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. 2017 Mar;161(3):747-752.
doi: 10.1016/j.surg.2016.09.009. Epub 2016 Dec 2.

FX11 inhibits aerobic glycolysis and growth of neuroblastoma cells

Eric J Rellinger  1 , Brian T Craig  1 , Alexandra L Alvarez  1 , Haley L Dusek  1 , Kwang W Kim  1 , Jingbo Qiao  1 , Dai H Chung  2
Affiliations

FX11 inhibits aerobic glycolysis and growth of neuroblastoma cells

Eric J Rellinger et al. Surgery. 2017 Mar.

Abstract

Background: The MYC family of proteins promotes neuroblastoma tumorigenesis at least in part through the induction of aerobic glycolysis by promoting the transcription of key glycolytic enzymes, such as LDHA. FX11 is a selective inhibitor of LDHA that has demonstrated preclinical efficacy in adult cancers. Herein, we hypothesized that FX11 would inhibit aerobic glycolysis and block growth of neuroblastoma cells.

Methods: We surveyed 3 MYCN-single copy and 5 MYCN-amplified neuroblastoma cell lines to correlate C-MYC/N-MYC protein levels with LDHA expression. Cell viability was measured with FX11 using a tetrazolium-based assay. Cell cycle analysis using propidium iodide with flow cytometry was performed to evaluate for growth arrest. Immunoblotting demonstrated PARP and Caspase 3 cleavage as evidence of apoptosis.

Results: LDHA is frequently expressed in both MYCN--amplified and MYCN-single copy cell lines. N-MYC and C-MYC protein levels did not correlate with LDHA protein expression. FX11 inhibits aerobic glycolysis and growth in three MYCN-amplified and one MYCN-single copy neuroblastoma cell lines. FX11 induces modest G1 cell cycle arrest with selective induction of apoptosis.

Conclusion: Small molecule LDHA inhibition is capable of blocking aerobic glycolysis and growth of neuroblastoma cell lines in vitro and merits further in vivo evaluation of its preclinical efficacy in neuroblastomas.

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Conflict of interest statement

We have no financial or other interests, which may be construed as a conflict of interest.

Figures

Figure 1
Figure 1. FX11 inhibited aerobic glycolysis in neuroblastomas
A) Immunoblotting for C- MYC, N-MYC, and LDHA demonstrated a lack of correlation between basal MYC and LDHA protein levels in neuroblastoma cell lines β-actin was used for protein loading control. B) FX11 inhibited aerobic lactate production in neuroblastoma cell lines (*=p<0.05 for 10 μM FX11 treatment vs. control; ‡=p<0.05 for 20 μM FX11 treatment vs. control).
Figure 2
Figure 2. Neuroblastoma growth is blocked with FX11 treatment
Serial measurements of cell viability were performed using a tetrazolium-based assay (CCK-8) to evaluate the growth effects of daily FX11 treatment in MYCN-amplified (IMR-32, LAN-1, and BE(2)-C) and MYCN- single copy (SK-N-AS and SK-N-SH) cell lines (*=p<0.05 for 10 μM FX11 treatment vs. control; ‡=p<0.05 for 20 μM FX11 treatment vs. control).
Figure 3
Figure 3. FX11 induced modest G1 cell cycle arrest
Cell cycle analysis was completed using propidium iodide and flow cytometry 72 h following treatment of five experimental cell lines with daily FX11 (10 μM). Cell cycle analysis was completed in triplicate for each cell line with 10,000 events per replicate.
Figure 4
Figure 4. LDHA blockade with FX11 selectively induced apoptosis
Western blotting was performed to detect PARP and Caspase 3 cleavage products (1–4 d) following daily treatment with FX11 (10 μM). β-actin was used for protein loading control.
See this image and copyright information in PMC

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