miR-181a 在顺铂诱导的肾小管上皮细胞凋亡中的作用。

Role of microRNA-181a in the apoptosis of tubular epithelial cell induced by cisplatin.

机构信息

Institute of Nephrology, State Key Laboratory of Kidney Disease, Chinese People's Liberation Army General Hospital, Beijing 100853, China.

出版信息

Chin Med J (Engl). 2012 Feb;125(3):523-6.

PMID:22490414

Abstract

BACKGROUND

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.

METHODS

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.

RESULTS

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.

CONCLUSION

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.

摘要

背景

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。

方法

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。

结果

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。