Downregulation of hsa_circTLK1 represses non-small cell lung cancer progression by regulating miR-876-3p/SRSF7 axis.

Background: This study clarified the expression of cicrTLK1 in non-small cell lung cancer (NSCLC) and explored its role in cancer growth, metastasis and immune escape, providing a potential molecular target and theoretical basis for NSCLC treatment. Methods: The expression levels of circTLK1, miR-876-3p and SRSF7 were determined by RT-qPCR assay. The localization of circTLK1 in NSCLC cells was determined by FISH assay. EdU and cell plate clone formation assay were applied to explore cell proliferation. Wound healing test and Transwell assay were applied to measure the migration and invasion ability. Cell apoptosis rate was detected by FCM assay. Western blotting assay was adopted to measure the protein expression of SRSF7. Dual-luciferase reporter gene assay was applied to assess the interaction between miR-876-3p and circTLK1, and between miR-876-3p and SRSF7. The ability of cirTLK1 to regulate tumor formation in vivo was examined by tumor transplantation experiments in nude mice. Results: The relative expression of circTLK1 was increased in NSCLC cell lines. Knockdown of circTLK1 prohibited the proliferation, migration, and invasion, and promoted apoptosis rate, but miR-876-3p inhibitor reversed the effects of circTLK1 knockdown. In addition, silencing of circTLK1 overtly restrained the growth of transplanted tumors in vivo, and inhibited immune escape. In addition, circTLK1 interacted with miR-876-3p, and SRSF7 was concluded to be the target gene of miR-876-3p. Conclusion: In this study, we researched the inhibitory effect of circTLK1knockdown on NSCLC progression and immune escape, and further elucidated the potential regulatory mechanism of circTLK1/miR876-3p/SRSF7 axis.

Differential effect of cancer-associated fibroblast-derived extracellular vesicles on cisplatin resistance in oral squamous cell carcinoma via miR-876-3p.

Rationale: Platinum-based chemotherapy is commonly used for treating solid tumors, but drug resistance often limits its effectiveness. Cancer-associated fibroblast (CAF)-derived extracellular vesicle (EV), which carry various miRNAs, have been implicated in chemotherapy resistance. However, the molecular mechanism through which CAFs modulate cisplatin resistance in oral squamous cell carcinoma (OSCC) is not well understood. We employed two distinct primary CAF types with differential impacts on cancer progression: CAF-P, representing a more aggressive cancer-promoting category, and CAF-D, characterized by properties that moderately delay cancer progression. Consequently, we sought to investigate whether the two CAF types differentially affect cisplatin sensitivity and the underlying molecular mechanism. Methods: The secretion profile was examined by utilizing an antibody microarray with conditioned medium obtained from the co-culture of OSCC cells and two types of primary CAFs. The effect of CAF-dependent factors on cisplatin resistance was investigated by utilizing conditioned media (CM) and extracellular vesicle (EVs) derived from CAFs. The impacts of candidate genes were confirmed using gain- and loss-of-function analyses in spheroids and organoids, and a mouse xenograft. Lastly, we compared the expression pattern of the candidate genes in tissues from OSCC patients exhibiting different responses to cisplatin. Results: When OSCC cells were cultured with conditioned media (CM) from the two different CAF groups, cisplatin resistance increased only under CAF-P CM. OSCC cells specifically expressed insulin-like growth factor binding protein 3 (IGFBP3) after co-culture with CAF-D. Meanwhile, IGFBP3-knockdown OSCC cells acquired cisplatin resistance in CAF-D CM. IGFBP3 expression was promoted by GATA-binding protein 1 (GATA1), a transcription factor targeted by miR-876-3p, which was enriched only in CAF-P-derived EV. Treatment with CAF-P EV carrying miR-876-3p antagomir decreased cisplatin resistance compared to control miRNA-carrying CAF-P EV. On comparing the staining intensity between cisplatin-sensitive and -insensitive tissues from OSCC patients, there was a positive correlation between IGFBP3 and GATA1 expression and cisplatin sensitivity in OSCC tissues from patients. Conclusion: These results provide insights for overcoming cisplatin resistance, especially concerning EVs within the tumor microenvironment. Furthermore, it is anticipated that the expression levels of GATA1 and miR-876-3p, along with IGFBP3, could aid in the prediction of cisplatin resistance.

A Novel circ_0104345/miR-876-3p/ZBTB20 Axis Regulates the Proliferation, Migration, Invasion, and Apoptosis of Breast Cancer Cells.

