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Open Access Highly Accessed Research article

Baicalein mediates inhibition of migration and invasiveness of skin carcinoma through Ezrin in A431 cells

Bin Wu1, Ji Li2, Damao Huang3, Weiwei Wang3, Yu Chen3, Youxiang Liao1, Xiaowei Tang4, Hongfu Xie2* and Faqing Tang35*

Author Affiliations

1 Department of Dermatology, Xiangnan College, Chenzhou 423000, P.R. China

2 Department of Dermatology, Xiangya Hospital, Central South University, Changsha 410008, P.R. China

3 Department of Clinical Laboratory, Xiangya Hospital, Central South University, Changsha 410008, P.R. China

4 Metallurgical Science and Engineering, Central South University, Changsha 410008, P.R. China

5 Department of Clinical Laboratory, Zhuhai Hospital, Jinan University, Zhuhai 519000, P.R. China

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BMC Cancer 2011, 11:527  doi:10.1186/1471-2407-11-527

Published: 28 December 2011

Abstract

Background

Ezrin is highly expressed in skin cancer and promotes tumor metastasis. Ezrin serves as a promising target for anti-metastasis therapy. The aim of this study is to determine if the flavonoid bacailein inhibits the metastasis of skin cancer cells through Ezrin.

Methods

Cells from a cutaneous squamous carcinoma cell line, A431, were treated with baicalein at 0-60 μM to establish the non-cytotoxic concentration (NCC) range for baicalein. Following treatment with baicalein within this range, total Ezrin protein (both phosphorylated and unphosphorylated forms) and phosphorylated-Ezrin (phos-Ezrin) were detected by western blotting, and Ezrin RNA was detected in A431 cells using reverse transcription-polymerase chain reaction (RT-PCR). Thereafter, the motility and invasiveness of A431 cells following baicalein treatment were determined using wound-healing and Boyden chamber invasion assays. Short-interfering RNA (si-RNA) specifically targeting Ezrin was transfected into A431 cells, and a si-RNA Ezrin-A431 cell line was established by G418 selection. This stable cell line was transiently transfected with Ezrin and mutant Ezrin plasmids, and its motilityand invasiveness was subsequently determined to clarify whether bacailein inhibits these processes through Ezrin.

Results

We determined the range of NCCs for baicalein to be 2.5-40 μM in A431 cells. Baicalein displayed a dose- and time-dependent inhibition of expressions of total Ezrin and phos-Ezrin within this range NCCs. In addition, it exerted this inhibitory effect through the reduction of Ezrin RNA transcript. Baicalein also inhibited the motility and invasiveness of A431 skin carcinoma cells within the range of NCCs, in a dose- and time-dependent manner. A431 cell motility and invasiveness were inhibited by 73% and 80% respectively when cells were treated with 20 μM baicalein. However, the motility and invasiveness of A431 cells containing the Ezrin mutant were not effectively inhibited by baicalein.

Conclusions

Baicalein reduces the migration and invasiveness of A431 cells through the inhibition of Ezrin expression, which leads to the suppression of tumor metastasis.