黄芩素通过 Ezrin 介导抑制 A431 细胞皮肤癌细胞的迁移和侵袭。
Baicalein mediates inhibition of migration and invasiveness of skin carcinoma through Ezrin in A431 cells.
机构信息
Department of Dermatology, Xiangnan College, Chenzhou 423000, PR China.
出版信息
BMC Cancer. 2011 Dec 28;11:527. doi: 10.1186/1471-2407-11-527.
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 motility and 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.
背景
Ezrin 在皮肤癌中高表达，并促进肿瘤转移。Ezrin 是一种有前途的抗转移治疗靶点。本研究旨在确定黄酮类化合物白杨素是否通过 Ezrin 抑制皮肤癌细胞的转移。
方法
用白杨素处理皮肤鳞状细胞癌细胞系 A431，浓度范围为 0-60μM，以确定白杨素的非细胞毒性浓度（NCC）范围。在该范围内用白杨素处理后，通过 Western blot 检测总 Ezrin 蛋白（磷酸化和非磷酸化形式）和磷酸化 Ezrin（phos-Ezrin），并用逆转录-聚合酶链反应（RT-PCR）检测 A431 细胞中的 Ezrin RNA。然后，通过划痕愈合和 Boyden 室侵袭试验测定白杨素处理后 A431 细胞的迁移和侵袭能力。用特异性靶向 Ezrin 的短发夹 RNA（si-RNA）转染 A431 细胞，并用 G418 选择建立 Ezrin-A431 稳定细胞系。该稳定细胞系瞬时转染 Ezrin 和突变型 Ezrin 质粒，随后测定其迁移和侵袭能力，以明确白杨素是否通过 Ezrin 抑制这些过程。
结果
我们确定了白杨素在 A431 细胞中的 NCC 范围为 2.5-40μM。在这个 NCC 范围内，白杨素表现出剂量和时间依赖性的总 Ezrin 和 phos-Ezrin 表达抑制。此外，它通过减少 Ezrin RNA 转录来发挥这种抑制作用。白杨素还以剂量和时间依赖性方式抑制 A431 皮肤癌细胞的迁移和侵袭。当用 20μM 白杨素处理细胞时，A431 细胞的迁移和侵袭分别抑制了 73%和 80%。然而，含有 Ezrin 突变体的 A431 细胞的迁移和侵袭不能被白杨素有效抑制。
结论
白杨素通过抑制 Ezrin 表达减少 A431 细胞的迁移和侵袭，从而抑制肿瘤转移。
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