CIZ1 aggravates gastric cancer progression via mediating FBXL19-AS1 and miR-339-3p.

Gastric cancer (GC) remains a prevalent malignancy with high morbidity and mortality. CDKN1A interacting zinc finger protein 1 (CIZ1) has been demonstrated to have oncogenic functions in the development of cancers. We detected CIZ1 expression via quantitative real-time PCR (RT-qPCR). The protein level of CIZ1 was measured through Western blot. We noticed that CIZ1 expression was markedly enhanced in GC cells. Furthermore, functional experiments including colony formation assay, EdU staining assay, transwell assay, TUNEL staining assay and flow cytometry analysis uncovered that CIZ1 silencing attenuated cell malignant phenotypes in GC. Through bioinformatics tools and mechanism assays, we explored the up-stream mechanism of CIZ1 and determined that CIZ1 was modulated by FBXL19 antisense RNA 1 (FBXL19-AS1) and microRNA-339-3p (miR-339-3p). Additionally, miR-339-3p exerted a negative role on GC development in vitro, and FBXL19-AS1 depletion also had the inhibitory impacts on the progression of GC in vitro. Eventually, the finding that CIZ1 overexpression reversed the effects of FBXL19-AS1 silencing on GC development was validated by rescue assays. In a word, CIZ1 functioned as a tumor promoter in GC, indicating that CIZ1 might be a promising target for GC treatment.

hsa_circ_0020378 regulating miR-339-3p/COL1A1 promotes osteosarcoma progression.

Osteosarcoma is a malignant orthopedic tumor that is frequently diagnosed in the pediatric population. Several studies have summarized the functions of circular RNAs (circRNAs) in the progression of osteosarcoma. This study aimed to investigate a novel circRNA, hsa_circ_0020378 (circ_0020378), and elucidate its functions and underlying mechanisms during osteosarcoma progression. The expression levels of circ_0020378, miR-339-3p, and COL1A1 in osteosarcoma cells and tissues were determined using RT-qPCR or Western blotting. CCK8, transwell migration, colony formation, and xenograft experiments were performed to assess the malignancy of osteosarcoma cells. Luciferase and RNA immunoprecipitation (RIP) experiments were employed to validate the interactions of miR-339-3p with circ_0020378 and COL1A1 3'UTR. Osteosarcoma cells and tissues showed significant upregulation of circ_0020378 and COL1A1 and downregulation of miR-339-3p. Silencing circ_0020378 in osteosarcoma cells inhibited their proliferation, colony formation, and migration. The inhibitive influence of circ_0020378 silencing during osteosarcoma tumorigenesis in vitro was verified in vivo. Circ_0020378 sponged miR-339-3p which targeted COL1A1 3'UTR. Circ _0020378 silencing disrupted the tumor-promoting effect of the miR-339-3p inhibitor in osteosarcoma cells. Furthermore, miR-339-3p inhibitor attenuated the suppressive effect of COL1A1 downregulation on malignant osteosarcoma cell phenotypes. Circ_0020378 stimulates osteosarcoma progression by downregulating miR-339-3p/COL1A1 expression. These findings provide a theoretical basis for the discovery of novel osteosarcoma targets.

KCNQ1OT1 promotes retinoblastoma progression by targeting miR-339-3p that suppresses KIF23.

BACKGROUND: Long noncoding RNAs (lncRNAs) are involved in tumor formation and development. KCNQ1OT1 regulates the malignant proliferation of retinoblastoma (RB), but the specific mechanism remains to be further investigated. METHODS: The KCNQ1OT1, miR-339-3p and KIF23 expression levels in RB were detected by qRT-PCR and western blotting. The cell viability, proliferation, migration ability and caspase-3 activity of RB cells were evaluated by CCK-8, BrdU, transwell and caspase-3 activity analysis. Western blot was used to detect the Bax and Bcl-2 protein expression in RB cells. The binding relationship between KCNQ1OT1, miR-339-3p and KIF23 was detected by luciferase, RIP and RNA pull-down assay. RESULTS: KCNQ1OT1 and KIF23 were up-regulated frequently in RB, and miR-339-3p was down-regulated. Functional studies showed that downregulation of KCNQ1OT1 or KIF23 inhibited the survival and migration of RB cells, and facilitated apoptosis. Interference with miR-339-3p showed the opposite effect. Mechanisms suggested that KCNQ1OT1 exited its oncogenic activity by positively regulating the expression of KIF23 and sponging miR-339-3p. CONCLUSION: KCNQ1OT1/miR-339-3p/KIF23 may be a new biomarker for the diagnosis and treatment of RB.

miR-339-3p promotes AT1-AA-induced vascular inflammation by upregulating NFATc3 protein expression in vascular smooth muscle cells.

