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. 2019 Jan;17(1):555-563.
doi: 10.3892/ol.2018.9595. Epub 2018 Oct 18.

NANOGP8 expression regulates gastric cancer cell progression by transactivating DBC1 in gastric cancer MKN-45 cells

Li Li  1   2 , Ru Feng  3 , Sujuan Fei  2 , Jiang Cao  4 , Qinqin Zhu  2 , Guozhong Ji  5 , Jianwei Zhou  1
Affiliations

NANOGP8 expression regulates gastric cancer cell progression by transactivating DBC1 in gastric cancer MKN-45 cells

Li Li et al. Oncol Lett. 2019 Jan.

Abstract

NANOGP8 is one of the NANOG pseudogenes and is expressed together with NANOG in multiple tumor tissues and cell lines. The biological functions of NANOGP8 in progression of gastric cancer are unclear. In the present study, the role of NANOGP8 was investigated in gastric cancer cells. The gathered data demonstrated that NANOG expression in both mRNA and protein was elevated in gastric cancer cell lines relative to a normal gastric epithelial cell line. Downregulation of NANOGP8 inhibited cell proliferation and increased apoptosis in human gastric carcinoma cell lines. Furthermore, silencing of NANOGP8 suppressed tumor growth in vivo. Interestingly, it was identified that deleted in breast cancer 1 (DBC1) expression was also markedly downregulated following NANOGP8 knockdown. DNA microarray and dual-luciferase assays further indicated that NANOGP8 may bind to the DBC1 promoter region and regulate DBC1 expression. Therefore, the gathered data provided evidence that NANOGP8 contributes to progression of gastric cancer via DBC1 and may have potential translational significance.

Keywords: apoptosis; gastric cancer; proliferation.

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Figures

Figure 1.
Figure 1.
NANOG mRNA and protein expression in gastric cancer cell lines and surgical specimens. (A) NANOG mRNA expression levels in gastric cancer cell lines. (B) NANOG protein expression levels in gastric cancer cell lines. (C) NANOG mRNA expression levels in surgical specimens. M, marker, NANOG, nanoghomeobox.
Figure 2.
Figure 2.
Downregulation of NANOGP8 expression in human gastric cancer cells. (A) The effect of NANOGP8 silencing on MKN-45 cell proliferation. (B) The effect of NANOGP8 silencing on MKN-45 cell apoptosis. (C) The effect of NANOGP8 silencing on MKN-45 proliferation (magnification, ×400). *P<0.05 vs. sh-control. NANOG, nanoghomeobox; OD, optical density.
Figure 3.
Figure 3.
Tumors from nude mice injected with sh-control cells or shNANOG-2 cells.
Figure 4.
Figure 4.
The effect of NAONGP8 silencing on tumor growth in nude mice. (A) Tumor weight. (B) Tumor volume. *P<0.05 vs. sh-control.
Figure 5.
Figure 5.
DBC1 mRNA and protein expression in gastric cancer cell lines and surgical specimens. (A) DBC1 mRNA expression levels in gastric cancer cell lines. (B) DBC1 protein expression levels in surgical specimens. DBC1, deleted in breast cancer 1.
Figure 6.
Figure 6.
mRNA expression of NANOGP8 and DBC1 in sh-control, sh-NANOGP8, sh-DBC1, and shDBC1+NANOGP8 cells. *P<0.05 vs. sh-control. NANOG, nanoghomeobox; DBC1, deleted in breast cancer 1.
Figure 7.
Figure 7.
Downregulation of NANOGP8 or DBC1 expression in human gastric cancer cells. (A) The effect of NANOGP8 or DBC1 silencing on MKN-45 cell proliferation. (B) The effect of NANOGP8 or DBC1 silencing on MKN-45 cell apoptosis. (C) The effect of NANOGP8 or DBC1 silencing on MKN-45 cells proliferation. *P<0.05 vs. sh-control. OD, optical density; NANOG, nanoghomeobox; DBC1, deleted from breast cancer 1.
Figure 8.
Figure 8.
The NANOGP8 binding site within the DBC1promoter region. (A) Motif sequence signature of NANOGP8. (B) The predicted binding site of the NANOGP8 transcription factor. NANOG, nanoghomeobox.
Figure 9.
Figure 9.
Luciferase assay detection of DBC1 promoter activation by NANOGP8. *P<0.05 vs. pGL3-Basic; SP<0.05 vs. pcDNA3.1(+) co-transfected with pGL3-DBC1. DBC1, deleted from breast cancer 1.
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