of tumor progression, but specific mechanisms remain unclear in cervical cancer. Lactate-mediated lactylation modulates protein function. Whether DCBLD1 can be modified by lactylation and the function of DCBLD1 lactylation are unknown. Therefore, this study aims to investigate the lactylation of DCBLD1 and identify its specific lactylation sites. Herein, we elucidated the mechanism by which lactylation modification stabilizes the DCBLD1 protein. Furthermore, we investigated DCBLD1 overexpression activating pentose phosphate pathway (PPP) to promote the progression of cervical cancer. Methods DCBLD1 expression was examined in human cervical cancer cells and adjacent non-tumorous tissues using quantitative reverse transcription-polymerase chain reaction, western blotting, and immunohistochemistry. In vitro and in vivo studies were conducted to investigate the impact of DCBLD1 on the progression of cervical cancer. Untargeted liquid chromatography-tandem mass spectrometry (LC–MS/MS) metabolomics studies were used to characterize DCBLD1-induced metabolite alterations. Western blot, immunofuorescence and transmission electron microscopy were performed to detect DCBLD1 degradation of G6PD by activating autophagy. Chromatin immunoprecipitation, dual luciferase reporter assay for detecting the mechanism by which lactate increases DCBLD1 transcription. LC–MS/MS was employed to verify specific modification sites within the DCBLD1 protein. Results We found that lactate increased DCBLD1 expression, activating the PPP to facilitate the proliferation and metastasis of cervical cancer cells. DCBLD1 primarily stimulated PPP by upregulating glucose-6-phosphate dehydrogenase (G6PD) expression and enzyme activity. The mechanism involved the increased enrichment of HIF-1α in the DCBLD1 promoter region, enhancing the DCBLD1 mRNA expression. Additionally, lactate-induced DCBLD1 lactylation stabilized DCBLD1 expression. We identified DCBLD1 as a lactylation substrate, with a predominant lactylation site at K172. DCBLD1 overexpression inhibited G6PD autophagic degradation, activating PPP to promote cervical cancer progression. In vivo, 6-An mediated inhibition of G6PD enzyme activity, inhibiting tumor proliferation. Conclusions Our findings revealed a novel post-translational modification type of DCBDL1, emphasizing the significance of lactylation-driven DCBDL1-mediated PPP in promoting the progression of cervical cancer. Graphical Abstract Schematic illustration of DCBLD1-induced G6PD-mediated reprogramming of PPP metabolism in promoting cervical cancer progression.

Lactylation: the novel histone modification influence on gene expression, protein function, and disease

“…The K673 site of MRE11 protein is lactated by CBP to promote homologous recombination repair, which plays a key role in tumor chemotherapy resistance [ 72 ]. In cervical cancer, DCBLD1 with a lactylation site at K172 which can stabilize its expression, in order to inhibit G6PD autophagic degradation, activating pentose phosphate pathway (PPP) to promote the progression [ 73 ].…”

Lactylation Modification in Cardiometabolic Disorders: Function and Mechanism

“…Chen showed that MRE11, an important protein involved in homologous recombination (HR), is modified by lactylation at K673 by the CBP acetyltransferase in reaction to DNA damage, and this process relies on ATM phosphorylation [129]. Meng employed LC-MS/MS to confirm certain modification sites in the DCBLD1 protein, revealing that DCBLD1 is a substrate for lactylation, with the major lactylation site located at K172 [130].…”

Lactylation: The emerging frontier in post-translational modification

“…Inactive von Hippel-Lindau (VHL) promotes the progression of clear cell renal cell carcinoma by initiating a positive feedback loop between histone lactylation and platelet-derived growth factor receptor β (PDGFRβ) signaling ( Yang J. et al, 2022 ). Recent studies have demonstrated that lactylation modification plays a significant role in promoting tumor progression in various types of cancer, including prostate cancer ( Luo et al, 2022 ), colorectal cancer ( Wang et al, 2022a ; Li X. M. et al, 2024 ), ocular melanoma ( Gu et al, 2024 ), gastric cancer ( Sun et al, 2023 ), bladder cancer ( Xie et al, 2023 ) and cervical cancer ( Meng et al, 2024 ). By binding to lactate, alanyl-tRNA synthetase 1 (AARS1) is able to catalyze the formation of lactate-AMP complex, thereby promoting p53 lactylation and tumor growth ( Zong et al, 2024 ).…”