金雀花碱通过 HSP90 介导的 STAT3 激活抑制黑色素瘤细胞 A375 的增殖和侵袭。

Columbamine suppresses proliferation and invasion of melanoma cell A375 via HSP90-mediated STAT3 activation.

机构信息

Department of Dermatovenerology, The First Affiliated Hospital of Gannan Medical University, Ganzhou City, China.

Department of Gastroenterology, Ganzhou People's Hospital, Ganzhou City, China.

出版信息

J Recept Signal Transduct Res. 2021 Feb;41(1):99-104. doi: 10.1080/10799893.2020.1794003. Epub 2020 Jul 16.

DOI:10.1080/10799893.2020.1794003

PMID:32669028

Abstract

PURPOSE

The goal of this study is to explore the effects of columbamine in melanoma cells and the signaling pathway involved.

METHODS

Human melanoma cell line A375 cells were used in this study. Cell proliferative ability was detected by MTT assay and clone formation assay. Cell migration and invasion were measured by wound healing assay and transwell assay, respectively. Protein expression was examined by Western blotting.

RESULTS

Columbamine reduced cell proliferative ability and the number of clone spots in A375 cells. Western blotting results demonstrated that expression of cleaved caspase 3, an activated cell death protease, was upregulated by 20 and 50 µM of columbamine. Wound healing results showed that the scratch width was wider in cell treated with 20 and 50 µM of columbamine than that in cell treated with 0 and 10 µM of columbamine. Phosphorylation of STAT3 and expression of HSP90 was also repressed by columbamine in a concentration-dependent manner. Overexpression of HSP90 attenuated the inhibition of cell proliferation, migration and invasion induced by columbamine.

CONCLUSION

Columbamine inhibited melanoma cell proliferation, migration, and invasion in A375 cells through inactivation of STAT3, which is mediated by HSP90.

摘要

目的

本研究旨在探讨苦参碱在黑素瘤细胞中的作用及其相关信号通路。

方法

本研究采用人黑素瘤细胞系 A375 细胞。通过 MTT 检测和克隆形成实验检测细胞增殖能力。通过划痕愈合实验和 Transwell 实验分别检测细胞迁移和侵袭能力。通过 Western blot 检测蛋白表达。

结果

苦参碱降低了 A375 细胞的增殖能力和克隆斑点数量。Western blot 结果表明，20 和 50 μM 苦参碱能上调活化细胞死亡蛋白酶 cleaved caspase 3 的表达。划痕愈合实验结果表明，与用 0 和 10 μM 苦参碱处理的细胞相比，用 20 和 50 μM 苦参碱处理的细胞的划痕宽度更宽。苦参碱还呈浓度依赖性地抑制 STAT3 的磷酸化和 HSP90 的表达。HSP90 的过表达减弱了苦参碱诱导的细胞增殖、迁移和侵袭的抑制作用。