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. 2021 Jan 22;21(1):70.
doi: 10.1186/s12935-020-01743-5.

Hydrogen inhibits the proliferation and migration of gastric cancer cells by modulating lncRNA MALAT1/miR-124-3p/EZH2 axis

Baocheng Zhu #  1 , Hengguan Cui #  1 , Weiqiang Xu  2   3
Affiliations

Hydrogen inhibits the proliferation and migration of gastric cancer cells by modulating lncRNA MALAT1/miR-124-3p/EZH2 axis

Baocheng Zhu et al. Cancer Cell Int. .

Abstract

Background: Gastric cancer is one of the most prevalent and deadly malignancies without efficient treatment option. This study aimed to investigate the effect of hydrogen gas on the behavior of gastric cancer cells.

Methods: Gastric cancer cell lines MGC-803 and BGC-823 were treated with or without H2 /O2 gas mixture (66.7%:33.3% v/v). Proliferation and migration were assessed by MTT and scratch wound healing assays respectively. The expression of lncRNA MALAT1, miR-124-3p, and EZH2 was analyzed by real-time quantitative PCR and/or western blot. Tumor growth was estimated using xenograft mouse model.

Results: H2 gas significantly inhibited gastric tumor growth in vivo and the proliferation, migration, and lncRNA MALAT1 and EZH2 expression of gastric cancer cells while upregulated miR-124-3p expression. LncRNA MALAT1 overexpression abolished all the aforementioned effects of H2. LncRNA MALAT1 and miR-124-3p reciprocally inhibited the expression of each other. MiR-124-3p mimics abrogated lncRNA MALAT1 promoted EZH2 expression and gastric cancer cell proliferation and migration.

Conclusions: These data demonstrated that H2 might be developed as a therapeutics of gastric cancer and lncRNA MALAT1/miR-124-3p/EZH2 axis could be a target for intervention.

Keywords: EZH2; Gastric cancer; Hydrogen; LncRNA MALAT1; Migration; Proliferation; miR-124-3p.

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Conflict of interest statement

The authors declare no potential conflict of interest.

Figures

Fig. 1
Fig. 1
Gastric cancer cell proliferation and migration were inhibited by H2 gas. a MGC-803 and BGC-823 cells were cultured with or without H2 for 24 h and assessed by MTT assay. b MGC-803 and BGC-823 cell migration was assayed by scratch wound healing assay. c Quantitative analysis of migration data. Scale bar: 25 µm. Ctrl, Normal culture conditions. *p < 0.01 compared to Ctrl
Fig. 2
Fig. 2
The expression of lncRNA MALAT1 and its target genes in gastric cancer cells was inhibited by H2. a Heatmap of genes differently expressed in MGC-803 cells cultured with or without H2. b Transcript levels of lncRNA MALAT1, EZH2, were analyzed by quantitative real-time PCR. c Protein levels of EZH2 were assessed by immunoblot. CK & Ctrl, Normal culture conditions. *p < 0.01 compared to Ctrl
Fig. 3
Fig. 3
H2 inhibited gastric cancer cell proliferation and migration through regulating lncRNA MALAT1. a Transcript levels of lncMALAT1 and EZH2 of MGC-803 cells were measured by quantitative real-time PCR. b Protein level of EZH2 was assessed by immunoblot. c MGC-803 and BGC-823 cells transfected with or without lncRNA MALAT1 expression vector were cultured with or without H2 for 24 h and assessed by MTT assay. d Scratch wound healing assay was used to examine the effects of H2 and lncRNA MALAT1 on MGC-803 and BGC-823 cell migration. Scale bar: 25 µm. Ctrl, Normal culture conditions. *p < 0.01 compared to Ctrl; #p < 0.01 compared to MALAT1; &p < 0.01 compared to H2
Fig. 4
Fig. 4
LncRNA MALAT1 mediated the effects of H2 on miR-124-3p expression in gastric cancer cells. a The binding sites of miR-124-3p within lncRNA MALAT1 transcript sequence predicted using LncBase Predicted v.2 of Diana Tools. b The expression level of miR-124-3p in MGC-803 and BGC-823 cells transfected with or without lncRNA MALAT1 expression vector and cultured with or without H2 for 24 h was assayed by quantitative real-time PCR. *p < 0.01 compared to control; #p < 0.01 compared to MALAT1; &p < 0.01 compared to H2
Fig. 5
Fig. 5
The upregulation of EZH2 by lncRNA MALAT1 was antagonized by miR-124-3p. a The miR-124-3p binding site in EZH2 3’UTR predicted using TargetScan. b The expression levels of lncRNA MALAT1, miR-124-3p, and EZH2 after lncRNA MALAT1 and/or miR-124-3p overexpression were analyzed by quantitative real-time PCR. c The effects of lncRNA MALAT1 and miR-124-3p on EZH2 protein level were examined by western blot. Ctrl, Normal culture conditions. *p < 0.01 compared to control; #p < 0.01 compared to MALAT1; &p < 0.01 compared to 124-3p
Fig. 6
Fig. 6
miR-124-3p blocked lncRNA MALAT1 induced proliferation and migration of gastric cancer cells. a MGC-803 cells transfected with lncRNA MALAT1 and/or miR-124-3p were cultured for 24 h and assessed by MTT assay. b The migration of MGC-803 and BGC-823 cells with lncRNA MALAT1 and/or miR-124-3p overexpression was assayed by scratch wound healing assay. Scale bar: 25 µm. Ctrl, Normal culture conditions. *p < 0.01 compared to control; #p < 0.01 compared to MALAT1; &p < 0.01 compared to 124-3p
Fig. 7
Fig. 7
H2 inhibited gastric tumor growth via lncRNA MALAT1/miR-124-3p axis. BGC-823 cells stably expressing lncRNA MALAT1 and/or miR-124-3p were grafted into nude mice subcutaneously. Mice were then exposed to H2 for 2 h daily for 5 weeks. a Picture of representative tumors (upper panel) was shown and tumor size (lower panel) was measured. b EZH2 protein level of tumor tissue was evaluated by western blot. *p < 0.01 compared to Ctrl; ^p < 0.01 compared to H2 + 124-3p; #p < 0.01 compared to H2 + MALAT1; &p < 0.01 compared to H2
Fig. 8
Fig. 8
A proposed model for how H2 inhibits gastric cancer growth and invasion. Please refer to the text for details. formula image activation; formula image inhibition
See this image and copyright information in PMC

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