Vascular inflammation induced by angiotensin II-1 receptor autoantibody (AT1-AA) is involved in the occurrence and development of various cardiovascular diseases. miR-339-3p is closely related to the degree of vasodilation of aortic aneurysm and is also involved in the occurrence and development of acute pancreatitis. However, it is still unclear whether miR-339-3p influences AT1-AA-induced vascular inflammation. In this study, the role and mechanism of miR-339-3p in AT1-AA-induced vascular inflammation are studied. RT-PCR detection shows that the miR-339-3p levels in the thoracic aorta and serum exosomes of AT1-AA-positive rats are significantly increased. The miRwalk database predicts the mRNAs that miR-339-3p can bind to their 5'UTR. Subsequently, it is found that the number of genes contained in the T cell receptor pathway is high through KEGG analysis, and NFATc3 among them can promote the secretion of various inflammatory cytokines. AT1-AA-induced upregulation of miR-339-3p expression in vascular smooth muscle cells (VSMCs) can lead to a significant increase in NFATc3 protein level and promote vascular inflammation. Inhibition of miR-339-3p with antagomir-339-3p can significantly reverse AT1-AA-induced high expressions of IL-6, IL-1ß and TNF-α proteins in rat thoracic aorta and VSMCs. That is, AT1-AA can upregulate the expression of miR-339-3p in VSMCs, and the increased miR-339-3p targets the 5'UTR of NFATc3 mRNA to increase the protein level of NFATc3, thereby aggravating the occurrence of vascular inflammation. These findings provide new experimental evidence for the involvement of miRNAs in regulating vascular inflammatory diseases.

Circ_0001821 Potentiates Cell Growth, Metastasis, and Stemness in Colorectal Cancer by Regulating miR-339-3p/CST1.

Dysregulation of circRNAs is associated with human cancer progression, but its expression pattern and function in cancer are not fully understood. The aim of our study is to investigate the effects of circ_0001821 on colorectal cancer (CRC) proliferation, migration, invasion, and stemness. The expression level of circ_0001821 in CRC tissues and cells was detected by quantitative real-time PCR. The effects of circ_0001821 silencing on the proliferation, migration, invasion, and stemness of CRC cells were examined by cell counting kit-8, 5-Ethynyl-2'-deoxyuridine, transwell, sphere formation, and western blot assays. Bioinformatics and dual-luciferase reporter gene assay were used to verify the relationship between circ_0001821 and miR-339-3p, miR-339-3p and CST1 in CRC cells. Circ_0001821 was highly expressed in CRC tissues and cells. Circ_0001821 silencing inhibited the proliferation, migration, invasion, and stemness of CRC cells, and transfection of miR-339-3p inhibitor partly attenuated this effect. In addition, circ_0001821 can bind miR-339-3p to regulate CST1 expression. Circ_0001821 silencing also curbed tumor growth in vivo. Circ_0001821 promoted CRC cell proliferation, migration, invasion, and stemness by regulating the miR-339-3p/CST1 axis.

Silencing LINC00665 inhibits cutaneous melanoma in vitro progression and induces apoptosis via the miR-339-3p/TUBB.

