Research Paper Volume 12, Issue 12 pp 11685—11697
Long noncoding RNA KCNQ1OT1 promotes colorectal carcinogenesis by enhancing aerobic glycolysis via hexokinase-2
- 1 Department of General Surgery, Qilu Hospital of Shandong University, Jinan 250012, China
Received: March 4, 2020 Accepted: April 20, 2020 Published: June 21, 2020https://doi.org/10.18632/aging.103334
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Copyright © 2020 Chen et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
In this study, we investigated the mechanistic role and prognostic significance of the long coding RNA (lncRNA) KCNQ1OT1 in colorectal cancer (CRC). KCNQ1OT1 levels were significantly higher in CRC tissues than adjacent normal colorectal tissues (n=79). High KCNQ1OT1 expression correlated with poorer prognosis in CRC patients. KCNQ1OT1-silenced CRC cells showed reduced proliferation, colony formation, extracellular acidification, and lactate and glucose secretion. This suggests KCNQ1OT1 promotes CRC cell proliferation by increasing aerobic glycolysis. RNA pull-down assays with biotinylated KCNQ1OT1 followed by mass spectrometry analysis showed that KCNQ1OT1 directly binds to hexokinase 2 (HK2). This was confirmed by RNA immunoprecipitation assays using anti-hexokinase 2 antibody. HK2 protein levels were reduced in KCNQ1OT1 knockdown CRC cells, but were restored by treatment with the proteasomal inhibitor MG132. KCNQ1OT1 knockdown CRC cells also showed higher ubiquitinated-HK2 levels, suggesting KCNQ1OT1 enhances aerobic glycolysis by stabilizing HK2. HK2 overexpression in KCNQ1OT1 knockdown CRC cells restored proliferation and aerobic glycolysis. KCNQ1OT1 levels correlated positively with HK2 expression and prognosis in CRC patients. These findings show that KCNQ1OT1 promotes colorectal carcinogenesis by increasing aerobic glycolysis through HK2.
DFS: Disease-free survival; IHC: Immunohistochemical staining; ncRNA: Noncoding RNA; OS: Overall survival; qRT-PCR: Quantitative real-time PCR; RIP: RNA immunoprecipitation; ROC: Receiver operating characteristic; Ctrl: Control; TNM: Tumor-node-metastasis; MS: mass spectrometry; CCK8: Cell Counting Kit-8; DMEM: Dulbecco’s Modified Eagle Medium; HK2: Hexokinase II; Sh-RNA: short hairpin RNA; CHX: Cycloheximide; ECAR: Extracellular acidification rate.