Silencing of long non-coding RNA HCP5 inhibits proliferation, invasion, migration, and promotes apoptosis via regulation of miR-299-3p/SMAD5 axis in gastric cancer cells

Abstract

Gastric cancer (GC) is a common malignant gastrointestinal tumor with high mortality. Previous study has reported that the overexpression of lncRNA HCP5 was observed in gastric cancer tissues. The purpose of this study was to investigate the molecular mechanism underlying the effect of lncRNA HCP5 on the proliferative, migratory, and invasive abilities of GC cells. The relative mRNA expression of HCP5, miR-299-3p, and SMAD5 were determined by RT-qPCR. The expressions of proteins associated with apoptosis and invasion were detected by western blot. The interaction of HCP5 with miR-299-3p and SMAD5 with miR-299-3p was confirmed by luciferase reporter assay. The cellular behaviors of AGS cells were, respectively, detected by CCK-8 assays, colony formation assays, migration and invasion assays, and flow cytometry. In our study, lncRNA HCP5 was highly expressed in GC cell lines compared with normal gastric epithelial cell. LncRNA HCP5 silencing inhibited AGS cells proliferation, migration, and invasion, while promoted cell apoptosis. Moreover, miR-299-3p downregulation could abolish the effect of HCP5 knockdown on cellular behaviors of AGS cells. Interestingly, SMAD5 is identified as the downstream target of miR-299-3p, and its expression was inhibited by miR-299-3p. More importantly, SMAD5 silencing inhibited proliferation, migration, and invasion of GC cells, and promoted cell apoptosis. In a word, lncRNA HCP5 silencing inhibits GC cell proliferation, invasion, and migration while promoting its apoptosis via regulation of miR-299-3p/SMAD5 axis. Hence, lncRNA HCP5 could be a novel and promising target for GC treatment.

LncRNA HCP5 as a potential therapeutic target and prognostic biomarker for various cancers: a meta‑analysis and bioinformatics analysis

“… In esophageal squamous cell carcinoma, HCP5 promoted cellular activities via modulating the miR-139-5p/PDE4A pathway and stimulating the PI3K/AKT/mTOR signaling pathway [ 35 ]. In gastric cancer, HCP5 was highly expressed in gastric cancer cell lines, HCP5 silencing inhibited cell proliferation, migration, invasion, and promoted cell apoptosis via regulation of miR-299-3p/SMAD5 axis [ 36 ]. Meanwhile, HCP5 induced EMT processes via the miR-186-5p/WNT5A axis under hypoxia [ 9 ], and contributes to cisplatin resistance through miR-519d/HMGA1 and miR-128/HMGA2 axis [ 17 , 37 ].…”

Long non-coding RNA CERS6-AS1 plays a prognostic role in promoting the progression of gastric cancer

“…For example, lncRNA MAGI2-AS3 was previously proven to be an independent prognostic factor for the survival of gastric cancer patients [ 29 ]. The latest research also stated that lncRNA HCP5 may be a new and promising target for the treatment of gastric cancer [ 30 ]. Additionally, this study conducted a Kaplan-Meier curve analysis on the survival status of gastric cancer patients.…”

Long non-coding RNA titin-antisense RNA1 contributes to growth and metastasis of cholangiocarcinoma by suppressing microRNA-513a-5p to upregulate stratifin

“…Subsequently, the bottom chambers were filled with 500 μl RPMI-1640 medium containing 20% FBS. Following incubation for 24 h, cells passing through the membrane were immobilized with 4% paraformaldehyde, stained with 0.1% crystal violet, and visualized by microscopy [ 23 ].…”

“…CCLP and HCCC-9810 cells were co-transfected with the reporter vectors and miR-513a-5p mimic or negative control NC mimic using Lipofectamine 2000 (Invitrogen). Finally, luciferase activity was determined using the Dual-Luciferase Reporter Detection System (Promega, USA) in conformity with the supplier’s directions at 48 h after transfection [ 23 ].…”