Zhang Lei, Meng Shijie, Yan Bo, Chen Jie, Zhou Li, Shan Letian, Wang Ying
School of Biological and Chemical Engineering, Zhejiang University of Science and Technology, Hangzhou, People's Republic of China.
The First Affiliated Hospital, Zhejiang Chinese Medical University, Hangzhou, People's Republic of China.
Onco Targets Ther. 2021 Feb 25;14:1291-1304. doi: 10.2147/OTT.S286350. eCollection 2021.
Theaflavin (TF) is a primary pigment of tea, exhibiting anti-proliferative, pro-apoptotic and anti-metastatic activities on cancer cell lines. However, it is unknown whether TF is effective in treating melanoma cells.
To determine the effects of TF on melanoma cells, we conducted in vitro assays of cell viability, DAPI staining, wound healing, transwell, and flow cytometry as well as in vivo experiments on B16F10-bearing mouse model. Real-time PCR (qPCR) and Western blot (WB) were conducted to explore the molecular actions of TF.
The cell viability assay showed that TF exerted inhibitory effect on B16F10 cells in a dose-dependent manner from 40 to 400 μg/mL, with IC values ranging from 223.8±7.1 to 103.7±7.0 μg/mL. Moreover, TF induced early and late apoptosis and inhibited migration/invasion of B16F10 cells in a dose-dependent manner, indicating its pro-apoptotic and anti-migrative effects. In vivo, TF significantly inhibited B16F10 tumor size in mice model from 40 to 120 mg/kg, which exerted higher effect than that of cisplatin. The molecular data showed that TF significantly up-regulated the mRNA expressions of pro-apoptotic genes (, and ), up-regulated the protein expressions of apoptosis-related p53 and JNK signaling molecules (ASK1, phosphorylated Chk1/2, cleaved caspase 3, phosphorylated JNK, c-JUN, cleaved PARP, and phosphorylated p53), and down-regulated the protein expressions of proliferation-related MEK/ERK and PI3K/AKT signaling molecules (phosphorylated MEK1/2, phosphorylated ERK1/2, phosphorylated PI3K, and phosphorylated AKT) as well as the expressions of MMP2 and MMP9.
It can be concluded that TB exhibited anti-proliferative, pro-apoptotic, anti-migrative, and tumor-inhibitory effects on melanoma cells through pleiotropic actions on the above pathways. This study provides new evidence of anti-melanoma efficacy and mechanism of TF, contributing to the development of TF-derived natural products for melanoma therapy.
茶黄素（TF）是茶叶的主要色素，对癌细胞系具有抗增殖、促凋亡和抗转移活性。然而，TF对黑色素瘤细胞是否有效尚不清楚。
为了确定TF对黑色素瘤细胞的影响，我们进行了细胞活力、DAPI染色、伤口愈合、Transwell和流式细胞术的体外实验，以及对携带B16F10的小鼠模型进行体内实验。进行实时定量聚合酶链反应（qPCR）和蛋白质免疫印迹法（WB）以探究TF的分子作用。
细胞活力测定表明，TF对B16F10细胞具有抑制作用，在40至400μg/mL范围内呈剂量依赖性，IC值范围为223.8±7.1至103.7±7.0μg/mL。此外，TF诱导早期和晚期凋亡，并以剂量依赖性方式抑制B16F10细胞的迁移/侵袭，表明其促凋亡和抗迁移作用。在体内，TF在40至120mg/kg剂量下显著抑制小鼠模型中的B16F10肿瘤大小，其效果高于顺铂。分子数据表明，TF显著上调促凋亡基因（、和）的mRNA表达，上调凋亡相关p53和JNK信号分子（ASK1、磷酸化的Chk1/2、裂解的半胱天冬酶3、磷酸化的JNK、c-JUN、裂解的PARP和磷酸化的p53）的蛋白质表达，并下调增殖相关的MEK/ERK和PI3K/AKT信号分子（磷酸化的MEK1/2、磷酸化的ERK1/2、磷酸化的PI3K和磷酸化的AKT）的蛋白质表达以及MMP2和MMP9的表达。
可以得出结论，TF通过对上述途径的多效性作用对黑色素瘤细胞表现出抗增殖、促凋亡、抗迁移和肿瘤抑制作用。本研究为TF的抗黑色素瘤疗效和机制提供了新的证据，有助于开发用于黑色素瘤治疗的TF衍生天然产物。