BACKGROUND: LncRNAs are closely related to cutaneous melanoma (CM) tumorigenesis and metastasis, and it can affect the progression of CM by regulating cell proliferation, migration, invasion, apoptosis, and other cellular mechanisms. This study investigated the role of LINC00665 in CM. METHODS: Expressions of LINC00665, miR-339-3p, and tubulin beta chain (TUBB) in CM cells were analyzed by qRT-PCR and/or Western blot. The LINC00665/miR-339-3p/TUBB targeting network was predicted by bioinformatics tools, screened out by Venn diagrams and analyzed by Pearson's correlation coefficients, followed by validation via dual-luciferase reporter assay and/or pull-down assay. Transfection of siLINC00665 or miR-339-3p inhibitor/mimic was conducted with CM cells whose viability, proliferation, migration, invasion, cell cycle progression, and apoptosis were measured by CCK-8 assay, colony formation assay, wound healing assay, Transwell assay, and flow cytometry. The associations of TUBB with tumor biological characteristics and other proteins were analyzed by CanserSEA and String, respectively. RESULTS: High-expressed LINC00665 was detected in CM cells. Silencing LINC00665 decreased CM cell viability; inhibited colony formation, cell cycle progression, migration and invasion; enhanced apoptosis; and upregulated miR-339-3p. LINC00665 targeted miR-339-3p which targeted TUBB. MiR-339-3p upregulation induced effects similar to the LINC00665-silencing-induced effects and could downregulate TUBB, which was associated with malignant behaviors and related to other five proteins. MiR-339-3p downregulation induced the opposite effects of what miR-339-3p upregulation induced, and the miR-339-3p downregulation-induced effects could be reversed by LINC00665 silencing. CONCLUSION: Silencing LINC00665 inhibits in vitro CM progression and induces apoptosis via the miR-339-3p/TUBB axis.

miR-339-3p inhibits cell growth and epithelial-mesenchymal transition in nasopharyngeal carcinoma by modulating the KAT6A/TRIM24 axis.

OBJECTIVE: To explore the effect and mechanism of the miR-339-3p/KAT6A/TRIM24 axis in nasopharyngeal carcinoma (NPC) cell growth and epithelial-mesenchymal transition (EMT) progression. METHODS: CNE2 and 5-8F NPC cell lines were transfected with miR-339-3p-mimic or sh-KAT6A alone or co-transfected with miR-339-3p-mimic and oe-KAT6A. The expression levels of miR-339-3p, KAT6A, TRIM24, and EMT-related proteins were assessed, in addition to cell biological behaviors. Then, the relationship between miR-339-3p and KAT6A was predicted and validated. The correlations between miR-339-3p and KAT6A or between KAT6A and TRIM24 were analyzed by Pearson coefficient and the enrichment of H3K23ac in TRIM24 promoter region was measured by chromatin immunoprecipitation. RESULTS: miR-339-3p was downregulated, but KAT6A and TRIM24 were highly expressed in NPC cells and tissues. Upregulated miR-339-3p or downregulated KAT6A could inhibit the growth and EMT of NPC cells. Further experiments showed that miR-339-3p regulated NPC cell growth and EMT by mediating KAT6A in a targeted fashion. KAT6A was positively correlated with TRIM24, and the enrichment of H3K23ac was much higher in NPC tissues. miR-339-3p suppressed the growth and EMT of NPC cells by the KAT6A/TRIM24 axis. In a xenograft study, miR-339-3p overexpression inhibited NPC tumor growth in vivo. CONCLUSION: Conclusively, miR-339-3p inhibited the growth and EMT of NPC cells via the KAT6A/TRIM24 axis.

Long Non-Coding NONRATG001910.2 Promotes the Proliferation of Rat Mesangial Cell Line HBZY-1 Through the miR-339-3p/CTNNB1 Axis.

Chronic glomerulonephritis (CGN) is one of the leading causes of end-stage renal disease (ESRD). A growing body of literature emphasizes the important role of long non-coding RNAs (lncRNAs) in the development and progression of the disease. However, the function of NONRATG001910.2 in the development of CGN was not well understood. This research aimed to investigate the effect of NONRATG001910.2 on CGN and revealed its potential molecular mechanisms. In this work, the expression of NONRATG001910.2 was detected by quantitative real-time polymerase chain reaction (qRT-RCR) in cell lines. We found that NONRATG001910.2 was significantly up-regulated in lipopolysaccharide (LPS) induced cells. High NONRATG001910.2 levels were associated with the development of CGN. In addition, NONRATG001910.2 knockdown inhibited cell proliferation and cell cycle. At the same time, we found that up-regulation of microRNA-339-3p (miR-339-3p) abrogated the biological roles of NONRATG001910.2 up-regulation. Moreover, the knockdown of CTNNB1 can upregulate miR-339-3p expression, thereby inhibiting cell proliferation. In conclusion, these results demonstrated that NONRATG001910.2 in LPS-stimulated rat mesangial cell line HBZY-1 (HBZY-1) by targeting miR-339-3p, which subsequently promotes the expression of CTNNB1, and suggested that NONRATG001910.2 may be a potential biomarker.

