Overexpression Of Mir-30c-5p Reduces Cellular Cytotoxicity And Inhibits The Formation Of Kidney Stones Through Atg5

MicroRNAs (miRNAs or miRs) are critical regulators in various diseases. In the current study, the role of miR-30c-5p in the formation of sodium oxalate-induced kidney stones was investigated. For this purpose, human renal tubular epithelial cells (HK-2 cells) were incubated with sodium oxalate at the concentrations of 100, 250, 500, 750 and 1,000 mu M. Cell viability and the miR-30c-5p expression level were respectively measured by CCK-8 assay and RT-qPCR. After separately transfecting miR-30c-5p mimic and inhibitor into the HK-2 cells, the cell apoptotic rate, the levels of mitochondrial membrane potential (MMP) and ROS were determined by flow cytometry. The levels of oxidative stress indicators [lactate dehydrogenase (LDH), malondialdehyde (MDA), superoxide dismutase (SOD) and catalase (CAT)] were determined using commercial kits. Crystal-cell adhesion assay was performed to evaluate the crystal adhesion capacity in vitro. miR-30c-5p binding at autophagy related 5 (ATG5) was predicted by TargetScan7.2 and further verified by dual-luciferase reporter assay. Rescue experiments were performed to confirm the molecular mechanisms underlying sodium oxalate-induced kidney formation in HK-2 cells. The results revealed that sodium oxalate decreased the viability of HK-2 cells in a concentration-dependent manner, and that miR-30c-5p expression was significantly downregulated by exposure to 750 mu M sodium oxalate. In addition, the increase in cell apoptosis and crystal number, and the upregulated levels of LDH, MDA and ROS were reversed by the overexpression of miR-30c-5p. Moreover, the overexpression of miR-30c-5p upregulated the levels of SOD, CAT and MMP induced by sodium oxalate. ATG5 was directly regulated by miR-30c-5p, and the inhibition of cell cytotoxicity and crystal-cell adhesion induced by miR-30c-5p mimic was blocked by ATG5. These data indicated that the overexpression of miR-30c-5p alleviated cell cytotoxicity and crystal-cell adhesion induced by sodium oxalate through ATG5. Thus, the current study provides a better understanding of the role of miR-30c-5p in sodium oxalate-induced kidney stones.