LncRNA HOXC-AS1 Sponges miR-99a-3p and Upregulates MMP8, Ultimately Promoting Gastric Cancer

SIMPLE SUMMARY: Long noncoding RNAs, including the HOXC Cluster Antisense RNA 1 (HOXC-AS1), are reported to be critical during the occurrence and progression of gastric cancer.

We examined cells and tissues for the expression of HOXC-AS1 and correlated the expression levels with the disease specific survival of the gastric cancer patients.

We also identified the interaction between HOXC-AS1 and miR-99a-3p, as well as matrix metalloproteinase 8 (MMP8) by dual-luciferase reporter gene assays.

Western blot and qRT-PCR were conducted to verify the alteration in expression levels, while Cell Counting Kit-8 assay and colony formation assay were performed to explore the influences on gastric cancer cells.

Overexpression of HOXC-AS1 would accordingly sponge greater quantities of miR-99a-3p, leading to the upregulation of MMP8, eventually facilitating the progress of gastric cancer.

ABSTRACT: Gastric cancer (GC) is among the most lethal tumors worldwide.

Long noncoding RNAs (lncRNAs) are reported to be critical during the occurrence and progression of malignancies.

The HOXC cluster antisense RNA 1 (HOXC-AS1) has been suggested to participate in the genesis and development of GC.

Therefore, we examined GC cells and tissues for the expression of HOXC-AS1 and correlated the expression levels with the disease specific survival of the patients, finding that HOXC-AS1 was overexpressed and probably had a tendency of leading to a poor prognosis.

The Cell Counting Kit-8 assay and colony formation assay were then performed under knockdown of HOXC-AS1, revealing that cell proliferation of GC was distinctly decreased.

Afterwards, miR-99a-3p was predicted to bind with HOXC-AS1 by DIANA tools.

We carried out dual-luciferase reporter gene assays to identify the interaction between them.

After knockdown of HOXC-AS1, miR-99a-3p was clearly overexpressed in GC cells.

In addition, matrix metalloproteinase 8 (MMP8) was shown to be combined with miR-99a-3p using TargetScan.

Similar experiments, along with western blot, were conducted to validate the correlation between miR-99a-3p and MMP8.

Finally, rescue experiments for CCK-8 were completed, disclosing that HOXC-AS1 promoted cell progression of GC through sponging miR-99a-3p followed by subsequent upregulation of MMP8.