ABSTRACT
BACKGROUND: Long noncoding RNA (lncRNA) plays critical roles in both tumor-suppressive and oncogenic pathways in the pathological development and prognosis of cancers. AIMS: This study aimed to explore the expression of lncRNA AFAP1-AS1 and its function in gastric cancer (GC). METHODS: The expression of AFAP1-AS1 was detected in GC tissues and GC cells by quantitative real-time reverse-transcription PCR. A small interfering RNA (siRNA) that targeted AFAP1-AS1 was transfected into cells to inhibit the expression of AFAP1-AS1. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay and colony formation assay were performed to examine the cell proliferation of SGC7901 cell transfected with si-AFAP1-AS1. Cell apoptosis was detected by flow cytometry. The protein level of cleaved PARP, Caspase 3, Caspase 9, Caspase 8, Bcl-2, Bax, p-AKT, total-AKT, and PTEN were detected by Western blot. RESULTS: AFAP1-AS1 was up-regulated in GC tissues and GC cells. AFAP1-AS1 knockdown suppressed cell viability of SGC7901 transfected with si-AFAP1-AS1. The number of apoptotic SGC7901 cell transfected with si-AFAP1-AS1 was increased by 3.4-fold comparing to that of control. The protein level of cleaved PARP, Caspase 3, and Caspase 9 were increased in SGC7901 transfected with si-AFAP1-AS1, as well as the expression of Bax. The protein level of Bcl-2 was decreased. AFAP1-AS1 knockdown decreased the protein level of p-AKT and increased the expression of PTEN in SGC7901 cells. CONCLUSIONS: AFAP1-AS1 was up-regulated in GC cells and regulated the gastric cancer cell proliferation and apoptosis via PTEN/p-AKT pathway.
Subject(s)
Apoptosis/genetics , Cell Proliferation/genetics , RNA, Long Noncoding/physiology , Signal Transduction/genetics , Stomach Neoplasms/genetics , Stomach Neoplasms/pathology , Carcinogenesis/genetics , Cell Line, Tumor , Gene Knockdown Techniques , Humans , PTEN Phosphohydrolase/metabolism , Proto-Oncogene Proteins c-akt/metabolism , RNA, Small Interfering/metabolism , Up-RegulationABSTRACT
Increasing evidences indicate that dys-regulation of MicroRNAs contributes to hepatocellular carcinoma. However, the roles of miR-485-5p in HCC are still largely unexplored. In the present study, our quantitative real-time PCR analysis found that miR-485-5p was significantly down-regulated in 50 pairs of human HCC tissues. Moreover, the reduced expression of miR-485-5p was significantly correlated with larger tumor size and more tumor number in patients with HCC. In vitro studies further showed that overexpression of miR-485-5p mimics could inhibit, while its antisense oligos promote cell proliferation and invasion. Results from the dual-luciferase reporter gene assays and western blot further showed that stanniocalcin 2 was a direct target of miR-485-5p. Therefore, our data suggest a novel role for miR-485-5p in the regulation of HCC progression.
