MicroRNA‑642a‑5p inhibits colon cancer cell migration and invasion by targeting collagen type I α1

Abstract

The aim of the present study was to explore the mechanism by which microRNA (miR)‑642a‑5p regulates the migration and invasion of colon cancer cells via collagen type I α1 (COL1A1). The characteristics of miR‑642a‑5p and COL1A1 were analysed through bioinformatics. Cancer and normal tissues were collected from patients with colon cancer. miR‑642a‑5p‑ and COL1A1‑overexpressing cell lines were constructed by transfection. A dual‑luciferase reporter assay was used to verify the targeting of COL1A1 by miR‑642a‑5p. Cell Counting Kit‑8, wound healing and Transwell assays were used to detect cell viability, migration and invasion, respectively. Protein and mRNA expression levels were examined by western blotting and reverse transcription‑quantitative PCR, respectively. The results revealed that miR‑642a‑5p expression was significantly upregulated and COL1A1 expression was downregulated in patients with colon cancer. Low levels of miR‑642a‑5p and high levels of COL1A1 were associated with a poor prognosis in patients with colon cancer. miR‑642a‑5p directly targeted the 3'‑untranslated region of COL1A1 and inhibited COL1A1 expression. Overexpression of miR‑642a‑5p inhibited cell viability, migration, invasion and epithelial mesenchymal transition. Overexpression of COL1A1 promoted cell viability, migration, invasion and EMT, and partially reversed the inhibitory effects of miR‑642a‑5p on colon cancer cells. In conclusion, miR‑642a‑5p inhibited colon cancer cell migration, invasion and EMT by regulating COL1A1.

Keywords: colon cancer; microRNA-642a-5p; collagen type I α1; bioinformatics; migration; invasion; epithelial mesenchymal transition.

Figure 1.

miR-642a-5p expression is downregulated in colon cancer tissues. (A) Expression characteristics of miR-642a-5p in colon cancer from The Cancer Genome Atlas. (B) Association between miR-642a-5p expression and survival rate. miR, microRNA; COAD, colon adenocarcinoma; RPM, reads/counts of exon model per million mapped reads.

Figure 2.

COL1A1 expression is upregulated in patients with colon cancer and is associated with a poor prognosis by bioinformatics analysis in The Cancer Genome Atlas. (A) Intersection of the upregulated mRNAs as potential target genes of miR-642a-5p. (B) Genes that were significantly upregulated in colon cancer. (C) Expression characteristics of COL1A1 in colon cancer. (D) Association between COL1A1 expression and colon cancer stage. (E) Association between COL1A1 expression and survival rate. COL1A1, collagen type I α1; miR, microRNA; COAD, colon adenocarcinoma; FPKM, fragments per kilobase of exon model per million mapped fragments; TPM, transcripts per kilobase of exon model per million mapped reads.

Figure 3.

miR-642a-5p directly targets COL1A1. (A) Targeted binding site of miR-642a-5p and COL1A1. (B) miR-642a-5p expression in cells after transfection. (C) Results of the Dual-luciferase reporter assay. *P<0.05 vs. mimic-NC group. COL1A1, collagen type I α1; miR, microRNA; NC, negative control; WT, wild-type; Mut, mutant; Luc, luciferase.

Figure 4.

miR-642a-5p and COL1A1 expression is associated with prognosis in patients with colon cancer. Expression characteristics of (A) miR-642a-5p and (B) COL1A1 in patients with colon cancer. (C) Correlation between miR-642a-5p and COL1A1 expression in patients with colon cancer. Association between (D) miR-642a-5p or (E) COL1A1 expression and prognosis in patients with colon cancer. *P<0.05 vs. normal tissue group. COL1A1, collagen type I α1; miR, microRNA.

Figure 5.

miR-642a-5p inhibits colon cancer cell viability by targeting COL1A1. (A) COL1A1 protein expression in DLD-1 cells after COL1A1 overexpression. (B) mRNA expression levels of miR-642a-5p and COL1A1 in DLD-1 cells. (C) Protein expression levels of COL1A1 in DLD-1 cells. (D) DLD-1 cell viability in different groups. (E) COL1A1 protein expression in SW620 cells after COL1A1 overexpression. (F) mRNA expression levels of miR-642a-5p and COL1A1 in SW620 cells. (G) Protein expression levels of COL1A1 in SW620 cells. (H) SW620 cell viability in different groups. *P<0.05 vs. OE-NC + mimic-NC group; ^#P<0.05 vs. OE-NC + mimic-miR-642a-5p group. COL1A1, collagen type I α1; miR, microRNA; NC, negative control; OE, overexpression.

Figure 6.

miR-642a-5p inhibits colon cancer cell migration and invasion by targeting COL1A1. (A) DLD-1 cell migration (magnification, ×100) and (B) invasion (magnification, ×400) in each group. (C) SW620 cell migration (magnification, ×100) and (D) invasion (magnification, ×400) in each group. *P<0.05 vs. OE-NC + mimic-NC group; ^#P<0.05 vs. OE-NC + mimic-miR-642a-5p group. COL1A1, collagen type I α1; miR, microRNA; NC, negative control; OE, overexpression.

Figure 7.

miR-642a-5p inhibits epithelial mesenchymal transition via COL1A1. (A) mRNA and (B) protein expression levels of vimentin, N-cadherin and E-cadherin in DLD-1 cells. (C) mRNA and (D) protein expression levels of vimentin, N-cadherin and E-cadherin in SW620 cells. *P<0.05 vs. OE-NC + mimic-NC group; ^#P<0.05 vs. OE-NC + mimic-miR-642a-5p group. COL1A1, collagen type I α1; miR, microRNA; NC, negative control; OE, overexpression.