Breast cancer (BC) is one of the most common malignant tumors in women. CircRNA/miRNA/mRNA regulatory axes have been shown to be involved in the pathogenesis of BC. Here, we sought to analyze the functional mechanism of circ_0104345 in BC. Quantitative real-time polymerase chain reaction (qRT-PCR) was performed to detect the levels of circ_0104345, miR-876-3p and zinc finger and BTB domain containing 20 (ZBTB20) mRNA. Cell Counting Kit-8 (CCK8) and 5-ethynyl-2'-deoxyuridine (EdU) assays were used to test cell viability and proliferation, respectively. Cell migration was tested by wound healing assay, and cell invasion was examined by transwell assay. Tube formation ability was tested by angiogenesis assay. Flow cytometry was applied for cell apoptosis. Western blot assay was utilized to measure the protein expression. The relationship between miR-876-3p and circ_0104345 or ZBTB20 was identified by dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay. Xenografts in mice were conducted to analyze the effect of sh-circ_0104345 on tumor growth in vivo. Circ_0104345 and ZBTB20 were upregulated and miR-876-3p expression was decreased in BC. Circ_0104345 knockdown inhibited cell proliferation, migration, invasion, and enhanced cell apoptosis. MiR-876-3p was targeted by circ_0104345. MiR-876-3p depletion reversed the effects of circ_0104345 downregulation on the progression of BC cells. ZBTB20 was regulated by circ_0104345 through miR-876-3p. The effects of miR-876-3p on BC cell behaviors were restored by ZBTB20 increase. The results of in vivo experiments indicated that silencing of circ_0104345 blocked the growth of xenograft tumors. In this study, we demonstrated, for the first time, the crucial regulation of the new circ_0104345/miR-876-3p/ZBTB20 axis in the biological phenotypes of BC cells.

The expression of miR-876-3p based on a rat model of bronchopulmonary dysplasia induced by hyperoxia / 中国小儿急救医学

Objective:To establish a neonatal rat bronchopulmonary dysplasia(BPD) model induced by hyperoxia, to detect the expression of miR-876-3p in the lung tissue, and to analyze the role of miR-876-3p in the occurrence and development of BPD, so as to provide a theoretical basis for the pathogenesis, prevention and treatment of BPD.Methods:Eighty newborn SD rats were randomly divided into hyperoxia group(FiO 2 60%) and air group(FiO 2 21%). Lung tissue samples were taken on the 1st, 7th, 14th and 21st day after birth, the pathological changes of lung tissue were observed.Quantitative real-time PCR technique was used to detect the expression level of miR-876-3p. Results:Within 21 days after birth, with the prolongation of hyperoxia exposure time, the general growth of rats in hyperoxia group were lower than those in air group[14 d: (35.46±1.62) g vs.(37.08±1.25) g; 21 d: (51.92±1.83) g vs.(58.87±2.43) g]( P<0.05). On the 14th and 21st day after birth, the radial alveolar counts in lung tissue of rats in hyperoxia group were significantly reduced compared with those in air group( P<0.05). On the 7th, 14th and 21st day after birth, the alveolar septal thickness of rats in air group were lower than those in hyperoxia group( P<0.05). The expression level of miR-876-3p in hyperoxia group decreased gradually and was significantly lower on the 7th, 14th and 21st day compared with air group at the same time points[7 d: (14.97±1.13) vs.(16.64±0.89); 14 d: (11.92±0.71) vs.(16.85±0.79); 21 d: (11.39±0.79) vs.(17.52±1.17)], and the differences were all statistically significant( P all<0.01). Conclusion:In this study, a new BPD model of neonatal rats can be induced by hyperoxia and the expression level of miR-876-3p in this model is decreased.The differential expression level of miR-876-3p may play a role in the occurrence and development of BPD.

Hsa_circ_0043949 reinforces temozolomide resistance via upregulating oncogene ITGA1 axis in glioblastoma.

