Qin Wen, Xie Wei, Yang Xi, Xia Ning, Yang Kunling
Department of Pathology, The First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi, China (mainland).
Department of Reproductive Medicine, The First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi, China (mainland).
Med Sci Monit. 2016 Mar 12;22:818-23. doi: 10.12659/msm.897187.
Acute kidney injury (AKI) is quite common in the patients who frequently use the anticancer drug cisplatin. microRNAs (miRNAs) are powerful tools in modulating the expression of key factors in disease progression, but little is known about roles of miRNAs in AKI. This study explored the expression and function of miR-449 in cisplatin-induced AKI.
MATERIAL/METHODS: Rat renal proximal tubular cell line NRK-52E was used for cisplatin treatment and miR-449 sponge transfection. MTT assay and flow cytometry were performed to detect cell viability and apoptosis in different cell groups. Protein expression of sirtuin 1 (SIRT1), acetylated p53, and BCL-associated X protein (BAX) was detected to deduce the possible regulatory mechanism of miR-449.
Results showed that cisplatin treatment in NRK-52E cells significantly up-regulated miR-449 levels (P<0.05), inhibited cell viability (P<0.05), accelerated cell apoptosis (P<0.05), and changed SIRT1, acetylated p53, and BAX protein levels (P<0.01). However, inhibiting miR-449 by its sponge transfection in cisplatin-treated cells significantly promoted cell viability (P<0.05), suppressed cell apoptosis (P<0.05), elevated SIRT1 expression (P<0.01), and inhibited acetylated p53 and BAX protein levels (P<0.001).
These results indicate that inhibiting miR-449 allows the attenuation of cisplatin-induced injury in NRK-52E cells, suggesting that miR-449 is a potential target for treating AKI. miR-449 regulates the SIRT1/p53/BAX pathway, which may be its possible mechanism in modulating cell apoptosis of cisplatin-induced AKI. Further verification and a thorough understanding are necessary for targeting miR-449 in AKI treatment.
急性肾损伤（AKI）在频繁使用抗癌药物顺铂的患者中相当常见。微小RNA（miRNA）是调节疾病进展关键因子表达的有力工具，但关于miRNA在AKI中的作用知之甚少。本研究探讨了miR-449在顺铂诱导的AKI中的表达及功能。
材料/方法：大鼠肾近端小管细胞系NRK-52E用于顺铂处理及miR-449海绵转染。采用MTT法和流式细胞术检测不同细胞组的细胞活力及凋亡情况。检测沉默调节蛋白1（SIRT1）、乙酰化p53和BCL相关X蛋白（BAX）的蛋白表达，以推断miR-449可能的调控机制。
结果显示，顺铂处理NRK-52E细胞显著上调miR-449水平（P<0.05），抑制细胞活力（P<0.05），加速细胞凋亡（P<0.05），并改变SIRT1、乙酰化p53和BAX蛋白水平（P<0.01）。然而，在顺铂处理的细胞中通过海绵转染抑制miR-449可显著促进细胞活力（P<0.05），抑制细胞凋亡（P<0.05），提高SIRT1表达（P<0.01），并抑制乙酰化p53和BAX蛋白水平（P<0.001）。
这些结果表明，抑制miR-449可减轻顺铂诱导的NRK-52E细胞损伤，提示miR-449是治疗AKI的潜在靶点。miR-449调节SIRT1/p53/BAX通路，这可能是其调节顺铂诱导的AKI细胞凋亡的潜在机制。在AKI治疗中靶向miR-449还需要进一步验证和深入了解。
