β-arrestin2 的过表达诱导肾细胞癌细胞周期 G1 期阻滞并抑制致瘤性。

Overexpression of β-arrestin2 induces G1-phase cell cycle arrest and suppresses tumorigenicity in renal cell carcinoma.

机构信息

Department of Urology, the Third Affiliated Hospital of Soochow University, Changzhou, China.

出版信息

Eur Rev Med Pharmacol Sci. 2017 Apr;21(8):1729-1737.

Abstract

OBJECTIVE

The objective of this study was to investigate the role of β-arrestin2 in the proliferation, migration, apoptosis, cell cycle and clone formation of renal cell carcinoma (RCC) cell lines and to explore the possible mechanism of β-arrestin2 in RCC invasion and metastasis to find a new therapeutic target.

MATERIALS AND METHODS

Cell proliferation, migration, apoptosis, cell cycle and clone formation were analyzed after RCC cell lines (786-0 and CaKi) and transfected with β-arrestin2 overexpression plasmid. Using small interfering RNA (siRNA) interference technology abrogates β-arrestin2 overexpression, and changes in cell proliferation, migration, apoptosis, cell cycle and clone formation were analyzed. The expression levels of total IkBa, IkBa phosphorylation (P-IkBa) and NFkB P65 in 786-0 cells were examined after transfection with β-arrestin2 overexpression plasmid to explore the mechanism of β-arrestin2.

RESULTS

After transfection with β-arrestin2 overexpression plasmid, the abilities of proliferation, migration, and cloning formation in 786-0 and CaKi cells decreased significantly, the apoptosis rate increased significantly, and the cell cycles were blocked in the G1 phase. After siRNA reduced the expression of β-arrestin2, the abilities to proceed through cell proliferation, migration, apoptosis, the cell cycle and clone formation were enhanced. The P-IkBa level in 786-0 cells decreased significantly after transfection, while the expression of P-IkBa in the control group remained high. The expression of NFkB P65 was high in the control group and low in the transfection group.

CONCLUSIONS

The overexpression of β-arrestin2 can inhibit the growth of RCC cells in vitro, and β-arrestin2 acts as a tumor suppressor gene in RCC. The main mechanism may directly suppress the phosphorylation of IkBa and indirectly suppress NFkB activation. Thus, β-arrestin2 is expected to be an important marker of RCC prognosis and a new therapeutic target.

摘要

目的

本研究旨在探讨β-arrestin2 在肾细胞癌（RCC）细胞系增殖、迁移、凋亡、细胞周期和克隆形成中的作用，并探讨β-arrestin2 在 RCC 侵袭和转移中的可能机制，以寻找新的治疗靶点。

材料与方法

用β-arrestin2 过表达质粒转染 RCC 细胞系（786-0 和 CaKi）后，分析细胞增殖、迁移、凋亡、细胞周期和克隆形成。采用小干扰 RNA（siRNA）干扰技术阻断β-arrestin2 过表达，分析细胞增殖、迁移、凋亡、细胞周期和克隆形成的变化。转染β-arrestin2 过表达质粒后，检测 786-0 细胞总 IkBa、IkBa 磷酸化（P-IkBa）和 NFkB P65 的表达水平，探讨β-arrestin2 的作用机制。

结果

转染β-arrestin2 过表达质粒后，786-0 和 CaKi 细胞的增殖、迁移和克隆形成能力明显下降，凋亡率明显升高，细胞周期被阻滞在 G1 期。siRNA 降低β-arrestin2 表达后，细胞增殖、迁移、凋亡、细胞周期和克隆形成能力增强。转染后 786-0 细胞 P-IkBa 水平明显下降，而对照组 P-IkBa 表达仍较高。对照组 NFkB P65 表达较高，转染组表达较低。