Temozolomide (TMZ) resistance limits its use in glioblastoma (GBM). Exosomes can carry circular RNAs (circRNAs) to regulate chemoresistance. To date, the role of exosomal hsa_circ_0043949 (circ_0043949) in GBM resistance to TMZ is unclear. Relative expression of circ_0043949 in clinical samples, GBM cell lines, and exosomes was detected by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The half-maximal inhibitory concentration (IC50) of TMZ, cell proliferation, apoptosis, invasion, and migration were analyzed via MTT, EdU, flow cytometry, transwell, and wound-healing assays. Relative protein levels were evaluated by western blotting. Target relationship was predicted by bioinformatics analysis and validated by dual-luciferase reporter and RNA pull-down assays. Exosomes were isolated by ultracentrifugation and verified by transmission electron microscopy, nanoparticle tracking analysis (NTA), and western blotting. The effect of exosomal circ_0043949 on TMZ resistance was validated by xenograft assay. Higher expression of circ_0043949 was gained in TMZ-resistant GBM samples and cells. Inhibition of circ_0043949 reduced TMZ resistance via decreasing IC50 of TMZ, repressing proliferation, invasion, migration, and inducing apoptosis in TMZ-resistant GBM cells. Circ_0043949 mediated integrinalpha1 (ITGA1) expression via function as a miR-876-3p sponge. Circ_0043949 was also upregulated in TMZ-resistant GBM cells-derived exosomes, and exosomal circ_0043949 increased the resistance of TMZ-resistant GBM cells to TMZ in xenograft models. TMZ-resistant GBM cells-derived exosomal circ_0043949 promoted TMZ resistance via upregulating ITGA1 expression via sequestering miR-876-3p, offering a potential target for the treatment of TMZ resistance in GBM.

CircNFIX knockdown inhibited AML tumorigenicity by the miR-876-3p/TRIM31 axis.

BACKGROUND: Acute myeloid leukemia (AML) is one of the most common malignant myeloid diseases in adults with a dismal prognosis. We aimed to explore the effects of circNFIX on the proliferation and apoptosis of AML cells. METHODS: The expressions of circNFIX, miR-876-3p and tripartite motif (TRIM) 31 in the bone marrow specimens of AML patients and AML cell lines were detected by qRT-PCR or western blot. Cell proliferation was evaluated by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and 5-ethynyl-29-deoxyuridine (EdU) assays. Cell cycle and apoptosis were analyzed by flow cytometry. Western blot was used to detect protein expression. The relationship between miR-876-3p and circNFIX or TRIM31 was identified by dual-luciferase reporter assay or RNA pull-down assay. RESULTS: The expression level of circNFIX was significantly increased in the bone marrow samples of AML patients and AML cells when compared with normal controls. CircNFIX silencing inhibited AML cell proliferation and promoted apoptosis. Inhibition of miR-876-3p reversed the effect of circNFIX knockdown on AML cell progression. In addition, circNFIX indirectly regulated TRIM31 through miR-876-3p. Further, TRIM31 overexpression counteracted the effect of circNFIX silencing on AML cell proliferation and apoptosis. CONCLUSION: CircNFIX knockdown could suppress the proliferation and induce the apoptosis of AML cells by targeting the miR-876-3p/TRIM31 axis.

Hsa_circ_0097922 promotes tamoxifen resistance and cell malignant behaviour of breast cancer cells by regulating ACTN4 expression via miR-876-3p.

An increasing number of findings have verified the critical roles of circular RNAs (circRNAs) in human cancers, and chemotherapy resistance is a poor prognostic factor for breast cancer (BC). This study is designed to explore the function of hsa_circ_0097922 in the tamoxifen resistance of breast cancer. Hsa_circ_0097922, microRNA-876-3p (miR-876-3p), and alpha-actinin 4 (ACTN4) level were detected by real-time quantitative polymerase chain reaction (RT-qPCR). Cell survival, proliferation, apoptosis, migration and invasion were detected by Cell Counting Kit-8 (CCK-8), 5-ethynyl-2'-deoxyuridine (EdU), and flow cytometry, wound healing and Transwell assays. Protein levels of proliferating cell nuclear antigen (PCNA), B-cell lymphoma-2 (Bcl-2), cleaved caspase 3, matrix metalloproteinase 9 (MMP9), and ACTN4 were determined using western blot assay. Using bioinformatics software, the binding between miR-876-3p and hsa_circ_0097922 or ACTN4 was predicted, followed by confirmation by RNA immunoprecipitation (RIP) and RNA pull-down assays. A xenograft tumour model in vivo analysed the biological role of hsa_circ_0097922 on BC tumour growth and drug resistance. Hsa_circ_0097922 and ACTN4 were increased, and miR-876-3p was decreased in tamoxifen resistance BC cells. Moreover, hsa_circ_0097922 knockdown can block BC cell malignant behaviour and tamoxifen resistance in vitro. Mechanically, hsa_circ_0097922 acted as a sponge of miR-876-3p to regulate ACTN4 expression. Hsa_circ_0097922 silencing increased the drug sensitivity of BC in vivo. Hsa_circ_0097922 might regulate BC cell malignant behaviour and tamoxifen resistance partly by regulating the miR-876-3p/ACTN4 axis, hinting at a promising therapeutic target for the BC treatment.

Circ_0016760 Serves as a Cancer Promoter in Non-small Cell Lung Cancer Through miR-876-3p/NOVA2 Axis.

