A novel lnc-LAMC2-1:1 SNP promotes colon adenocarcinoma progression by targeting miR-216a-3p/HMGB3

Abstract

Single nucleotide polymorphisms (SNPs) was associated with altering the secondary structure of long non-coding RNA (lncRNA). Increasing reports showed that lnc-LAMC2-1:1 SNP played an important role in cancer development and invasion. This study is to elucidate the molecular function of lnc-LAMC2-1:1 SNP rs2147578 promoting tumor progression in colon adenocarcinoma (COAD). In this study, we found that the lnc-LAMC2-1:1 SNP rs2147578 was upregulated in COAD cell lines. Furthermore, lnc-LAMC2-1:1 SNP rs2147578 promoted colon cancer migration, invasion, and proliferation. Interestingly, lnc-LAMC2-1:1 SNP rs2147578 positively regulated HMGB3 expression via miR-216a-3p in colon cancer cells. Functional enrichment analysis showed that targeting genes of miR-216a-3p were enriched in regulating the pluripotency of stem cells, MAPK signaling pathway, TNF signaling pathway, neurotrophin signaling pathway, relaxin signaling pathway, and FoxO signaling pathway. Tumor Immune Estimation Resource (TIMER) database revealed that there was a significantly positive correlation between HMGB3 expression and the infiltration of CD8⁺ T cells, B cells, neutrophils, macrophages, and CD4⁺ T cells. Finally, HMGB3 overexpression was validated in external data. In conclusions, lnc-LAMC2-1:1 SNP rs2147578 was involved in promoting COAD progression by targeting miR-216a-3p/HMGB3, and this study will provide a novel molecular target for COAD.

Keywords: Colon adenocarcinoma; HMGB3; Progression; lnc-LAMC2-1:1 SNP; miR-216a-3p.

Figure 1

The lnc-LAMC2-1:1 SNP is increased and promotes colon cancer progression. Knockdown of lnc-LAMC2-1:1 SNP inhibits (a) proliferation, (b) migration, and (C) invasion in COAD. ∗P < 0.05, ∗∗P < 0.01 compared with the indicated control group.

Figure 2

The lnc-LAMC2-1:1 SNP is a sponge of miR-216a-3p. Bar graphs of (a) and (b) show the relative luciferase activity of vectors containing SW480. A dual-luciferase reporter assay was performed, and the co-transfection of lnc-LAMC2-1:1 SNP and miR-216a-3p reduced the luciferase activity.

Figure 3

The regulatory network of miR-126-3p and PPI (a) The network of miR-126-3p target genes. Red indicates miR-126-3p, and green indicates genes. (b) protein-protein interaction of target genes.

Figure 4

GO and KEGG functional enrichment analysis (a) GO enrichment significance items (b) Gene ratio and KEGG pathway items. Red indicates upregulated genes, and green indicates downregulated genes.

Figure 5

The lnc-LAMC2-1:1 SNP positively regulates HMGB3 by sponging miR-126-3p (a) mRNA expression of HMGB3 in SW480 cells. Cells were transfected with mimics of NC, miR-126-3p mimics, ASO-NC, and ASO-miR-126-3p. (b) mRNA expression of HMGB3 in SW480 cells. Cells were transfected with pcDNA3.1, lnc-LAMC2-1:1 SNP, and lnc-LAMC2-1:1-wt. (c) Complementary sequences of HMGB3 and miR-126-3p in the StarBase database. (d) A dual-luciferase reporter assay was performed, and the co-transfection of miR-126-3p and HMGB3 reduced the luciferase activity.

Figure 6

Correlation of HMGB3 expression with immune cell infiltration. Association of the expression of (a) CD8⁺ T cells, (b) CD4⁺ T cells, (c) macrophages, (d) neutrophils, (e) dendritic cells and (f) B cells with HMGB3 expression were shown in this study.

Figure 7

External data validation of HMGB3 expression. HMGB3 expression was validated in (a) GSE8671, (b) GSE9348, and (c) GEPIA database and (d) TCGA database. (e) and (f) show the expression of HMGB3 in COAD tissues on immunohistochemistry staining.