MiR-216a-5p/Hexokinase 2 axis regulates uveal melanoma growth through modulation of Warburg effect

https://doi.org/10.1016/j.bbrc.2018.05.069Get rights and content

Highlights

  • microRNA-216a-5p (miR-216a-5p) inhibits HK2 expression by directly targeting its 3′-UTR in uveal melanoma cells.

  • miR-216a-5p dampens glycolysis in uveal melanoma cells via inhibition of HK2.

  • miR-216a-5p expression is negatively correlated with HK2 expression and predicts better outcome in uveal melanoma patients.

Abstract

Hexokinase-2 (HK2), the initial as well as the rate-limiting step in glycolysis, is overexpressed in many human cancers, and correlates with poor clinical outcomes. Aerobic glycolysis is a hallmark of cancer, and drugs targeting its enzymes, including HK2, are being developed. However, the mechanisms of HK2 inhibition and the physiological significance of the HK2 inhibitors in cancer cells are rarely reported. Here, we show that microRNA-216a-5p (miR-216a-5p) inhibits HK2 expression by directly targeting its 3′-UTR in uveal melanoma cells. Through inhibition of HK2, miR-216a-5p dampens glycolysis by reducing HK activity, glucose uptake, lactate production, ATP generation, extracellular acidification rate (ECAR), and increasing oxygen consumption rate (OCR) in uveal melanoma cells. Importantly, glycolysis regulated by miR-216a-5p is critical for its regulating uveal melanoma tumor growth both in vitro and in vivo. miR-216a-5p expression is negatively correlated with HK2 expression and predicts better outcome in uveal melanoma patients. Our findings provide clues regarding the role of miR-216a-5p as a tumor suppressor in uveal melanoma through the inhibition of HK2. Targeting HK2 through miR-216a-5p could be a promising therapeutic strategy in uveal melanoma.

Section snippets

Aerobic glycolysis is involved in the regulation of uveal melanoma cell proliferation by miR-216a-5p

Our previous work demonstrated the tumor suppressive role of miR-216a-5p, and since aerobic glycolysis plays a critical role in cancer proliferation, we investigated if aerobic glycolysis plays a role in the inhibition of uveal melanoma cell proliferation mediated by miR-216a-5p. Anti-miR-216a-5p to promotes proliferation in A375 and MUM-2B cells, and more importantly, the glycolytic inhibitor 2-deoxy-d-glucose (2-DG) suppressed the ability of anti-miR-216a-5p to promote proliferation in A375

Discussion

The reprogramming of cell metabolism is an emerging hallmark of cancer. In aerobic glycolysis (also known as the Warburg effect), tumor cells depend on glycolysis for energy even under normoxic conditions, thus providing them with a growth advantage. Targeting the process of glycolysis has been demonstrated to be an effective way in controlling tumor growth and enhancing anticancer therapies. During the process of glycolysis, metabolic enzymes have attracted the most attentions for targets of

Cell lines, plasmids, lentivirus, RNA oligonucleotides, reagents

Human uveal melanoma cell lines and human embryonic kidney cell line HEK293T were purchased from the American Type Culture Collection (Manassas, VA, USA). Wild-type and mutated miR-216-5p putative targets on HK2 3′-UTR were cloned into pmiR-GLO dual-luciferase miRNA target expression vector (Promega). Lentiviral vectors expressing miR-216a or HK2 shRNA were constructed by cloning pre-miR-30a or HK2 short hairpin RNA fragments into pCDH and pSIH-H1-Puro (System Biosciences), respectively. The

Competing financial interest

The authors declare no competing financial interests.

Acknowledgements

This work was supported by National Natural Science Foundation (81770887, 61471039, 81471466, 81402345 and 81502264), Beijing Natural Science Foundation (7162180), Foundation in Education Ministry of Liaoning Province (JYTFUDF201749).

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