LINC00467 facilitates the proliferation, migration and invasion of glioma via promoting the expression of inositol hexakisphosphate kinase 2 by binding to miR-339-3p.

Our previous studies indicate that long noncoding RNA (lncRNA) LINC00467 can act as an oncogene to participate in the malignant progression of glioma, but the underlying molecular mechanism remains to be studied further. This study aimed to explore the biological role of the LINC00467/miR-339-3p/ inositol hexakisphosphate kinase 2 (IP6K2) regulatory axis in glioma. The Cancer Genome Atlas (TCGA), Oncomine databases and reverse transcription­quantitative PCR (RT­qPCR) were used to analyze IP6K2 expression in glioma. RT-PCR, EdU and transwell assays were conducted to observe the effect of IP6K2 on glioma cell proliferation, migration and invasion. Using bioinformatics analysis, RT-PCR, and dual luciferase reporter gene assay, the potential role of the LINC00467/miR-339-3p/IP6K2 regulatory axis in glioma was verified. The results showed that IP6K2 was up-regulated in glioma tissues and cell lines. Moreover, the expression level of IP6K2 was correlated with the clinical features of glioma patients. In vitro and in vivo experiments indicated that IP6K2 overexpression could promote the proliferation, migration, and invasion of glioma cells. Further bioinformatics analysis and in vitro assays revealed that LINC00467 could promote IP6K2 expression by binding to miR-339-3p and promote the malignant progression of glioma. Overall, LINC00467 could upregulate IP6K2 by binding to miR-339-3p and promote the proliferation, migration, and invasion of glioma cells. The LINC00467/miR-339-3p/IP6K2 regulatory axis might be a potential therapeutic target for glioma.

CircDOCK1 promotes the tumorigenesis and cisplatin resistance of osteogenic sarcoma via the miR-339-3p/IGF1R axis.

BACKGROUND: Circular RNAs (circRNAs), a class of noncoding RNAs (ncRNAs), may modulate gene expression by binding to miRNAs. Additionally, recent studies show that circRNAs participate in some pathological processes. However, there is a large gap in the knowledge about circDOCK1 expression and its biological functions in osteogenic sarcoma (OS). METHODS: Differentially expressed circRNAs in OS cell lines and tissues were identified by circRNA microarray analysis and quantitative real-time PCR (qRT-PCR). To explore the actions of circDOCK1 in vivo and in vitro, circDOCK1 was knocked down or overexpressed. To assess the binding and regulatory associations among miR-339-3p, circDOCK1 and IGF1R, we performed rescue experiments, RNA immunoprecipitation (RIP), RNA pulldown assays and dual-luciferase assays. Moreover, we performed apoptosis assays to reveal the regulatory effects of the circDOCK1/miR-339-3p/IGF1R axis on cisplatin sensitivity. RESULTS: CircDOCK1 expression remained stable in the cytoplasm and was higher in OS tissues and cells than in the corresponding controls. Overexpression of circDOCK1 increased oncogenicity in vivo and malignant transformation in vitro. In the U2OS and MG63 cell lines, circDOCK1 modulated tumor progression by regulating IGF1R through sponging of miR-339-3p. Additionally, in the U2OS/DDP and MG63/DDP cell lines, cisplatin sensitivity was regulated by circDOCK1 via the miR-339-3p/IGF1R axis. CONCLUSIONS: CircDOCK1 can promote progression and regulate cisplatin sensitivity in OS via the miR-339-3p/IGF1R axis. Thus, the circDOCK1/miR-339-3p/IGF1R axis may be a key mechanism and therapeutic target in OS.

LINC00467 knockdown repressed cell proliferation but stimulated cell apoptosis in glioblastoma via miR-339-3p/IP6K2 axis.

