LncRNA-ATB在草酸钙结晶诱导的HK-2细胞损伤、增殖抑制和上皮-间质转化中的作用

Role of LncRNA-ATB in Calcium Oxalate Monohydrate-Induced Cell Injury,Proliferation Inhibition and Epithelial-Mesenchymal Transformation in HK-2 Cells

目的探讨长链非编码RNA lncRNA-ATB在草酸钙结晶性肾细胞损伤中的作用。方法以一水草酸钙(COM)结晶诱导HK-2细胞损伤为模型,应用短发夹RNA(shRNA)慢病毒构建干扰lncRNA-ATB表达的稳转细胞株;qRT-PCR检测lncRNA-ATB的表达;qRT-PCR和Western-blot检测上皮-间质转化(EMT)标志物的表达;CCK-8试剂盒检测细胞增殖;LDH试剂盒检测细胞损伤。结果COM刺激的HK-2细胞中lncRNA-ATB表达显著升高(P<0.05),细胞发生损伤,增殖受抑制且发生EMT;COM刺激后HK-2细胞的上皮细胞标志物E-cadherin和ZO-1表达水平明显降低、而间质细胞标志物N-cadherin和Vimentin表达水平显著上升(P<0.01)。干扰lncRNA-ATB的表达显著缓解COM刺激所致的细胞损伤(P<0.01)、增殖抑制(P<0.01)和EMT(P<0.05)。结论LncRNA-ATB可促进COM诱导的HK-2细胞损伤、增殖抑制和EMT,提示其可能参与调控草酸钙结晶性肾损伤。

Objective To investigate the role of lncRNA-ATB in calcium oxalate crystal-induced renal injury.Methods HK-2 cells were exposed to calcium oxalate monohydrate(COM)to establish renal injury model.The stable lncRNA-ATB knockdown cell line was constructed using the shRNA lentivirus vectors.The expression of lncRNA-ATB was detected by qRT-PCR.The expression of epithelial-mesenchymal transformation(EMT)markers was measured by qRT-PCR and Western blot.Cell proliferation and injury were determined using CCK-8 and LDH assay kits,respectively.Results COM exposure induced an increase in lncRNA-ATB expression(P<0.05)and resulted in cell injury,proliferation inhibition and EMT in HK-2 cells.Furthermore,epithelial cell markers E-cadherin and ZO-1 were down-regulated and mesenchymal markers N-cadherin and Vimentin were up-regulated in HK-2 cells exposed to COM(P<0.01).Knockdown of lncRNA-ATB significantly alleviated COM-induced cell damage,proliferation inhibition and EMT(P<0.05 or P<0.01).Conclusion LncRNA-ATB participates in the regulation of calcium oxalate crystalline kidney injury by promoting COM-induced cell injury,proliferation inhibition and EMT.