Mouse anti-Human ROCK2 monoclonal antibody, clone 2F23

Gastric cancer (GC) is a malignant tumor with high morbidity and mortality in the world. Circular RNA hsa_circHN1_005 (circ_HN1), also termed as hsa_circ_0045602, is reported as an oncogene in GC. However, the molecular mechanism of circ_HN1 in GC development has not been fully explored. Here, we surveyed the regulatory mechanism of circ_HN1 in GC progression. The levels of circ_HN1, miR-302b-3p, and rho-associated coiled-coil containing protein kinase 2 (ROCK2) mRNA were measured by quantitative real-time polymerase chain reaction (qRT-PCR). Cell proliferation, apoptosis, colony formation, cell cycle progresion, migration, and invasion were determined by using cell counting, flow cytometry, colony formation, or transwell assays. Protein levels were detected with Western blotting. The relationship between circ_HN1 or ROCK2 and miR-302b-3p was verified via dual luciferase reporter or RNA immunoprecipitation (RIP) assays. The role of circ_HN1 in vivo was confirmed by xenograft assay. We observed that circ_HN1 and ROCK2 were upregulated while miR-302b-3p was downregulated in GC tissues and cells. Circ_HN1 silencing slowed tumor growth in vivo and impeded cell proliferation migration, invasion, and facilitated cell apoptosis in GC cells in vitro. Circ_HN1 sponged miR-302b-3p to regulate ROCK2 expression. MiR-302b-3p inhibitor reversed circ_HN1 silencing-mediated influence on the malignant behaviors of GC cells. Furthermore, ROCK2 overexpression restored miR-302b-3p mimic-mediated impacts on cell malignant behaviors in GC cells. In conclusion, circ_HN1 exerted an oncogenic role in GC through upregulating ROCK2 via sponging miR-302b-3p, offering evidence that circ_HN1 is a potential target for GC therapy.

Long noncoding RNAs (lncRNAs) have been reported to be involved in several neurological pathogenesis conditions including cerebral ischemia. In the current study, the functions of lncRNA EFNA3 on hypoxia-injured rat adrenal pheochromocytoma (PC-12) cells and the underlying molecular mechanism were studied. The expression of lncRNA EFNA3 was silenced by short hairpin RNA transfection, after which the cells were subjected with hypoxia. Cell viability, migration, invasion, and apoptosis were, respectively, determined by trypan blue staining, Transwell assay, annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) double-staining, and Western blot analysis. The cross regulation between lncRNA EFNA3 and miR-101a, as well as between miR-101a and Rho associated coiled-coil containing protein kinase 2 (ROCK2) were detected by performing quantitative real-time polymerase chain reaction, RNA pull-down, RNA immunoprecipitation, luciferase activity assay, and Western blot analysis. Studies showed that lncRNA EFNA3 was highly expressed in response to hypoxia. Deletion of lncRNA EFNA3 significantly aggravated hypoxia-induced injury in PC-12 cells, as the impairment of cell viability, migration, and invasion, and the inducement of apoptosis. LncRNA EFNA3 worked as a sponging molecule for miR-101a and miR-101a suppression-protected PC-12 cells against hypoxia-induced injury even when lncRNA EFNA3 was silenced. ROCK2 was a target gene of miR-101a. ROCK2 overexpression exhibited neuroprotective activities. Besides, ROCK2 overexpression activated Wnt/beta-catenin pathway whereas it deactivated JAK/STAT pathway upon hypoxia. Our study suggests that deletion of lncRNA EFNA3 aggravates hypoxia-induced injury in PC-12 cells by upregulating miR-101a, which further targets ROCK2.