Non-small cell lung cancer (NSCLC) is a serious threaten to human health globally. Circular RNAs (circRNAs) were testified to alter the progression of NSCLC. This work intended to investigate the functional role of circ_0016760 in NSCLC development and the potential mechanism. Expression of circ_0016760, microRNA (miR)-876-3p and NOVA alternative splicing regulator 2 (NOVA2) was determined via quantitative reverse transcription-PCT (qRT-PCR) or western blotting. Cell viability, clonogenicity and apoptosis were assessed by Cell Counting Kit-8 (CCK-8) assay, colony formation assay and flow cytometry, respectively. Transwell assay was performed to examine cell migration and invasion. Western blotting was also conducted to detect the levels of epithelial-to-mesenchymal transition (EMT)-related proteins. Role of circ_0016760 in vivo was evaluated via xenograft model assay. Moreover, the interaction between miR-876-3p and circ_0016760 or NOVA2 was verified by dual-luciferase reporter assay or RNA Immunoprecipitation (RIP) assay. Circ_0016760 and NOVA2 were upregulated, while miR-876-3p expression was decreased in NSCLC tissues and cells. Circ_0016760 depletion suppressed NSCLC cell proliferation and metastasis in vitro, as well as hampered tumor growth in vivo. Circ_0016760 acted as a sponge of miR-876-3p, and miR-876-3p could target NOVA2. Circ_0016760 might play vital roles in NSCLC by regulating miR-876-3p/NOVA2 axis. Circ_0016760 could promote the malignant development of NSCLC through miR-876-3p/NOVA2 axis, at least in part.

Circ_0062558 promotes growth, migration, and glutamine metabolism in triple-negative breast cancer by targeting the miR-876-3p/SLC1A5 axis.

BACKGROUND: Circular RNAs (circRNAs) have been reported to function as vital regulators in cancers, including triple-negative breast cancer (TNBC). This study aimed to explore the role of circ_0062558 in TNBC. METHODS: The real-time quantitative polymerase chain reaction (RT-qPCR) was conducted to quantify the expressions of circ_0062558, microRNA-876-3p (miR-876-3p), and solute carrier family 1 (neutral amino acid transporter), member 5 (SLC1A5) in TNBC tissues and cells. 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyl-2H-tetrazol-3-ium bromide (MTT), thymidine analog 5-ethynyl-2'-deoxyuridine (EdU), flow cytometry, wound healing, and Transwell assays were employed for cell phenotype analyses. Protein expression was tested by western blot analysis. Dual-luciferase reporter was used to confirm the association among circ_0062558, miR-876-3p, and SLC1A5 in TNBC. Xenograft experiments were performed to elucidate the function of circ_0062558 in vivo. RESULTS: TNBC tissues and cells showed the higher level of circ_0062558 when compared with control samples. Downregulation of circ_0062558 inhibited proliferation, migration, invasion, and glutamine metabolism, while enhanced apoptosis of TNBC cells, and silencing of circ_0062558 also inhibited the growth of tumor in vivo. MiR-876-3p was confirmed as a target of circ_0062558, and circ_0062558 knockdown repressed TNBC cell malignant behaviors by increasing miR-876-3p. Furthermore, miR-876-3p inhibited malignant behaviors of TNBC cells by down-regulating SLC1A5, a newly identified target of miR-876-3p. CONCLUSION: Circ_0062558 promoted TNBC progression by enhancing proliferation, survival, migration, invasion, and glutamine metabolism via miR-876-3p/SLC1A5 axis, which was helpful for understanding the carcinogenic roles of circ_0062558.

CircLNPEP promotes the progression of ovarian cancer through regulating miR-876-3p/ WNT5A axis.