BACKGROUND: Glioma is considered to be one of the most common and lethal malignant brain tumors, accounting for 40% to 50% of brain tumors. Long non-coding RNAs (lncRNAs) have been widely proved to play an irreplaceable role in the tumorigenesis and progression. Nevertheless, the role of LINC00467 in glioblastoma remained unclear. AIM: The current study was aimed to explore the functional mechanism of LINC00467 in glioblastoma. METHODS: The expression of LINC00467/miR-339-3p/IP6K2 glioblastoma tissues and cells was evaluated by RT-qPCR. The protein expression of genes (cleaved PARP, PARP, cleaved caspase 3, caspase 3, Bax, Bcl-2 and IP6K2) was measured by western blot assay. Then role of LINC00467 was demonstrated by EdU, colony formation, flow cytometry and TUNEL assays. The relationship between miR-339-3p and LINC00467/IP6K2 was validated by RNA pull down and luciferase reporter assays. RESULTS: The expression of LINC00467 was upregulated in glioblastoma tissues and cells. LINC00467 knockdown suppressed cell proliferation but activated cell apoptosis. Further, LINC00467 high expression was associated with shorter overall survival rate in glioblastoma patients. Further, LINC00467 could bind with miR-339-3p, and IP6K2 was targeted by miR-339-3p. IP6K2 expression was regulated by LINC00467/miR-339-3p in a ceRNA pattern. Moreover, LINC00467 could regulate the development of glioblastoma via miR-339-3p/IP6K2 axis. CONCLUSIONS: LINC00467 knockdown repressed cell proliferation but stimulated cell apoptosis in glioblastoma via miR-339-3p/IP6K2 axis, which may enlighten to find a novel therapeutic tactic for glioblastoma patients.

miR-339-3p regulated acute pancreatitis induced by caerulein through targeting TNF receptor-associated factor 3 in AR42J cells.

Acute pancreatitis (AP) is an inflammatory disease with high morbidity and mortality. The regulation mechanism of miRNA is involved in the production and development of various diseases, but the regulation mechanism of miRNA in AP is still not fully elucidated. The expression of miR-339-3p was detected using quantitative real-time PCR. The levels of TNF-α, IL-1ß, and IL-6 were detected using enzyme-linked immunosorbent assay. Cell apoptosis was measured using flow cytometry. The protein expressions of TNF receptor-associated factor 3 (TRAF3), Bcl-2, C-caspase 3, Bax, p-p38, and p38 were measured using western blot. Luciferase reporter assay and RNA immunoprecipitation assay were applied to ensure that miR-399-3p targeted TRAF3. Caerulein promoted the expression of TNF-α, IL-1ß, and IL-6, enhanced the expression of C-caspase 3 and Bax while inhibited Bcl-2 protein expression. Meanwhile, caerulein also reduced the expression of miR-339-3p and induced the expression of TRAF3 in rat pancreatic acinar cells. miR-399-3p transfection inhibited the levels of TNF-α, IL-1ß, and IL-6 and C-caspase 3 and Bax protein expression as well as suppressed cell apoptosis, while increased Bcl-2 protein expression in caerulein-induced AP. TRAF3 has been verified as a target of miR-339-3p. Interestingly, the reduction of miR-399-3p inhibited the p38 pathway, which was impaired by the upregulation of TRAF3. In addition, the suppression effects of miR-339-3p on cell inflammation and apoptosis in caerulein-induced AP were reversed by enhancing TRAF3 expression. In this study, in vitro model of AP was characterized by strong inflammation and cell apoptosis. We have first demonstrated the regulatory network of miR-339-3p and TRAF3. Overexpression of miR-339-3p inhibited cell inflammation and cell apoptosis in caerulein-induced AP through modulating TRAF3 expression via the p38 pathway, providing a new therapeutic target in the treatment of AP.

miR-339-3p inhibits proliferation and metastasis of colorectal cancer.

MicroRNAs (miRNAs) serve important roles in regulating cancer cell proliferation and metastasis. The same hairpin RNA structure may produce mature products from each strand, termed miR-5p and miR-3p, which can bind different mRNAs. Previously, the present authors reported that miR-339-5p could inhibit cell proliferation and migration by targeting the 3'-untranslated region (3'-UTR) of PRL-1 mRNA. The present study analyzed the expression, function and preliminary regulatory mechanism of miR-339-3p in colorectal cancer (CRC). The results of reverse transcription-quantitative polymerase chain reaction analysis demonstrated that miR-339-3p is downregulated in CRC specimens and highly invasive cell lines. Furthermore, the low-level expression of miR-339-3p was significantly associated with lymph node metastasis in patients with CRC; however, reduced miR-339-3p expression was not associated with age, gender or the differentiation status of the tumor. Overexpression of miR-339-3p was sufficient to suppress tumor growth and metastasis in vitro. In addition, the present study demonstrated that unlike miR-339-5p, PRL-1 expression was not regulated by miR-339-3p. The findings of the present study indicate that miR-339-5p and miR-339-3p may target different mRNA. The target gene of miR-339-3p requires future identification.