Institute of Nephrology, State Key Laboratory of Kidney Disease, Chinese People's Liberation Army General Hospital, Beijing 100853, China.
Chin Med J (Engl). 2012 Feb;125(3):523-6.
Cisplatin (DDP) is one of most effective and most commonly used therapeutic agent in treating tumors, it can accumulate in the kidney and lead to acute renal failure. MicroRNA-181a can induce cell apoptosis by suppressing the expression of Bcl-2 family. In the present study, we investigated the role of microRNA-181a in the apoptosis of tubular epithelial cell induced by DDP.
HK-2 cells were cultured, transfected with microRNA-181a inhibitor for 48 hours, and stimulated with 50 µmol/L cisplatin for 24 hours. MicroRNA-181a expression was analyzed by real time PCR, and cell apoptosis was detected by flow cytometry. Moreover, Bcl-2 and Bcl-2-associated X protein (Bax) expression were measured by Western blotting.
MicroRNA-181a expression significantly down-regulated in cells transfected with microRNA-181a inhibitor, compared with that in untransfectd cells (21.19 ± 2.01 vs. 38.87 ± 1.97, P < 0.05). Cell apoptosis induced by DDP significantly decreased in cells transfected with MicroRNA-181a inhibitor. Compared with DDP treated cells alone, Bcl-2 expression strikingly was up-regulated and Bax expression was down-regulated in cells transfected with microRNA-181a inhibitor.
One pathway of DDP induces apoptosis of tubular epithelial cell by suppressing Bcl-2 expression is achieved by regulating the target gene of MicroRNA-181a.
顺铂（DDP）是治疗肿瘤最有效、最常用的治疗药物之一，它可以在肾脏中积累，导致急性肾衰竭。miRNA-181a 可以通过抑制 Bcl-2 家族的表达诱导细胞凋亡。本研究旨在探讨 miRNA-181a 在 DDP 诱导肾小管上皮细胞凋亡中的作用。
培养 HK-2 细胞，用 miRNA-181a 抑制剂转染 48 小时，用 50µmol/L 顺铂刺激 24 小时。用实时 PCR 分析 miRNA-181a 的表达，用流式细胞术检测细胞凋亡。此外，用 Western blot 法检测 Bcl-2 和 Bcl-2 相关 X 蛋白（Bax）的表达。
与未转染细胞相比，用 miRNA-181a 抑制剂转染的细胞中 miRNA-181a 的表达明显下调（21.19±2.01 比 38.87±1.97，P<0.05）。用 miRNA-181a 抑制剂转染的细胞中 DDP 诱导的细胞凋亡明显减少。与单独用 DDP 处理的细胞相比，用 miRNA-181a 抑制剂转染的细胞中 Bcl-2 的表达明显上调，Bax 的表达明显下调。
DDP 通过抑制 Bcl-2 表达诱导肾小管上皮细胞凋亡的一条途径是通过调节 miRNA-181a 的靶基因来实现的。