CircRNA LNPEP has been shown to promote the development of hepatocellular carcinoma, but its function in ovarian cancer (OC) remains unclear. The Kaplan-Meier method was used to analyze the clinical significance of circLNPEP expression in OC patients. The stability of circLNPEP was detected by actinomycin D and RNase R treatment. The correlations between miR-876-3p and two genes (circLNPEP and WNT5A) were predicted by bioinformatics analysis and confirmed by dual-luciferase reporter assay. Expressions of circLNPEP, miR-876-3p, and WNT5A were determined by qRT-PCR and western blot. The effect of circLNPEP/miR-876-3p/WNT5A axis on viability, proliferation, migration, and invasion, and angiogenesis of cells was determined by cell function experiment and rescue experiment. Xenograft tumor mice were constructed for in vivo verification. Expressions of apoptosis, epithelial mesenchymal transition (EMT)-related genes, and CD34 were determined by qRT-PCR, western blot and immunohistochemistry. High level of circLNPEP was related to poor prognosis in OC. CircLNPEP was highly expressed in OC tissues and cell lines, mainly distributed in the cytoplasm, while miR-876-3p was the opposite. MiR-876-3p targeted and negatively correlated with circLNPEP and WNT5A. Sh-circLNPEP repressed cell viability, proliferation, migration, invasion, angiogenesis, and EMT but promoted apoptosis, which were related to its regulation of expression of EMT- and apoptosis-related genes, WNT5A, and CD34. The regulatory effect of sh-circLNPEP can be reversed by miR-876-3p inhibitor, and that of miR-876-3p inhibitor can be reversed by sh-WNT5A. CircLNPEP promoted cancer cell proliferation, EMT and angiogenesis, and inhibited apoptosis by regulating miR-876-3p/WNT5A axis.

miR-876-3p suppresses the progression of colon cancer and correlates the prognosis of patients.

BACKGROUND: miR-876-3p has been identified to be downregulated in colon cancer, implying the potential biological function in the progression and prognosis of colon cancer. The clinical significance and the biological function of miR-876-3p were investigated in this study to assess the potential of miR-876-3p in acting as a novel biomarker of the progression of colon cancer. METHODS: The expression of miR-876-3p in colon cancer was evaluated by RT-qPCR. The clinical significance of miR-876-3p was assessed by associated its expression level with the clinical features and prognosis of patients. The biological function of miR-876-3p was estimated by the CCK8 and Transwell assay in vitro. RESULTS: The significant downregulation of miR-876-3p was observed in colon cancer tissues and cells, which was closely associated with the lymph node metastasis status, TNM stage, and the perineural invasion of patients. miR-876-3p served as an independent indicator that was negatively associated with the prognosis of patients. In colon cancer cells, miR-876-3p showed significant inhibitory effects on cell proliferation, migration, and invasion, indicating its tumor suppressor role in the progression of colon cancer. CONCLUSION: miR-876-3p might be involved in colon cancer development, which provides a potential therapeutic target for colon cancer treatment.

Circ-RNF111 aggravates the malignancy of gastric cancer through miR-876-3p-dependent regulation of KLF12.

BACKGROUND: The aberrant expression of circular RNAs (circRNAs) plays vital roles in the advancement of human cancers, including gastric cancer (GC). In this study, the functions of circRNA ring finger protein 111 (circ-RNF111) in GC were investigated. METHODS: Quantitative real-time polymerase chain reaction (qRT-PCR) assay was performed for the levels of circ-RNF111, microRNA-876-3p (miR-876-3p) and krueppel-like factor 12 (KLF12) mRNA. RNase R assay was conducted for the feature of circ-RNF111. Cell Counting Kit-8 (CCK-8) assay, colony formation assay, wound-healing assay, and transwell assay were applied for cell viability, colony formation, migration, and invasion, respectively. Flow cytometry analysis was used to analyze cell apoptosis and cell cycle process. The glycolysis level was examined using specific commercial kits. Western blot assay was carried out to measure the protein levels of hexokinase 2 (HK-2) and KLF12. Dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay were employed to verify the combination between miR-876-3p and circ-RNF111 or KLF12. Murine xenograft model was constructed for the role of circ-RNF111 in vivo. Immunohistochemistry (IHC) was used for KLF12 level. RESULTS: Circ-RNF111 was higher expressed in GC tissues and cells than normal tissues and cells. Silencing of circ-RNF111 restrained cell viability, colony formation, migration, invasion, cell cycle process and glycolysis and induced apoptosis in GC cells in vitro. Circ-RNF111 positively regulated KLF12 expression via absorbing miR-876-3p. MiR-876-3p downregulation reversed the impacts of circ-RNF111 silencing on GC cell malignant phenotypes. MiR-876-3p overexpression repressed GC cell growth, metastasis and glycolysis, inhibited apoptosis and arrested cell cycle, while KLF12 elevation weakened the effects. Besides, circ-RNF111 knockdown inhibited tumor growth in vivo. CONCLUSION: Circ-RNF111 knockdown relieved the development of GC by regulating miR-876-3p/KLF12 axis.

Circ_0093887 upregulates CCND2 and SUCNR1 to inhibit the ox-LDL-induced endothelial dysfunction in atherosclerosis by functioning as a miR-876-3p sponge.

