Knockdown of Chloride Channel-3 Inhibits Breast Cancer Growth In Vitro and In Vivo.
10.4048/jbc.2018.21.2.103
- Author:
Fang Min ZHOU
1
;
Yun Ying HUANG
;
Tian TIAN
;
Xiao Yan LI
;
Yong Bo TANG
Author Information
1. Department of Pharmacology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China. tangyb@mail.sysu.edu.cn
- Publication Type:Original Article
- Keywords:
Breast neoplasms;
Cell proliferation;
Chloride channel-3;
Insulin-like growth factor 1
- MeSH:
Animals;
Blotting, Western;
Breast Neoplasms*;
Breast*;
Bromodeoxyuridine;
Cell Cycle;
Cell Line;
Cell Proliferation;
Chloride Channels;
Cyclin D1;
Cyclin E;
Cyclins;
Flow Cytometry;
Heterografts;
Humans;
Immunoprecipitation;
In Vitro Techniques*;
Insulin-Like Growth Factor I;
Methods;
Mice;
Proliferating Cell Nuclear Antigen;
Protein Kinases;
Real-Time Polymerase Chain Reaction;
RNA, Messenger;
Tumor Burden;
Up-Regulation
- From:Journal of Breast Cancer
2018;21(2):103-111
- CountryRepublic of Korea
- Language:English
-
Abstract:
PURPOSE: Chloride channel-3 (ClC-3) is a member of the chloride channel family and plays a critical role in a variety of cellular activities. The aim of the present study is to explore the molecular mechanisms underlying the antitumor effect of silencing ClC-3 in breast cancer. METHODS: Human breast cancer cell lines MDA-MB-231 and MCF-7 were used in the experiments. Messenger RNA and protein expression were examined by quantitative real-time polymerase chain reaction and western blot analysis. Cell proliferation was measured by the bromodeoxyuridine method, and the cell cycle was evaluated using fluorescence-activated cell sorting. Protein interaction in cells was analyzed by co-immunoprecipitation. Tumor tissues were stained with hematoxylin-eosin and tumor burden was measured using the Metamorph software. RESULTS: Breast cancer tissues collected from patients showed an increase in ClC-3 expression. Knockdown of ClC-3 inhibited the secretion of insulin-like growth factor (IGF)-1, cell proliferation, and G1/S transition in breast cancer cells. In the mouse xenograft model of human breast carcinoma, tumor growth was significantly slower in animals injected with ClC-3-deficient cells compared with the growth of normal human breast cancer cells. In addition, silencing of ClC-3 attenuated the expression of proliferating cell nuclear antigen, Ki-67, cyclin D1, and cyclin E, as well as the activation of extracellular signal-regulated protein kinases (ERK) 1/2, both in vitro and in vivo. CONCLUSION: Together, our data suggest that upregulation of ClC-3 by IGF-1 contributes to cell proliferation and tumor growth in breast cancer, and ClC-3 deficiency suppresses cell proliferation and tumor growth via the IGF/IGF receptor/ERK pathway.
