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. 2020 Sep 8:12:8137-8147.
doi: 10.2147/CMAR.S253777. eCollection 2020.

MiR-138-5p Inhibits the Proliferation of Gastric Cancer Cells by Targeting DEK

Wei Zhang  1 , Kai Liao  1 , Dongning Liu  2
Affiliations

MiR-138-5p Inhibits the Proliferation of Gastric Cancer Cells by Targeting DEK

Wei Zhang et al. Cancer Manag Res. .

Retraction in

Abstract

Background: Increasing evidence suggests that microRNAs (miRNAs) play critical roles in cancer progression. Therefore, investigating the function of miRNAs that are aberrantly expressed in gastric cancer (GC) and characterizing the involved underlying mechanism are essential for the treatment of gastric cancer. MiR-138-5p was found to be down-regulated in multiple cancers, which acted as a tumor suppressor in cancer progression; however, whether and how miR-138-5p regulates the malignant behaviors of GC has not been fully understood.

Methods: The level of miR-138-5p in GC tissues and cell lines was detected by RT-qPCR. The effects of miR-138-5p on the growth of GC cells were evaluated by the in vitro Cell Counting Kit-8 (CCK-8) assay, cell apoptosis, cell cycle analysis, wound-healing assay, and in vivo xenograft mice model. The targets of miR-138-5p were predicted using the miRDB online tool, confirmed by luciferase report assay and Western blot.

Results: MiR-138-5p was frequently decreased in GC tissues and cell lines. Decreased expression of miR-138-5p was significantly associated with the lymph node metastasis of GC patients. Overexpression of miR-138-5p suppressed GC cell proliferation, migration, increased cell apoptosis as well as inhibited the tumor growth in vivo. DEK oncogene was predicted as a potential target of miR-138-5p. MiR-138-5p bound the 3'-UTR of DEK and inhibited the level of DEK in GC cells. Restoration of DEK abrogated miR-138-5p overexpression-mediated suppression of GC cell proliferation and cell cycle arrest.

Conclusion: Our results demonstrated the anti-cancer role of miR-138-5p in GC by targeting DEK, which suggested miR-138-5p as a potential therapeutic target for the treatment of patient with GC.

Keywords: DEK; gastric cancer; miR-138-5p.

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Conflict of interest statement

The authors declare that there are no conflicts of interest regarding this paper.

Figures

Figure 1
Figure 1
MiR-138-5p was down-regulated in GC. (A) RT-qPCR of miR-138-5p expression in GC tissues (n=50) and paired adjacent normal tissues (n=50). ***P<0.001 vs normal group. (B) The level of miR-138-5p was lower in patients with lymph node metastasis (n=20). (C) RT-qPCR analysis of miR-138-5p in normal cell GES-1 and GC cell lines MKN45, MKN28, NCI-N87 and AGS. ***P<0.001, **P<0.01 vs normal cells. Data were presented as the mean ± standard deviation.
Figure 2
Figure 2
Overexpression of miR-138-5p inhibited the proliferation, colony formation and induced apoptosis of GC cells. (A) The overexpression of miR-138-5p with the transfection of miR-138-5p mimics was detected by RT-qPCR. ***P<0.001 vs miR-NC group. (B and C) CCK-8 assay showed that overexpression of miR-138-5p significantly inhibited the proliferation of MKN45 and N87 cells compared with cells expressing miR-NC. ***P<0.001 vs miR-NC group. (D) The apoptosis of cells was detected by the staining of PI and Annexin V-FTIC. Overexpression of miR-138-5p induced a significant increase in the apoptosis (early and late apoptosis) of GC cells. ***P<0.001 vs miR-NC group. (E) Transfection of miR-138-5p inhibited the migration of GC cells in comparison with the control cells. ***P<0.001 vs miR-NC group. (F) Colony formation assay showed that miR-138-5p overexpression attenuated the proliferative capacity of both MKN45 and N87 cells. ***P<0.001 vs miR-NC group. (G) The expression of miR-138-5p in tumors infected with lentivirus expressed miR-138-5p mimics or miR-NC was confirmed by RT-qPCR analysis. ***P<0.001 vs miR-NC group. (H–J) Overexpressed miR-138-5p significantly inhibited the tumor volume (H and I) and weight (J) compared with that of control group harboring miR-NC, respectively. ***P<0.001 vs miR-NC group. Data were obtained from three independent experiments and presented as the mean ± standard deviation.
Figure 3
Figure 3
DEK was a target of miR-138-5p in GC. (A) Schematic of predicted binding sites of miR-138-5p in the 3ʹ-UTR of DEK predicted by the miRDB database. (B and C) Luciferase activity in GC cells co-transfected with luciferase vector expressing WT or MT 3ʹ-UTR of DEK and miR-138-5p mimics or miR-NC. ***P<0.001 vs miR-NC group. (D and E) The mRNA (D) and protein levels of DEK (E) in GC cells transfected with miR-138-5p mimics were decreased compared with the cells expressing miR-NC. ***P<0.001 vs miR-NC group. (F) The mRNA level of DEK in GC tissues (n=50) and paired adjacent normal tissues (n=50) was validated by RT-qPCR. The level of DEK was significantly reduced in GC tissues. ***P<0.001 vs adjacent normal tissues. (G) The expression of DEK was increased in GC patients with LNM (n=20) than those patients without LNM (n=30). ***P<0.001 vs LNM negative. (H) The IHC staining of DEK was increased in GC tissues especially those with lymph node metastasis. (I) The correlation between miR-138-5p and DEK in GC tissues was determined by the Spearman test. Data were obtained from three independent experiments and presented as the mean ± standard deviation.
Figure 4
Figure 4
Restoration of DEK reversed the suppressive function of miR-138-5p in GC. (A) The transfection efficiency of ectopic expressed Flag-DEK was confirmed by Western blot with anti-Flag antibody. (B and C) CCK-8 assay was performed to analyze the proliferation of MKN45 and N87 cells that transfected with miR-NC, miR-138-5p mimics or the combination with Flag-DEK. *P<0.05, ***P<0.001 vs cells transfected with miR-138-5p mimics alone. (D) Overexpression of DEK attenuated the G1 cell cycle arrest induced by miR-138-5p mimics in GC cells. ***P<0.001 vs cells transfected with miR-138-5p mimics alone. (E) Restoration of DEK significantly reversed miR-138-5p-induced apoptosis of MKN45 and N87 cells. ***P<0.001 vs cells transfected with miR-138-5p mimics alone. (F) Overexpression of miR-138-5p promoted the cleaved caspase-3/9 and the level of p21, while restoration of DEK attenuated the accumulation of cleaved caspase-3/9 and p21 in GC cells. Data were obtained from three independent experiments and presented as the mean ± standard deviation.
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