Circular RNAs (circRNAs) are widely expressed in mammals and act as regulatory targets in the atherogenesis. The objective of this study was to research the biological role and molecular mechanism of circ_0093887 in oxidized low-density lipoprotein (ox-LDL)-induced atherosclerosis (AS) of human aortic endothelial cells (HAECs). Cell viability detection was performed by CCK-8 assay. Inflammatory molecules were examined using ELISA. Flow cytometry was used to measure cell-cycle progression and cell apoptotic rate. Caspase 3 activity was determined using caspase 3 activity assay. The expression levels of circ_0093887, miR-876-3p, CCND2 and SUCNR1 were assayed by quantitative real-time polymerase chain reaction (qRT-PCR). Dual-luciferase reporter, RNA immunoprecipitation (RIP) and RNA pull-down assays were used for the target analysis. EdU assay, wound healing assay/transwell assay and tube formation assay were, respectively, used to assess the effects of circ_0093887/miR-876-3p axis on cell proliferation, migration and tube formation. Oxidized low-density lipoprotein inhibited cell viability and cell-cycle progression but induced the inflammatory response and cell apoptosis. Circ_0093887 was downregulated and miR-876-3p was upregulated in AS patients and ox-LDL-treated HAECs. Functionally, the overexpression of circ_0093887 abrogated the cell injury of HAEC exposed to ox-LDL. For the functional mechanism, we found that circ_0093887 was a sponge for miR-876-3p and miR-876 targeted CCND2 or SUCNR1. The reverted experiment indicated that the function of circ_0093887 was achieved by sponging miR-876-3p. Meanwhile, miR-876-3p inhibitor relieved the inhibitory regulation of circ_0093887 knockdown in cell proliferation, migration and tube formation. Downregulation of miR-876-3p also alleviated the ox-LDL-induced cell injury by upregulating the expression of CCND2 or SUCNR1. Furthermore, circ_0093887 was validated to regulate the levels of CCND2 and SUCNR1 via the sponge effect on miR-876-3p. The protective effects of circ_0093887 on HAECs from ox-LDL were also ​alleviated by repressing the CCND2 and SUCNR1 levels. These findings suggested that circ_0093887 protected HAEC against the ox-LDL-induced inflammatory and apoptotic damages by targeting the miR-876-3p/CCND2 or miR-876/SUCNRA axis. Circ_0093887 could act as a potential therapeutic biomarker for AS patients.

The lncRNA SLCO4A1-AS1/miR-876-3p/RBBP6 axis regulates cell proliferation and apoptosis in acute lymphocytic leukemia via the JNK signaling pathway.

INTRODUCTION: Acute lymphocytic leukemia (ALL) is a hematologic malignancy caused by the clonal proliferation of immature lymphocytes. Long noncoding RNAs (lncRNAs) have been reported as critical regulators in several cancers, including ALL. LncRNA SLCO4A1 antisense RNA 1 (SLCO4A1-AS1) has been revealed to be implicated in tumorigenesis of several cancers. Our study focused on the role of SLCO4A1-AS1 in ALL. METHODS: RT-qPCR, Western blot analysis, CCK-8, EdU, and Flow cytometry analysis were used to explore the biological function of SLCO4A1-AS1 in ALL cellular processes. Luciferase reporter and RNA pull-down assays were applied to explore the mechanism of SLCO4A1-AS1 in ALL cells. RESULTS: SLCO4A1-AS1 was upregulated in ALL tissues and cell lines. We found that suppression of SLCO4A1-AS1 suppressed ALL cell proliferation and facilitated cell apoptosis. Our result confirmed that SLCO4A1-AS1 acted as a ceRNA by sponging microRNA 876-3p (miR-876-3p) to upregulate retinoblastoma binding protein 6 (RBBP6) expression in ALL cells. Moreover, SLCO4A1-AS1 activated the JNK signaling pathway by upregulating RBBP6. Rescue assays revealed that the activation of the JNK signaling or overexpression of RBBP6 revered the suppressive effect of SLCO4A1-AS1 knockdown on growth of ALL cells. CONCLUSION: SLCO4A1-AS1 promoted cell growth of ALL by the miR-876-3p/RBBP6 axis to activate the JNK signaling pathway.

CircTP53 promotes colorectal cancer by acting as a miR-876-3p sponge to increase cyclin-dependent kinase-like 3 expression.

