MiR-4261 targeted YWHAE/CAST/GPX1 impairs the calcium regulatory function of PMCA4 in erythrocytes of myeloma

OBJECTIVE: Calcium is an important substance to maintain the normal life activities of erythrocytes, and calcium overload has a damaging effect on erythrocytes. Multiple myeloma (MM) has a strong role in secreting microRNA (miRNA), but whether the miRNA produced by MM can affect the calcium regulation function of erythrocytes needs to be further studied. MATERIALS AND METHODS: The intracellular calcium content was detected by atomic absorption spectroscopy (AAS). The oxidative stress level of cells was assessed by the determination of ROS/H2O2/GSH content of erythrocytes and the detection of antioxidant capacity. QRT PCR and Western Blot were used to determine the expression of gene and protein. The binding relationship between miR 4261 and target genes was determined by a dual luciferase gene reporter assay. Targeted drugs detect the effects of plasma membrane Ca2+ ATPase 4 (PMCA4) and calpain 1 on calcium regulation in erythrocytes. RESULTS: This study found that erythrocytes in MM had calcium overload and high levels of oxidative stress, which were associated with reduced plasma membrane Ca2+ ATPase4 (PMCA4) expression in erythrocytes. Expression of 14 3 3ε (Gene: YWHAE)/calpastatin (Gene: CAST)/glutathione peroxidase 1 (Gene: GPX1), which affected PMCA4 function, was also reduced. After database screening and in vitro studies, it was found that miR 4261 is the only miRNA in MM that can simultaneously downregulate the expression of 14 3 3ε/calpastatin/GPX1. CONCLUSION: MiR 4261 can target down regulation of 14 3 3ε/calpastatin/GPX1 to affect erythrocyte PMCA4, resulting in erythrocyte calcium overload and peroxidative damage. MiR 4261 may serve as one of the potential markers to measure erythrocyte damage in MM.