Tanshinone IIA Inhibits Epithelial-Mesenchymal Transition in Bladder Cancer Cells via Modulation of STAT3-CCL2 Signaling

• Published in: International journal of molecular sciences 2017-07, Vol.18 (8), p.1616
• Main Authors: Huang, Sung-Ying, Chang, Shu-Fang, Liao, Kuan-Fu, Chiu, Sheng-Chun
• Format: Article
• Language: English
• Subjects: Animals; Antineoplastic Agents, Phytogenic - administration & dosage; Antineoplastic Agents, Phytogenic - pharmacology; Bladder cancer; Cancer; Cell adhesion & migration; Cell Line, Tumor; Cell Movement - drug effects; Cell proliferation; chemokine (C-C motif) ligand 2; Chemokine CCL2 - genetics; Chemokine CCL2 - metabolism; Chinese medicine; Dissection; Diterpenes, Abietane - administration & dosage; Diterpenes, Abietane - metabolism; E-cadherin; Enzymatic activity; epithelial-mesenchymal transition; Epithelial-Mesenchymal Transition - drug effects; Gelatinase B; Gene expression; Gene Expression Regulation, Neoplastic; Humans; Matrix metalloproteinases; Mesenchyme; Metastases; Metastasis; Mice; Migration inhibition; Monocyte chemoattractant protein 1; N-Cadherin; Neoplasm Invasiveness; Phosphorylation; Phosphorylation - drug effects; Polymerase chain reaction; Proteins; Regulators; Salvia miltiorrhiza; signal transducer and activator of transcription 3; Signal Transduction - drug effects; Snail protein; Stat3 protein; STAT3 Transcription Factor - genetics; STAT3 Transcription Factor - metabolism; tanshinone IIA; Transcription; Urinary bladder; Urinary Bladder Neoplasms - drug therapy; Urinary Bladder Neoplasms - genetics; Urinary Bladder Neoplasms - metabolism; Vimentin; Western blotting; Xenograft Model Antitumor Assays
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms18081616

Tanshinone IIA (Tan-IIA) is an extract from the widely used traditional Chinese medicine (TCM) Danshen ( ), and has been found to attenuate the proliferation of bladder cancer (BCa) cells (The IC were: 5637, 2.6 μg/mL; BFTC, 2 μg/mL; T24, 2.7 μg/mL, respectively.). However, the mechanism of the effect of Tan-IIA on migration inhibition of BCa cells remains unclear. This study investigates the anti-metastatic effect of Tan-IIA in human BCa cells and clarifies its molecular mechanism. Three human BCa cell lines, 5637, BFTC and T24, were used for subsequent experiments. Cell migration and invasion were evaluated by transwell assays. Real-time RT-PCR and western blotting were performed to detect epithelial-mesenchymal transition (EMT)-related gene expression. The enzymatic activity of matrix metalloproteinases (MMP) was evaluated by zymography assay. Tan-IIA inhibited the migration and invasion of human BCa cells. Tan-IIA suppressed both the protein expression and enzymatic activity of MMP-9/-2 in human BCa cells. Tan-IIA up-regulated the epithelial marker E-cadherin and down-regulated mesenchymal markers such as N-cadherin and Vimentin, along with transcription regulators such as Snail and Slug in BCa cells in a time- and dose-dependent manner. Mechanism dissection revealed that Tan-IIA-inhibited BCa cell invasion could function via suppressed chemokine (C-C motif) ligand 2 (CCL2) expression, which could be reversed by the addition of CCL2 recombinant protein. Furthermore, Tan-IIA could inhibit the phosphorylation of the signal transducer and activator of transcription 3 (STAT3) (Tyr705), which cannot be restored by the CCL2 recombinant protein addition. These data implicated that Tan-IIA might suppress EMT on BCa cells through STAT3-CCL2 signaling inhibition. Tan-IIA inhibits EMT of BCa cells via modulation of STAT3-CCL2 signaling. Our findings suggest that Tan-IIA can serve as a potential anti-metastatic agent in BCa therapy.