According to ceRNA theory, circular RNAs could regulate certain protein expression through targeting corresponding microRNAs to affect the progression of multiple diseases, including colorectal cancer. CircTP53 (hsa_circ_0107702), highly expressed in thyroid cancer tissues, could promote the proliferation of thyroid cancer. However, the function of circTP53 in colorectal cancer is still unclear. In our study, we found circTP53 was significantly up-regulated in colorectal cancer tissues from patients and in colorectal cell lines. Next, using colorectal cell lines, we confirmed that circTP53 promoted the proliferation, migration and invasion, and reduced the apoptotic rate. Furthermore, through bioinformatics analysis and experimental confirmation, we found circTP53 functioned as the sponge of miR-876-3p, and miR-876-3p reversed the phenotype of circTP53 on the facilitation of colorectal cancer. Additionally, we found circTP53 promoted the progression of colorectal cancer by elevating the expression of CDKL3. At last, we suggested that circTP53 knockdown could inhibit colorectal cancer progression in vivo. In conclusion, circTP53 was highly expressed in colorectal cancer tissues, and promoted colorectal cancer progression via modulating miR-876-3p/CDKL3 axis.

LINC00519 regulates the proliferation, apoptosis, migration and invasion of gastric cancer cells through the miR-876-3p/HMGA1 axis / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：探讨长基因间非编码RNA 00519（long intergene non-coding RNA 00519，LINC00519）调控miR-876-3p/高迁移率家族蛋白A1（high mobility group protein A1，HMGA1）轴在胃癌HGC-27细胞的增殖、凋亡、迁移和侵袭中的作用。方法：采用qPCR检测胃癌细胞HGC-27和胃黏膜上皮细胞GES-1中LINC00519的表达水平。将HGC-27细胞按转染处理分为si-NC、si-LINC00519、si-LINC00519+anti-miR-NC和si-LINC00519+anti-miR-876-3p组，采用集落形成实验检测细胞克隆形成能力，流式细胞术检测细胞凋亡和周期分布，Transwell实验检测细胞迁移和侵袭。双荧光素酶报告实验和qPCR验证LINC00519与miR-876-3p、miR-876-3p与HMGA1之间的相互作用。结果：HGC-27细胞中LINC00519表达较GES-1细胞显著升高（P<0.05），转染siRNA后si-LINC00519组HGC-27细胞中LINC00519的表达水平较si-NC组显著降低（t=47.294，P<0.01）。与si-NC组比较，si-LINC00519组HGC-27细胞克隆数、迁移侵袭数、S期细胞比例均显著降低（均P<0.01），凋亡率、G0/G1期细胞比例均显著升高（均P<0.01）。与si-LINC00519+anti-miR-NC组比较，si-LINC00519+anti-miR-876-3p组HGC-27细胞克隆数、迁移侵袭数、S期细胞比例升高（均P<0.01），凋亡率、G0/G1期细胞比例显著降低（均P<0.01）。LINC00519能够靶向负调控miR-876-3p的表达，miR-876-3p靶向负调控HMGA1的表达。结论：敲降LINC00519能够通过调控miR-876-3p/HMGA1轴抑制胃癌HGC-27细胞的增殖、迁移和侵袭，诱导细胞凋亡。

The Involvement of the hsa_circ_0088494-miR-876-3p-CTNNB1/CCND1 Axis in Carcinogenesis and Progression of Papillary Thyroid Carcinoma.

Recently, growing studies have demonstrated that circular RNAs (circRNAs) function as critical players in multiple human tumors, including papillary thyroid carcinoma (PTC). However, the expression and underlying potential mechanism of circRNAs in PTC are still not fully elucidated. In this study, 14 candidate differentially expressed circRNAs (DECs) between normal thyroid tissues and benign thyroid tissues or PTC were first screened using the GSE93522 dataset by the GEO2R online tool. Then, the structural loop graphs of these 14 circRNAs were obtained through the CSCD database. After performing miRNA co-prediction by combination of CSCD and CRI databases, a potential circRNA-miRNA sub-network, consisting of 9 circRNAs and 21 miRNAs, was successfully constructed. Subsequently, the expression and prognostic values of these miRNAs were further determined by starBase, and two miRNAs, namely, miR-605-5p and miR-876-3p, were identified as key miRNAs in PTC. Then, their downstream target genes were predicted by the miRNet database. CTNNB1 and CCND1 were found to be two most potential targets of miR-876-3p by combination of multiple in silico analyses, including protein-protein interaction (PPI), hub gene screening, correlation analysis, and expression analysis. Conclusively, we established a key hsa_circ_0088494-miR-876-3p-CTNNB1/CCND1 axis linked to carcinogenesis and progression of PTC, which may provide promising therapeutic targets in treating PTC in the future.

Silencing LINC00504 inhibits cell proliferation, invasion as well as migration and promotes cell apoptosis in lung cancer cells via upregulating miR-876-3p.

