MicroRNA‑125 inhibits RKO colorectal cancer cell growth by targeting VEGF

Abstract

Colorectal cancer (CRC) is one of the major types of cancer and causes of mortality worldwide, and it remains the third most common cause of cancer‑associated mortality worldwide. MicroRNAs (miRNAs) are a class of small RNAs, which have been shown to be associated with CRC. In the present study, an MTT assay and proliferating cell nuclear antigen (PCNA) protein examination assay were performed to detect RKO cell viability. Hoechst staining, and caspase‑3 activity and BrdU incorporation assays were performed to detect RKO cell apoptosis, respectively. Reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) and western blot analyses were used to analyze the expression of cyclooxygenase‑2 (COX‑2). Western blot analysis was also used to analyze the expression of vascular endothelial growth factor (VEGF) and mitogen‑activated protein kinase (MAPK) signal molecules, including extracellular signal‑regulated kinase (ERK), p38 and c‑Jun N‑terminal kinase (JNK). The target genes of miR-125 were predicted using a double luciferase reporter gene assay. The results of the MTT assay showed that RKO cell viability was decreased by an miRNA-125 mimic and increased by the miRNA-125 inhibitor. The RKO cell viability was significantly correlated with the expression of PCNA. The migration of RKO cells was significantly downregulated in the miR-125 mimics‑transfected cells and upregulated in the miRNA-125 inhibitor‑transfected cells. The results of Hoechst staining and the caspase‑3 activity and BrdU incorporation assays showed that RKO cell apoptosis was increased following miRNA-125 mimic transfection and decreased following miRNA-125 inhibitor transfection. The results of the RT‑qPCR and western blot analysis showed that the expression of COX‑2 was increased in the miR-125 mimic‑transfected cells and decreased in the miR-125 inhibitor‑transfected cells. Using an online miRNA target prediction database, the double luciferase reporter gene assay showed that miR‑125 targeted and inhibited the expression of VEGF through target sites located in the 3' untranslated region of VEGF mRNA. In conclusion, the abnormal expression of miR‑125 was found to be closely associated with CRC. Therefore, miR‑125 may be a novel therapeutic target for CRC.

MicroRNA‑125a‑5p regulates liver cancer cell growth, migration and invasion and EMT by targeting HAX1

“…In laryngeal cancer, Zhang et al (52) demonstrated that downregulated miR-206 promoted the proliferation and invasion of cancer cells by modulating VEGF expression. Wu et al (53) suggested that miR-125 suppressed cell growth of RKO colorectal cancer cells via targeting VEGF. It was reported that the EMT process is involved in the migration and invasion of cancer cells and plays a critical role in cancer metastasis (54,55).…”

Steroid 5α-Reductase Type I Induces Cell Viability and Migration via Nuclear Factor-κB/Vascular Endothelial Growth Factor Signaling Pathway in Colorectal Cancer

“…Cell cycle and apoptosis were analyzed by using flow cytometry, of which the protocol was reported in the literature ( 31 – 33 ). For the cell cycle, around 1 × 10 6 cells were washed by cold PBS and fixed in 70% ethanol at −20°C overnight.…”

Study of the Association between microRNA (miR-25T>C, miR-32C>A, miR-125C>T, and miR-222G>T) Polymorphisms and the Risk of Recurrent Pregnancy Loss in Korean Women

“…Many studies have revealed that miRNA polymorphisms can affect the levels of VEGF and VEGFRs, such as KDR, leading to the inhibition of the VEGF pathway and abnormal angiogenesis [6,34,35]. In particular, understanding the mechanisms underlying the miRNA-mediated regulation of the VEGF-VEGFR pathway during pregnancy and fetal development is critical, as angiogenesis plays key roles in establishing and maintaining pregnancy [31,36].…”