RETRACTED ARTICLE: MicroRNA-330-3p promotes cell invasion and metastasis in non-small cell lung cancer through GRIA3 by activating MAPK/ERK signaling pathway

Abstract

BackgroundBrain metastasis (BM) is associated with poor prognosis in patients with non-small cell lung cancer (NSCLC). Recent studies demonstrated that microRNA-330-3p (miR-330-3p) was involved in NSCLC brain metastasis (BM). However, the exact parts played by miR-330-3p in BM of NSCLC remain unknown. Discovery and development of biomarkers and elucidation of the mechanism underlying BM in NSCLC is critical for effective prophylactic interventions. Here, we evaluated the expression and biological effects of miR-330-3p in NSCLC cells and explored the underlying mechanism of miR-330-3p in promoting cell migration and invasion in NSCLC.MethodsStable over-expression and knockdown of miR-330-3p in NSCLC cells was constructed with lentivirus. Expression levels of miR-330-3p in NSCLC cells were quantified by quantitive real-time PCR (qRT-PCR). The effects of miR-330-3p on NSCLC cells were investigated using assays of cell viability, migration, invasion, cell cycle, apoptosis, western blotting, immunohistochemical, and immunofluorescence staining. A xenograft nude mouse model and in situ brain metastasis model were used to observe tumor growth and brain metastasis. The potential target of miR-330-3p in NSCLC cells was explored using the luciferase reporter assay, qRT-PCR, and western blotting. The miR-330-3p targets were identified using bioinformatics analysis and verified by luciferase reporter assay. The correlation between GRIA3 and DNA methyltransferase (DNMT) 1 and DNMT3A was tested by RT-PCR, western blotting, and co-immunoprecipitation (IP).ResultsmiR-330-3p was significantly up-regulated in NSCLC cell lines. MTT assay, transwell migration, and invasion assays showed that miR-330-3p promoted the growth, migration, and invasion of NSCLC cells in vitro and induced tumor growth and metastasis in vivo. Luciferase reporter assays showed that GRIA3 was a target of miR-330-3p. qRT-PCR and western blotting exhibited that miR-330-3p promoted the growth, invasion, and migration of NSCLC cells by activating mitogen-activated protein kinase (MAPK)/extracellular-regulated protein kinases (ERK) signaling pathway. Furthermore, miR-330-3p up-regulated the total DNA methylation in NSCLC cells, and co-IP-demonstrated GRIA3 was directly related with DNMT1 and DNMT3A.ConclusionsmiR-330-3p promoted the progression of NSCLC and might be a potential target for the further research of NSCLC brain metastasis.Electronic supplementary materialThe online version of this article (doi:10.1186/s13045-017-0493-0) contains supplementary material, which is available to authorized users.

Expression of Glutamate Receptor Subtype 3 Is Epigenetically Regulated in Podocytes under Diabetic Conditions

“…Pathway analysis of the total of 339 genes showed that altered genes are largely involved in cell division and cell cycle regulation (Table 1). For two hypermethylated genes in our analysis, ESX1, a homeodomain protein, was reported to regulate cell cycle progression and transcription [24-26], and GRIA3 has been shown to promote proliferation in nonneuronal cells [27, 28]. As other glutamate receptors, such as NMDA receptor, have an important function in podocyte biology and glomerular diseases, we selected GRIA3 for further analysis.…”

“…Kidney sections and isolated glomeruli were obtained from control (control-eNOS –/– ) and diabetic (STZ-eNOS –/– ) mice as described previously [27]. a Real-time PCR of Gria3 in isolated glomeruli is shown as a relative fold change to control mice.…”

“…We now show that GRIA3 is epigenetically regulated in podocytes under HG conditions, and that GluR3, encoded by GRIA3 , confers protection against podocyte apoptosis in HG conditions. Recent reports indicate that GluR3 promotes cell growth through activation of MAPK pathway [27]. Therefore, GluR3 may be a survival factor for podocytes through activation of the MAPK pathway [34, 35].…”

Expression profile of SYNE3 and bioinformatic analysis of its prognostic value and functions in tumors

“…It was reported that miR-330-3p can target the gene of program cell death 4 (PDCD4) to promote tumor [17]. MiR-330-3p is also related with metastasis and invasion of lung cancer, and glutamate receptor 3 (GRIA3) [18] and human manganese superoxide dismutase 2 (hSOD2) [19] are its two target genes. Researches found miR-330-3p is correlated with bad prognosis in breast cancer as well [20], in which miR-330-3p can target Collagen and Calcium Binding EGF Domains 1 (CCBE1) to promote cancer metastasis [21].…”

<p>Osteopontin Mediates Cetuximab Resistance via the MAPK Pathway in NSCLC Cells</p>

“…It is well known that the MAPK pathway participates in regulating the invasion and metastasis of NSCLC; 26 thus, we explored the potential effects of OPN on the MAPK pathway. Western blotting showed that OPN overexpression increased p-MEK and p-ERK in A549 cells, and silencing of OPN had the opposite results ( Figure 3A and B).…”