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Updated: Jun 23, 2025
Studying TGF-β Signaling and TGF-β-induced Epithelial-to-mesenchymal Transition in Breast Cancer and Normal Cells
Published on: October 27, 2020
Andrographolide targets EGFR to impede epithelial-mesenchymal transition in human breast cancer cells
Chutima Kaewpiboon1, Nawong Boonnak2, Abdul-Wahab Salae3
1Department of Biology, Faculty of Science and Digital Innovation, Thaksin University, Phatthalung 93210, Thailand.
Abstract:
Despite the primary surgical treatment for breast cancer patients, malignant invasiveness and metastasis remain threatening factors for women with breast cancer. As chemotherapy yields unsatisfactory results, it prompted us to search for effective natural agents with few side-effects. Although andrographolide (ADGL), a natural diterpenoid lactone isolated from Andrographis paniculata, presents anticancer effects, the molecular mechanism remains unknown. Initially, on comparing the expression of proteins related to epithelial-mesenchymal transition (EMT) between nonmetastatic cancer MCF7 cells and highly metastatic cancer MDA-MB-231 cells, we found that MDA-MB-231 cells exhibit higher protein levels of N-cadherin and vimentin and lower protein levels of E-cadherin when compared to MCF7 cells. Moreover, MDA-MB-231 cells also exhibited higher EGFR expression and activity, higher STAT1 activity and abundant HDAC4 expression. To elucidate whether these proteins are closely associated with EMT, EGFR, STAT1 or HDAC4, the proteins were silenced in MDA-MB-231 breast cancer cells by their specific siRNAs. We found that silencing these proteins reduced EMT, indicating an important role of EGFR, STAT1 and HDAC4 in EMT progression. When we treated MDA-MB-231 cells with ADGL as a potential therapeutic drug, we found that ADGL treatment inhibited cell migration and invasion. Furthermore, it also recovered E-cadherin expression and decreased N-cadherin and vimentin protein levels. ADGL treatment reduced EGFR expression at a lower concentration (1 μg/mL); however, STAT1 activity and HDAC4 expression was reduced by a higher concentration (5 μg/mL) of ADGL. Moreover, we observed that the combined treatment with ADGL and siRNAs against these proteins highly sensitized the MDA-MB-231 cells to apoptosis compared to that with ADGL and control siRNA. Collectively, our results suggest that ADGL targets EGFR, thereby inhibiting EMT in human breast cancer cells.
Insights
Andrographolide (ADGL) inhibits breast cancer metastasis by targeting EGFR, reducing epithelial-mesenchymal transition (EMT). This natural agent shows promise in reducing invasiveness and sensitizing cancer cells to apoptosis.
Area of Science:
- Oncology
- Molecular Biology
- Natural Products Chemistry
Background:
- Breast cancer metastasis and invasiveness remain significant challenges despite primary surgical treatment.
- Chemotherapy offers limited efficacy, necessitating the search for novel, natural therapeutic agents with fewer side effects.
- Andrographolide (ADGL), a compound from Andrographis paniculata, exhibits anticancer properties, but its mechanism of action, particularly in relation to metastasis, is not fully understood.
Purpose of the Study:
- To investigate the molecular mechanisms underlying the anti-metastatic effects of Andrographolide (ADGL) in breast cancer.
- To determine the role of epithelial-mesenchymal transition (EMT) and associated proteins (EGFR, STAT1, HDAC4) in breast cancer metastasis.
- To evaluate ADGL's potential as a therapeutic agent by assessing its impact on EMT, cell migration, invasion, and apoptosis.
Main Methods:
- Comparative analysis of EMT-related protein expression (E-cadherin, N-cadherin, vimentin) in nonmetastatic (MCF7) versus highly metastatic (MDA-MB-231) breast cancer cells.
- Silencing of key proteins (EGFR, STAT1, HDAC4) using small interfering RNAs (siRNAs) in MDA-MB-231 cells to assess their role in EMT.
- Treatment of MDA-MB-231 cells with ADGL to evaluate its effects on cell migration, invasion, protein expression, and apoptosis, including combination therapy with siRNAs.
Main Results:
- Highly metastatic MDA-MB-231 cells exhibited higher N-cadherin, vimentin, EGFR, STAT1, and HDAC4 levels, and lower E-cadherin compared to MCF7 cells, confirming their association with EMT.
- Silencing EGFR, STAT1, or HDAC4 reduced EMT in MDA-MB-231 cells, highlighting their crucial roles in metastasis.
- ADGL treatment inhibited MDA-MB-231 cell migration and invasion, restored E-cadherin, decreased N-cadherin and vimentin, and modulated EGFR, STAT1, and HDAC4 expression/activity in a dose-dependent manner. Combined ADGL and siRNA treatment significantly enhanced apoptosis.
Conclusions:
- EGFR, STAT1, and HDAC4 are critical regulators of epithelial-mesenchymal transition (EMT) and metastasis in human breast cancer cells.
- Andrographolide (ADGL) effectively inhibits breast cancer cell migration and invasion by targeting EGFR and consequently suppressing EMT.
- ADGL demonstrates potential as a therapeutic agent for breast cancer, particularly in reducing metastasis, and its efficacy can be enhanced through combination therapies.
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