微小RNA-183通过微小RNA-183-异柠檬酸脱氢酶2/细胞因子信号转导抑制因子6-缺氧诱导因子-1α反馈环调节肝癌（HCC）细胞中的多药耐药性。

MiR-183 modulates multi-drug resistance in hepatocellular cancer (HCC) cells via miR-183-IDH2/SOCS6-HIF-1α feedback loop.

作者信息

Wang X-J, Zhang D-L, Fu C, Wei B-Z, Li G-J

机构信息

Clinical Laboratory, the Central Hospital of Yishui, Linyi, Shandong, China.

出版信息

Eur Rev Med Pharmacol Sci. 2016 May;20(10):2020-7.

DOI:

PMID:27249600

Abstract

OBJECTIVE

MiR-183 acts as an oncomiR and is usually upregulated in hepatocellular cancer (HCC). This study aims to study the association between miR-183 dysregulation and multi-drug resistance (MDR). Also, how it is dysregulated in HCC cells was investigated.

MATERIALS AND METHODS

The association between miR-183 and HIF-1α in HCC cell line BEL-7402 and the multidrug-resistant variant BEL-7402/5-fluorouracil (BEL-7402/5-FU) were studied using qRT-PCR and Western blot analysis. The mediators involved in feedback regulation between miR-183 and HIF-1α were further studied. Then, the effect of the miR-183-SOCS6 axis on IC50 of BEL-7402/5-FU cells to 5-FU were investigated by MTT assay.

RESULTS

The expression of miR-183 and HIF-1α are positively correlated in BEL-7402 and BEL-7402/5-FU cells. IDH2 knockdown resulted in significantly increased HIF-1α expression in both BEL-7402 and BEL-7402/5-FU cells. Knockdown of SOCS6 had similar but stronger effect as miR-183 in promoting MRP2, P-gp, p-STAT3 and HIF-1α expression in BEL-7402 cells, while SOCS6 overexpression also showed similar but stronger effect as miR-183 inhibition in reducing MRP2, P-gp, p-STAT3 and HIF-1α levels in BEL-7402/5-FU cells. Both SOCS6 overexpression and miR-183 knockdown significantly increased the sensitivity of BEL-7402/5-FU cells to 5-FU. MiR-183 overexpression partly abrogated the effect of SOCS6 in enhancing 5-FU sensitivity.

CONCLUSIONS

Both HIF-1α-miR-183-IDH2-HIF-1α and HIF-1α-miR-183-SOCS6-p-STAT3-HIF-1α feedback loops are involved in miR-183 upregulation in HCC cells. MiR-183 can modulate MDR of HCC cells at least partly through suppressing SOCS6.

摘要

目的

miR-183作为一种癌基因miRNA，在肝细胞癌（HCC）中通常上调。本研究旨在探讨miR-183失调与多药耐药（MDR）之间的关联。此外，还研究了其在肝癌细胞中失调的机制。

材料与方法

采用qRT-PCR和蛋白质免疫印迹分析，研究肝癌细胞系BEL-7402及其多药耐药变体BEL-7402/5-氟尿嘧啶（BEL-7402/5-FU）中miR-183与HIF-1α之间的关联。进一步研究参与miR-183与HIF-1α之间反馈调节的介质。然后，通过MTT法研究miR-183-SOCS6轴对BEL-7402/5-FU细胞对5-氟尿嘧啶（5-FU）半数抑制浓度（IC50）的影响。

结果

在BEL-7402和BEL-7402/5-FU细胞中，miR-183和HIF-1α的表达呈正相关。IDH2基因敲低导致BEL-7402和BEL-7402/5-FU细胞中HIF-1α表达显著增加。在促进BEL-7402细胞中MRP2、P-糖蛋白、p-STAT3和HIF-1α表达方面，敲低SOCS6与miR-183具有相似但更强的作用，而在降低BEL-7402/5-FU细胞中MRP2、P-糖蛋白、p-STAT3和HIF-1α水平方面，过表达SOCS6与抑制miR-183具有相似但更强的作用。过表达SOCS6和敲低miR-183均显著增加BEL-7402/5-FU细胞对5-FU的敏感性。miR-183过表达部分消除了SOCS6增强5-FU敏感性的作用。