LINC00504 acts as an oncogene and associates with unfavorable prognosis in patients with lung cancer. Silencing LINC00504 may be a promising strategy for treatment of lung cancer and its effects were firstly investigated in lung cancer cells this study. The gene expression level of miR-876-3p as well as LINC00504 were measured via PCR assay. The cell proliferation was investigated through Cell Counting Kit-8 (CCK-8) assay and colony formation assay. Flow cytometry was applied for detection of cell apoptosis. Wound healing and transwell assay were performed for measurement of cell migration and invasion respectively. The apoptosis related protein expressions were measured by western blot. Luciferase report assay was conducted for verification the target gene. LINC00504 was higher expressed in five types of lung cancer cells studied herein when compared with the control normal cells. LINC00504 knockdown exerted inhibitory effects on cell apoptosis, cell migration as well as cell invasion and promoted cell apoptosis. All the effects mentioned above were counteracted by miR-876-3p inhibitor. Silencing LINC00504 possessed anti-proliferation, repression of cell invasion as well as migration and pro-apoptosis effects via targeting up-regulation of miR-876-3p in lung cancer cells, proving the new therapeutic targets and highlighting the potential application in future diagnosis and treatment in lung cancer.

BRD8, which is negatively regulated by miR-876-3p, promotes the proliferation and apoptosis resistance of hepatocellular carcinoma cells via KAT5.

Bromodomain-containing 8 (BRD8), which belongs to the histone acetyl transferase (HAT) complex, functions as a driver in colorectal cancer. However, the role of BRD8 and its related regulatory mechanisms in hepatocellular carcinoma (HCC) remain unexplored. In this study, we found that the level of BRD8 mRNA in HCC was prominently higher than that in nontumor tissues. Furthermore, immunohistochemistry analysis indicated that BRD8 protein expression was upregulated in HCC compared to noncancerous tissues. The positive expression of BRD8 was closely associated with HBV infection, a tumor size ≥5 cm and an advanced TNM stage. Moreover, HCC patients with an elevated expression of BRD8 had an obvious poorer survival rate. Functionally, BRD8 knockdown markedly reduced the proliferation of Hep3B and Huh7 cells. Depletion of BRD8 obviously induced the apoptosis of HCC cells. Conversely, BRD8 overexpression promoted the proliferation and apoptosis resistance of Huh7 cells. Lysine acetyltransferase 5 (KAT5) expression was significantly upregulated in HCC tissues. In addition, BRD8 knockdown obviously reduced the level of KAT5 protein and the mRNA expression of KAT5-induced genes in both Hep3B and Huh7 cells. KAT5 knockdown showed similar effects as BRD8 silencing on HCC cell proliferation and apoptosis. The expression of miR-876-3p was downregulated and inversely correlated with the BRD8 mRNA level in HCC tissues. The expression of BRD8 protein in HCC cells was reduced by the overexpression of miR-876-3p and enhanced by the knockdown of miR-876-3p. A luciferase reporter assay demonstrated that BRD8 was a direct target of miR-876-3p. Notably, in HCC cells, the ectopic expression of miR-876-3p inhibited proliferation and induced apoptosis. In conclusion, BRD8, which was negatively regulated by miR-876-3p, facilitated proliferation and inhibited apoptosis in HCC cells by modulating KAT5.

Dysregulation of SPRR3/miR-876-3p Axis Contributes to Tumorigenesis in Non-Small-Cell Lung Cancer.

BACKGROUND: SPRR3, also known as esophagin, has been shown to be involved in the initiation and progression of numerous types of tumor. However, the biological function of SPRR3 that contributes to non-small-cell lung cancer (NSCLC) growth and migration is largely unknown. METHODS: The expression of SPRR3 and its association with EZH2 and miR-876-3p in NSCLC cells were determined by real-time PCR. Protein levels were measured by immunohistochemistry (IHC) and Western blot. Cell functions were studied by CCK-8, transwell assay, flow cytometry and dual-luciferase reporter assay. The effect of SPRR3 on tumor growth in vivo was evaluated in patient-derived xenograft (PDX) models. RESULTS: SPRR3 was up-regulated in most NSCLC cell lines and clinical tissues. Also, the correlation between SPRR3 expression and clinical features was significant. Functional studies confirmed that SPRR3 modulates cell proliferation, invasion and cell apoptosis in NSCLC via regulating EZH2, which is a well-known oncogene in NSCLC. Furthermore, SPRR3 was found to be a direct target of miR-876-3p that also plays a suppressor role in NSCLC. CONCLUSION: These findings indicated that miR-876-3p/SPRR3/EZH2 signaling cascade exerts important roles in the regulation of NSCLC, suggesting that this pathway can serve as a potential therapeutic target in NSCLC.